Patent Publication Number: US-2020276238-A1

Title: Chimeric cytokine receptors bearing a pd-1 ectodomain

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     The present application claims the benefit of priority to U.S. Provisional Application No. 62/812,799, filed on Mar. 1, 2019; U.S. Provisional Application No. 62/894,659, filed on Aug. 30, 2019; and U.S. Provisional Application No. 62/980,737, filed on Feb. 24, 2020, the contents of all of which are hereby incorporated by reference in their entireties. 
    
    
     SEQUENCE LISTING 
     The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Feb. 24, 2020, is named AT-024_04US_SL.txt and is 554,240 bytes in size. 
     BACKGROUND 
     Adoptive transfer of immune cells (e.g. T-cells) genetically modified to recognize malignancy-associated antigens is showing promise as a new approach to treating cancer. For example, T-cells can be genetically modified to express chimeric antigen receptors (CARs), which are fusion proteins comprised of an antigen recognition moiety and T-cell activation domains. 
     T-cell proliferation, cytotoxic potency and persistence is driven by signal transduction pathways. Conventional CAR designs provide two signals—CD3zeta activation (Signal 1) and co-stimulation (Signal 2, e.g. via 4-1BB, OX40, and/or CD28 expression). In some contexts, a third signal (Signal 3), cytokine-induced cytokine receptor signaling (e.g. cytokine support for immune potentiation), may be desirable. Approaches to provide Signal 3 have however been met with significant limitations. 
     One approach to provide cytokine support includes combining CAR-T-cell therapy with systemic infusions of recombinant cytokines/cytokine mimetics, and engineering CAR-T-cells to secrete/express cytokines extracellularly. As cytokines have pleiotropic effects and can also impact the function of other cell types, the systemic administration or production of immune-potentiating cytokines by CAR-T-cells have at least two major drawbacks: (i) these approaches can cause systemic toxicity in humans, and (ii) in the context of allogeneic CAR-T-cell therapy, these approaches may cause bystander host immune-activation that could accelerate the rejection of allogeneic CAR-T-cells, thereby compromising therapeutic efficacy. Another approach to provide cytokine support was based on introducing a constitutively activated dimerized cytokine receptor, an IL-7Ra—this limits the nature (IL-7 signaling only) and magnitude of signaling output. Yet another approach to provide cytokine support involved incorporating Signal 3 directly into the CAR molecule (Nat Med. 2018 March; 24(3):352-359.). A limitation of this approach is that the strength of Signal 3 dependent on the strength of CAR activation. In the absence of target (and CAR activation), Signal 3 would not be transduced. 
     Needed are solutions to circumvent these drawbacks by targeting cytokine signals specifically to CAR-T-cells in a context-dependent manner, thus allowing for an improved safety profile and therapeutic efficacy. Provided herein and compositions and methods that address this need. 
     SUMMARY 
     Provided herein are inducible PD-1 chimeric cytokine receptors, active, for example, when engaged with PD-1 ligands or activated with an anti-PD-1 antibody. When present on chimeric antigen receptor (CAR)-bearing immune cells, and engaged with PD-1 ligands and/or activated with an anti-PD-1 antibody, such receptors allow for increased cytokine receptor signaling (Signal 3), leading to increased immune cell activation, proliferation, persistence, and/or potency of the CAR-bearing immune cells. Accordingly, the PD-1 chimeric cytokine receptors of the disclosure allow for cytokine signals to be transmitted into the immune cell with endogenous PD-1 ligands (PD-L1, PD-L2), whereby blocking their immune-suppressive signals, and converting them into immune-potentiating signals that can work in concert with, or synergize, CAR-driven activity. Moreover, as clinically approved anti-PD-1 antibodies can cluster and activate the PD-1 chimeric cytokine receptors of the disclosures, patients treated with anti-PD-1 may benefit not only from the blockage of the endogenous PD-1 signaling, but from also the activation of cytokine signaling in cells bearing the PD-1 chimeric cytokine receptors. 
     In some embodiments, provided herein are constitutively active PD-1 chimeric cytokine receptors. When present on chimeric antigen receptor (CAR)-bearing immune cells, these constitutively active receptors allow for increased basal cytokine receptor signaling (Signal 3). In some embodiments, the activity of the constitutively active PD-1 chimeric cytokine receptors can be increased when engaged with PD-1 ligands or activated with an anti-PD-1 antibody. 
     Accordingly, in one aspect, provided herein is a PD-1 chimeric cytokine receptor comprising: (a) a PD-1 ectodomain; (b) a transmembrane domain; (c) a Janus Kinase (JAK)-binding domain; and (d) a recruiting domain. The PD-1 ectodomain may comprise a portion of the extracellular region of a PD-1 protein, or may comprise a PD-1 ligand antigen binding domain. 
     In a related aspect provided herein is a polynucleotide encoding any one of the chimeric cytokine receptors of the disclosure, and an expression vector comprising such polynucleotide. In some embodiments, the polynucleotide further encodes for a chimeric antigen receptor (CAR), wherein the CAR binds to a target of interest. The target of interest can be any molecule of interest, including, for example, without limitation any one or more of those presented in Table 11. 
     In another aspect, provided herein is an engineered immune cells comprising at least one chimeric antigen receptor (CAR) and at least one chimeric cytokine receptor of the disclosure. In some embodiments the immune cell is a T-cell. In some embodiments the immune cell is an allogeneic immune cell. In other embodiments, the immune cell is an autologous immune cell. The immune cell may be selected from the group consisting of: T-cell, dendritic cell, killer dendritic cell, mast cell, NK-cell, macrophage, monocyte, B-cell and an immune cell derived from a stem cell. In a related aspect, provided herein is a pharmaceutical composition comprising any of the engineered immune cells of the disclosure, and a kit comprising such a pharmaceutical composition. 
     In another aspect, provided herein is a method of treating a cancer in a subject, comprising administering to the subject a therapeutically effective amount of any of the engineered immune cells described herein. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1A  shows a schematic of the certain embodiments of the PD-1 chimeric cytokine receptors of the disclosure. 
         FIG. 1B  shows a schematic of the PD-1 chimeric cytokine receptor of the disclosure with a wild type PD-1 ectodomain.  FIG. 1B  also shows a schematic of the PD-1 chimeric cytokine receptor—CAR vector 
         FIG. 1C  shows a schematic of an exemplary PD-1 chimeric cytokine receptor of the disclosure. 
         FIG. 1D  shows a schematic of the PD-1 chimeric cytokine receptor—CAR vector, and the control FKBP chimeric cytokine receptor CAR vector. 
         FIGS. 2A-2D  show Stat reporter activity of the PD-1 chimeric cytokine receptor in response to PD-L1-Fc ( FIGS. 2A-B ) and anti-PD-1 ( FIGS. 2C-D ). 
         FIG. 3  shows CAR and PD-1 chimeric cytokine receptor co-expression on the Day 14 following transduction of the CAR and the PD-1 chimeric cytokine receptor. 
         FIGS. 4A-4B  show phospho-Stat5 staining in human CAR-T-cells following the treatment with PD-L1-Fc ( FIG. 4A ) or anti-PD-1 ( FIG. 4B ). 
         FIGS. 5A-5D  show a schematic of PD-1 chimeric cytokine receptors with various ectodomains engineered for enhanced avidity or affinity.  FIG. 5A  shows the prototypic WT PD-1 chimeric cytokine receptor bearing the TpoR(478-582) TM/JAK2-activating domain; 
         FIG. 5B  shows the 2×PD-1 chimeric cytokine receptor bearing two WT PD-1 ectodomains in tandem;  FIG. 5C  shows the 3×PD-1 chimeric cytokine receptor bearing three WT PD-1 ectodomains in tandem;  FIG. 5D  shows the high affinity (HA) PD-1 chimeric cytokine receptor with ectodomain mutations. 
         FIGS. 6A-6D  show the response of PD-1 chimeric cytokine receptors bearing ectodomain modifications following treatment with PD-L1-Fc ( FIGS. 6A-6B ) and anti-PD-1 ( FIGS. 6C-D ). 
         FIGS. 7A-7B  show results from the cell binding assay between PD-1 chimeric cytokine receptor CAR-T-cells and PD-L1-Fc ( FIG. 7A ) or clinically-approved anti-PD-1 antibodies ( FIG. 7B ). 
         FIGS. 8A-8B  show the response of CAR-T-cells bearing PD-1 chimeric cytokine receptor ectodomain variants following treatment with PD-L1-Fc ( FIG. 8A ) or anti-PD-1 ( FIG. 8B ). 
         FIGS. 9A-9B  show the cytotoxic response of the PD-1 chimeric cytokine receptor CAR-T-cells against target cells expressing PD-1 ligands. 
         FIGS. 10A-10B  show the amino acid sequences for the wild type TpoR and the various TM deletion ( FIG. 10 a   ) or insertion ( FIG. 10 b   ) variants. 
         FIGS. 11A-11B  show the response of a HA PD-1 chimeric cytokine receptor bearing TpoR TM deletions ( FIG. 11A ) and extensions ( FIG. 11B ) following treatment with PD-L1-Fc. 
         FIGS. 12A-12B  shows the response of HA PD-1 chimeric cytokine receptor bearing TpoR TM deletions ( FIG. 12A ) and extensions ( FIG. 12B ) following treatment with anti-PD-1 (Nivolumab). 
         FIGS. 13A-13B  show the cytotoxic response of PD-1 chimeric cytokine receptor CAR-T-cells in the absence ( FIG. 13A ) or presence ( FIG. 13B ) of anti-PD-1. 
         FIG. 14  shows the cytotoxic response of CAR-T-cells coexpressing either a DN WT PD-1, a DN HA PD-1 or the HA PD-1 chimeric cytokine receptor. 
         FIG. 15  shows a Stat reporter activity of the constitutively active, dominant negative, and inducible PD-1 chimeric cytokine receptors in response to PD-L1-Fc and crosslinked PD-L1-Fc. 
         FIG. 16  shows the cytotoxic activity of HA PD1 switch CART cells with the indicated TpoR TM domains at an E:T ratio of 1:4 and in the absence of anti-PD-1. 
         FIGS. 17A-17E  show the cytotoxic activity of HA PD1 switch CART cells with the indicated TpoR TM domains at an E:T ratio of 1:8, and in the absence or presence of anti-PD-1. 
     
    
    
     DETAILED DESCRIPTION 
     Provided herein are inducible PD-1 chimeric cytokine receptors, active when engaged with PD-1 ligands or activated with an anti-PD-1 antibody. Also provided herein are constitutively active PD-1 chimeric cytokine receptors. Provided herein are chimeric antigen receptor (CAR)-bearing immune cells (CAR-I cells, e.g. CAR-T-cells), expressing the PD-1 chimeric cytokine receptors of the disclosure. Also provided herein are methods of making and using the PD-1 chimeric cytokine receptors. 
     I. PD-1 Chimeric Cytokine Receptors 
     The inducible PD-1 chimeric cytokine receptors of the disclosure activate signaling upon, for example, binding a PD-1 ligand (e.g. PD-L1, PD-L2), or a PD-1 antibody. These receptors activate signaling when monomers of the receptor cluster and/or dimerize. In some embodiments, a monomer of the PD-1 chimeric cytokine receptor of the disclosure comprises: (a) a PD-1 ectodomain; (b) a transmembrane domain; (c) a Janus Kinase (JAK)-binding domain; and; (d) a STAT-recruiting domain (e.g. from the cytoplasmic domain of a receptor; e.g. from a cytokine receptor). In some embodiments, a monomer of the PD-1 chimeric cytokine receptor of the disclosure comprises: (a) a PD-1 ectodomain; (b) a transmembrane domain; (c) a Janus Kinase (JAK)-binding domain; and; (d) a recruiting domain (e.g. from the cytoplasmic domain of a receptor; e.g. from a cytokine receptor). The recruiting domain can be a STAT-recruiting domain, an AP1-recruiting domain, a Myc/Max-recruiting domain; or a NFkB-recruiting domain. The PD-1 chimeric cytokine receptors can bind to PD-1 ligands, and/or are clustered and activated with an anti-PD-1 antibody. The PD-1 chimeric cytokine receptors activate signaling upon for example binding a PD-L1 ligand, PD-L2 ligand, and/or a PD-1 antibody. 
     The constitutively active PD-1 chimeric cytokine receptors of the disclosure are active regardless of PD-1 ligand availability, but may increase activity in the presence of a PD-ligand. In some embodiments, a monomer of the constitutively active PD-1 chimeric cytokine receptor of the disclosure comprises: (a) a PD-1 ectodomain; (b) a transmembrane domain; (c) a Janus Kinase (JAK)-binding domain; and; (d) a STAT-recruiting domain (e.g. from the cytoplasmic domain of a receptor; e.g. from a cytokine receptor). In some embodiments, a monomer of the constitutively active PD-1 chimeric cytokine receptor of the disclosure comprises: (a) a PD-1 ectodomain; (b) a transmembrane domain; (c) a Janus Kinase (JAK)-binding domain; and; (d) a recruiting domain (e.g. from the cytoplasmic domain of a receptor; e.g. from a cytokine receptor). The recruiting domain can be a STAT-recruiting domain, an AP1-recruiting domain, a Myc/Max-recruiting domain; or a NFkB-recruiting domain. In some embodiments, constitutively active PD-1 chimeric cytokine receptors can bind to PD-1 ligands or an anti-PD-1 antibody, and/or are clustered and the activity of the receptors can be further increased by engagement with PD-1 ligands or an anti-PD-1 antibody. 
     A. PD-1 Ectodomains 
     The PD-1 chimeric cytokine receptors of the disclosure comprise a PD-1 ectodomain. The PD-1 ectodomain is the domain of the chimeric cytokine receptor that extends into the extracellular space. PD-1 ectodomains of the disclosure are capable of binding PD-1 ligands (e.g. PD-L1, PD-L2) and anti-PD-1 antibodies, leading to binding-induced signal transduction. As contemplated herein, the term “PD-1 ectodomain” comprises at least a portion of the extracellular domain of a PD-1 protein or comprises a PD-1 ligand antigen binding domain (e.g. a PD-L1 antigen binding domain, or a PD-L2 antigen binding domain). 
     In some embodiments, the PD-1 ectodomain comprises a portion of a wild type PD-1. In some embodiments, the PD-1 ectodomain comprises mutations to the wild-type PD-1. In some embodiments, the PD-1 ectodomain comprises repeats of wild-type and/or mutated PD-1 amino acid sequences. 
     In some embodiments, the PD-1 ectodomain comprises a PD-1 ligand antigen binding domain, for example, the antigen binding domain of an anti-PD-L1 antibody or an anti-PD-L2 antibody. In some embodiments, the antigen binding domain is an scFv. In some embodiments, the antigen binding domain is a single chain antibody. In some embodiments, the antigen binding domain comprises a Fab portion of a PD-1 ligand antibody. 
     Table 1 shows exemplary PD1 amino acid ectodomain sequences of the disclosure. It is noted that the expression and extracellular location of the exemplary PD1 amino acid sequences can be achieved with the use of a signal sequence. In an exemplary embodiment, a CD8 signal sequence (CD8SS) MALPVTALLLPLALLLHAARP (SEQ ID NO: 1) is utilized. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Exemplary Ectodomain Sequences 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ 
               
               
                 Ectodomain 
                 Amino acid sequence 
                 ID NO: 
               
               
                   
               
               
                 Wild type (WT) PD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                   2 
               
               
                 ectodomain (PD-1(21- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 207)) 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLV 
                   
               
               
                   
                 VGVVGGLLGSLVLLVWVLAVICSRAARGTIGAR 
                   
               
               
                   
                 RTGQ 
                   
               
               
                   
               
               
                 2x WTPD1 ectodomain 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                   3 
               
               
                 (PD-1(21-207)-PD-1(21- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 207)) 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLA 
                   
               
               
                   
                 VICSRAARGTIGARRTGQPGWFLDSPDRPWNPPT 
                   
               
               
                   
                 FSPALLVVTEGDNATFTCSFSNTSESFVLNWYRM 
                   
               
               
                   
                 SPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                   
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESL 
                   
               
               
                   
                 RAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVG 
                   
               
               
                   
                 VVGGLLGSLVLLVWVLAVICSRAARGTIGARRT 
                   
               
               
                   
                 GQ 
                   
               
               
                   
               
               
                 3x WTPD1 ectodomain 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                   4 
               
               
                 (PD-1(21-207)-PD-1(21- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 207)-PD-1(21-207)) 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLA 
                   
               
               
                   
                 VICSRAARGTIGARRTGQPGWFLDSPDRPWNPPT 
                   
               
               
                   
                 FSPALLVVTEGDNATFTCSFSNTSESFVLNWYRM 
                   
               
               
                   
                 SPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                   
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESL 
                   
               
               
                   
                 RAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVG 
                   
               
               
                   
                 VVGGLLGSLVLLVWVLAVICSRAARGTIGARRT 
                   
               
               
                   
                 GQPGWFLDSPDRPWNPPTFSPALLVVTEGDNATF 
                   
               
               
                   
                 TCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDR 
                   
               
               
                   
                 SQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSG 
                   
               
               
                   
                 TYLCGAISLAPKAQIKESLRAELRVTERRAEVPTA 
                   
               
               
                   
                 HPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                   
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                   
               
               
                 High-affinity (HA) PD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                   5 
               
               
                 ectodomain 
                 SFSNTSESFHVIWHRESPSGQTDTLAAFPEDRSQP 
                   
               
               
                   
                 GQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 VCGVISLAPKIQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLA 
                   
               
               
                   
                 VICSRAARGTIGARRTGQ 
                   
               
               
                   
               
               
                 FKBP(F36V) 
                 MGVQVETISPGDGRTFPKRGQTCVVHYTGMLED 
                   6 
               
               
                   
                 GKKVDSSRDRNKPFKFMLGKQEVIRGWEEGVA 
                   
               
               
                   
                 QMSVGQRAKLTISPDYAYGATGHPGIIPPHATLV 
                   
               
               
                   
                 FDVELLKLE 
                   
               
               
                   
               
               
                 Wildtype (WT) PD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                 132 
               
               
                 ectodomain (PD-1(21- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 170)) 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLV 
                   
               
               
                   
               
               
                 High-affinity (HA) PD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                 133 
               
               
                 ectodomain (21-170) 
                 SFSNTSESFHVIWHRESPSGQTDTLAAFPEDRSQP 
                   
               
               
                   
                 GQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 VCGVISLAPKIQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLV 
                   
               
               
                   
               
               
                 2x WTPD1 ectodomain  
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                 168 
               
               
                 (PD-1(21-170)-G-PD- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 1(21-170)) 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                   
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLVGPGWFLDSPDRPWNPPTFSP 
                   
               
               
                   
                 ALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPS 
                   
               
               
                   
                 NQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDF 
                   
               
               
                   
                 HMSVVRARRNDSGTYLCGAISLAPKAQIKESLRA 
                   
               
               
                   
                 ELRVTERRAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                   
               
               
                 3x WTPD1 ectodomain 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                 169 
               
               
                 (PD-1(21-170)-G-PD- 
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQ 
                   
               
               
                 1(21-170)-G-PD-1(21- 
                 PGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTY 
                   
               
               
                 170)) 
                 LCGAISLAPKAQIKESLRAELRVTERRAEVPTAHP 
                   
               
               
                   
                 SPSPRPAGQFQTLVGPGWFLDSPDRPWNPPTFSP 
                   
               
               
                   
                 ALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPS 
                   
               
               
                   
                 NQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDF 
                   
               
               
                   
                 HMSVVRARRNDSGTYLCGAISLAPKAQIKESLRA 
                   
               
               
                   
                 ELRVTERRAEVPTAHPSPSPRPAGQFQTLVGPGW 
                   
               
               
                   
                 FLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSN 
                   
               
               
                   
                 TSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQ 
                   
               
               
                   
                 DCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCG 
                   
               
               
                   
                 AISLAPKAQIKESLRAELRVTERRAEVPTAHPSPS 
                   
               
               
                   
                 PRPAGQFQTLV 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the PD-1 ectodomain comprises a wild type PD-1 ectodomain. 
     In some embodiments, the PD-1 ectodomain comprises a wild type PD-1 and comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the PD-1 ectodomain comprises a wild type PD-1 and comprises the amino acid sequence of SEQ ID NO: 132. 
     In some embodiments, the PD-1 ectodomain comprises mutations to the wild type PD-1 ectodomain sequence. In some embodiments, the PD-1 ectodomain sequence is a high affinity PD-1 ectodomain. In some embodiments, the PD-1 ectodomain sequence is a high affinity PD-1 ectodomain and comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the PD-1 ectodomain sequence is a high affinity PD-1 ectodomain and comprises the amino acid sequence of SEQ ID NO: 133. 
     In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of greater than one wild type PD-1 ectodomain sequence, for example at least two, at least three, at least four, or at least five wild type PD-1 ectodomain sequences. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of two wild type PD-1 ectodomain sequences and comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of two wild type PD-1 ectodomain sequences and comprises the amino acid sequence of SEQ ID NO: 168. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of three wild type PD-1 ectodomain sequences and comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of three wild type PD-1 ectodomain sequences and comprises the amino acid sequence of SEQ ID NO: 169. 
     In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of greater than one high affinity PD-1 ectodomain sequence, for example at least two, at least three, at least four, or at least five high affinity PD-1 ectodomain sequences. 
     In some embodiments, the PD-1 ectodomain comprises a high affinity PD-1 and comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the high affinity PD-1 ectodomain comprises the amino acid sequence of SEQ ID NO: 133. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of two high affinity PD-1 ectodomain sequences. In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of three high affinity PD-1 ectodomain sequences. 
     In some embodiments, the PD-1 ectodomain comprises a tandem arrangement of a combination of wild type and high affinity PD-1 ectodomain sequences, for example a combination of at least two, at least three, at least four, or at least five wild type and high affinity PD-1 ectodomain sequences. 
     In some embodiments, the greater than one wide type PD-1 ectodomain, or greater than one high affinity PD-1 ectodomain, or a combination thereof, can be connected either directly, or via a linker. In some embodiments, the linker can comprise one or more amino acid residues. In some embodiments, the linker is a Gly. 
     In some embodiments, the PD-1 ectodomain is a dominant negative. Table 2 shows exemplary PD-1 dominant negative (DN) sequences of the disclosure. The DN sequences of Table 2 may be expressed with the aid of a signal sequence, e.g. a CD8SS signal sequence of SEQ ID NO: 1 
     
       
         
           
               
             
               
                 TABLE 2  
               
             
            
               
                   
               
               
                 PD1 dominant negative (DN) sequences: 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ 
               
               
                 PD1 DN 
                 Amino acid sequence 
                 ID NO: 
               
               
                   
               
               
                 Dominant 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 7 
               
               
                 negative wild 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 type PD1 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 (21-207) 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                   
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                   
               
               
                 Dominant 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 8 
               
               
                 negative high 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 affinity PD1 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                   
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                   
                 AVICSRAARGTIGARRTGQ 
               
               
                   
               
            
           
         
       
     
     B. Transmembrane Domains 
     The PD-1 chimeric cytokine receptors of the disclosure comprise transmembrane domains. Such transmembrane domains are coupled to the extracellular PD-1 ectodomain on the N-terminus, and to additional intracellular/cytoplasmic domains on the C-terminus. In some embodiments, the coupling is achieved optionally through a linker. In some embodiments, the linker can comprise one or more amino acid residues. 
     As used herein, the transmembrane domains are capable of insertion into the membrane of a cell in which it is expressed. In some embodiments, the transmembrane domains of the disclosure span a cellular membrane, and comprise an extracellular portion, and/or an intracellular portion. 
     In some embodiments, the transmembrane domains of the disclosure are engineered and do not resemble any naturally occurring transmembrane domain, e.g. they are non-naturally occurring. 
     In other embodiments, the transmembrane domains of the disclosure are derived from naturally occurring receptors. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure are derived from, for example, one or more of the following receptors: erythropoietin receptor (EpoR), Interleukin 6 signal transducer (GP130 or IL6ST), prolactin receptor (PrlR), growth hormone receptor (GHR), granulocyte colony-stimulating factor receptor (GCSFR), and thrombopoietin receptor/myeloproliferative leukemia protein receptor (TPOR/MPLR). When derived from naturally occurring receptors, the entire receptor, or the entire transmembrane sequence of the receptor may not be necessary to effectuate constitutive activation and constitutive JAK binding/activation on the intracellular portion. Accordingly fragments of naturally occurring receptors may be utilized. Furthermore, certain mutations may be introduced into the transmembrane domains derived from naturally occurring receptors, to further tune the downstream JAK-dependent signaling. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring EpoR receptor. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring GP130 receptor. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring PrlR receptor. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring GHR receptor. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring GCSF receptor. 
     In some embodiments, the transmembrane and/or JAK domains of the disclosure is derived from the naturally occurring TPOR receptor. 
     Table 3 provides exemplary full length sequences of naturally occurring receptors provided in the disclosure, from which the transmembrane and/or JAK domains are derived. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Exemplary Naturally Occurring Receptors 
               
            
           
           
               
               
            
               
                   
                 SEQ ID 
               
               
                 Naturally Occurring Receptor Name 
                 NO: 
               
               
                   
               
               
                 &gt;AAI12154.1 Erythropoietin receptor sapiens 
                  9 
               
               
                 MDHLGASLWPQVGSLCLLLAGAAWAPPPNLPDPKFESKAALLAARGPEELLCFTERLEDLVCFWEEAASA 
                   
               
               
                 GVGPGNYSFSYQLEDEPWKLCRLHQAPTARGAVRFWCSLPTADTSSFVPLELRVTAASGAPRYHRVIHIN 
                   
               
               
                 EVVLLDAPVGLVARLADESGHVVLRWLPPPETPMTSHIRYEVDVSAGNGAGSVQRVEILEGRTECVLSNL 
                   
               
               
                 RGRTRYTFAVRARMAEPSFGGFWSAWSEPVSLLTPSDLDPLILTLSLILVVILVLLTVLALLSHRRALKQ 
                   
               
               
                 KIWPGIPSPESEFEGLFTTHKGNFQLWLYQNDGCLWWSPCTPFTEDPPASLEVLSERCWGTMQAVEPGTD 
                   
               
               
                 DEGPLLEPVGSEHAQDTYLVLDKWLLPRNPPSEDLPGPGGSVDIVAMDEGSEASSCSSALASKPSPEGAS 
                   
               
               
                 AASFEYTILDPSSQLLRPWTLCPELPPTPPHLKYLYLVVSDSGISTDYSSGDSQGAQGGLSDGPYSNPYE 
                   
               
               
                 NSLIPAAEPLPPSYVACS 
                   
               
               
                   
               
               
                 &gt;AA117403.1 Interleukin 6 signal transducer (GP130, oncostatin M receptor) 
                 10 
               
               
                 [ Homo   sapiens ] 
                   
               
               
                 MLTLQTWLVQALFIFLTTESTGELLDPCGYISPESPVVQLHSNFTAVCVLKEKCMDYFHVNANYIVWKTN 
                   
               
               
                 HFTIPKEQYTIINRTASSVTFTDIASLNIQLTCNILTFGQLEQNVYGITIISGLPPEKPKNLSCIVNEGK 
                   
               
               
                 KMRCEWDRGRETHLETNFTLKSEWATHKFADCKAKRDTPTSCTVDYSTVYFVNIEVWVEAENALGKVTSD 
                   
               
               
                 HINFDPVYKVKPNPPHNLSVINSEELSSILKLTWTNPSIKSVIILKYNIQYRTKDASTWSQIPPEDTAST 
                   
               
               
                 RSSFTVQDLKPFTEYVFRIRCMKEDGKGYWSDWSEEASGITYEDRPSKAPSFWYKIDPSHTQGYRTVQLV 
                   
               
               
                 WKTLPPFEANGKILDYEVTLTRWKSHLQNYTVNATKLTVNLTNDRYVATLTVRNLVGKSDAAVLTIPACD 
                   
               
               
                 FQATHPVMDLKAFPKDNMLWVEWTTPRESVKKYILEWCVLSDKAPCITDWQQEDGTVHRTYLRGNLAESK 
                   
               
               
                 CYLITVTPVYADGPGSPESIKAYLKQAPPSKGPTVRTKKVGKNEAVLEWDQLPVDVQNGFIRNYTIFYRT 
                   
               
               
                 IIGNETAVNVDSSHTEYTLSSLTSDTLYMVRMAAYTDEGGKDGPEFTFTTPKFAQGEIEAIVVPVCLAFL 
                   
               
               
                 LTTLLGVLFCFNKRDLIKKHIWPNVPDPSKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVSVVEIEAND 
                   
               
               
                 KKPFPEDLKSLDLFKKEKINTEGHSSGIGGSSCMSSSRPSISSSDENESSQNTSSTVQYSTVVHSGYRHQ 
                   
               
               
                 VPSVQVFSRSESTQPLLDSEERPEDLQLVDHVDGGDGILPRQQYFKQNCSQHESSPDISHFERSKQVSSV 
                   
               
               
                 NEEDFVRLKQQISDHISQSCGSGQMKMFQEVSAADAFGPGTEGQVERFETVGMEAATDEGMPKSYLPQT 
                   
               
               
                 V 
                   
               
               
                 RQGGYMPQ 
                   
               
               
                   
               
               
                 &gt;XP_011512371.1 prolactin receptor isoform X2 [ Homo   sapiens ] 
                 11 
               
               
                 MKENVASATVFTLLLFLNTCLLNGQLPPGKPEIFKCRSPNKETFTCWWRPGTDGGLPTNYSLTYHREGET 
                   
               
               
                 LMHECPDYITGGPNSCHFGKQYTSMWRTYIMMVNATNQMGSSFSDELYVDVTYIVQPDPPLELAVEVKQP 
                   
               
               
                 EDRKPYLWIKWSPPTLIDLKTGWFTLLYEIRLKPEKAAEWEIHFAGQQTEFKILSLHPGQKYLVQVRCKP 
                   
               
               
                 DHGYWSAWSPATFIQIPSDFTMNDTTVWISVAVLSAVICLIIVWAVALKGYSMVTCIFPPVPGPKIKGFD 
                   
               
               
                 AHLLEKGKSEELLSALGCQDFPPTSDYEDLLVEYLEVDDSEDQHLMSVHSKEHPSQGMKPTYLDPDTDSG 
                   
               
               
                 RGSCDSPSLLSEKCEEPQANPSTFYDPEVIEKPENPETTHTWDPQCISMEGKIPYFHAGGSKCSTWPLPQ 
                   
               
               
                 PSQHNPRSSYHNITDVCELAVGPAGAPATLLNEAGKDALKSSQTIKSREEGKATQQREVESFHSETDQDT 
                   
               
               
                 PWLLPQEKTPFGSAKPLDYVEIHKVNKDGALSLLPKQRENSGKPKKPGTPENNKEYAKVSGVMDNNILVL 
                   
               
               
                 VPDPHAKNVACFEESAKEAPPSLEQNQAEKALANFTATSSKCRLQLGGLDYLDPACFTHSFH 
                   
               
               
                   
               
               
                 &gt;NP_000154.1 growth hormone receptor isoform 1 precursor [ Homo   sapiens ] 
                 12 
               
               
                 MDLWQLLLTLALAGSSDAFSGSEATAAILSRAPWSLQSVNPGLKTNSSKEPKFTKCRSPERETFSCHWTD 
                   
               
               
                 EVHHGTKNLGPIQLFYTRRNTQEWTQEWKECPDYVSAGENSCYFNSSFTSIWIPYCIKLTSNGGTVDEKC 
                   
               
               
                 FSVDEIVQPDPPIALNWTLLNVSLTGIHADIQVRWEAPRNADIQKGWMVLEYELQYKEVNETKWKMMDPI 
                   
               
               
                 LTTSVPVYSLKVDKEYEVRVRSKQRNSGNYGEFSEVLYVTLPQMSQFTCEEDFYFPWLLIIIFGIFGLTV 
                   
               
               
                 MLFVFLFSKQQRIKMLILPPVPVPKIKGIDPDLLKEGKLEEVNTILAIHDSYKPEFHSDDSWVEFIELDI 
                   
               
               
                 DEPDEKTEESDTDRLLSSDHEKSHSNLGVKDGDSGRTSCCEPDILETDFNANDIHEGTSEVAQPQRLKGE 
                   
               
               
                 ADLLCLDQKNQNNSPYHDACPATQQPSVIQAEKNKPQPLPTEGAESTHQAAHIQLSNPSSLSNIDFYAQV 
                   
               
               
                 SDITPAGSVVLSPGQKNKAGMSQCDMHPEMVSLCQENFLMDNAYFCEADAKKCIPVAPHIKVESHIQPSL 
                   
               
               
                 NQEDIYITTESLTTAAGRPGTGEHVPGSEMPVPDYTSIHIVQSPQGLILNATALPLPDKEFLSSCGYVST 
                   
               
               
                 DQLNKIMP 
                   
               
               
                   
               
               
                 &gt;XP_016855859.1 granulocyte colony-stimulating factor receptor isoform X1 
                 13 
               
               
                 [ Homo   sapiens ] 
                   
               
               
                 MARLGNCSLTWAALIILLLPGSLEECGHISVSAPIVHLGDPITASCIIKQNCSHLDPEPQILWRLGAELQ 
                   
               
               
                 PGGRQQRLSDGTQESIITLPHLNHTQAFLSCCLNWGNSLQILDQVELRAGYPPAIPHNLSCLMNLTTSSL 
                   
               
               
                 ICQWEPGPETHLPTSFTLKSFKSRGNCQTQGDSILDCVPKDGQSHCCIPRKHLLLYQNMGIWVQAENALG 
                   
               
               
                 TSMSPQLCLDPMDVVKLEPPMLRTMDPSPEAAPPQAGCLQLCWEPWQPGLHINQKCELRHKPQRGEASWA 
                   
               
               
                 LVGPLPLEALQYELCGLLPATAYTLQIRCIRWPLPGHWSDWSPSLELRTTERAPTVRLDTWWRQRQLDPR 
                   
               
               
                 TVQLFWKPVPLEEDSGRIQGYVVSWRPSGQAGAILPLCNTTELSCTFHLPSEAQEVALVAYNSAGTSRPT 
                   
               
               
                 PVVFSESRGPALTRLHAMARDPHSLWVGWEPPNPWPQGYVIEWGLGPPSASNSNKTWRMEQNGRATGFLL 
                   
               
               
                 KENIRPFQLYEIIVTPLYQDTMGPSQHVYAYSQEMAPSHAPELHLKHIGKTWAQLEWVPEPPELGKSPLT 
                   
               
               
                 HYTIFWTNAQNQSFSAILNASSRGFVLHGLEPASLYHIHLMAASQAGATNSTVLTLMTLTPEGSELHIIL 
                   
               
               
                 GLFGLLLLLTCLCGTAWLCCSPNRKNPLWPSVPDPAHSSLGSWVPTIMEELPGPRQGQWLGQTSEMSRAL 
                   
               
               
                 TPHPCVQDAFQLPGLGTPPITKLTVLEEDEKKPVPWESHNSSETCGLPTLVQTYVLQGDPRAVSTQPQSQ 
                   
               
               
                 SGTSDQVLYGQLLGSPTSPGPGHYLRCDSTQPLLAGLTPSPKSYENLWFQASPLGTLVTPAPSQEDDCVF 
                   
               
               
                 GPLLNFPLLQGIRVHGMEALGSF 
                   
               
               
                   
               
               
                 &gt;NP_005364.1 thrombopoietin receptor precursor [ Homo   sapiens ] 
                 131 
               
               
                 MPSWALFMVTSCLLLAPQNLAQVSSQDVSLLASDSEPLKCFSRTFEDLTCFWDEEEAAPSGTYQLLYAYP 
                   
               
               
                 REKPRACPLSSQSMPHFGTRYVCQFPDQEEVRLFFPLHLWVKNVFLNQTRTQRVLFVDSVGLPAPPSIIK 
                   
               
               
                 AMGGSQPGELQISWEEPAPEISDFLRYELRYGPRDPKNSTGPTVIQLIATETCCPALQRPHSASALDQSP 
                   
               
               
                 CAQPTMPWQDGPKQTSPSREASALTAEGGSCLISGLQPGNSYWLQLRSEPDGISLGGSWGSWSLPVTVDL 
                   
               
               
                 PGDAVALGLQCFTLDLKNVTCQWQQQDHASSQGFFYHSRARCCPRDRYPIWENCEEEEKTNPGLQTPQFS 
                   
               
               
                 RCHFKSRNDSIIHILVEVTTAPGTVHSYLGSPFWIHQAVRLPTPNLHWREISSGHLELEWQHPSSWAAQE 
                   
               
               
                 TCYQLRYTGEGHQDWKVLEPPLGARGGTLELRPRSRYRLQLRARLNGPTYQGPWSSWSDPTRVETATETA 
                   
               
               
                 WISLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                 EEVEPSLLEILPKSSERTPLPLCSSQAQMDYRRLQPSCLGTMPLSVCPPMAESGSCCTTHIANHSYLPLS 
                   
               
               
                 YWQQP 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the transmembrane domain of the disclosure is derived from a truncated, or otherwise modified version of the naturally occurring TPOR/MPLR receptor shown in Table 3. 
       FIGS. 10A-B  show the amino acid sequences for the wild type TpoR and the various transmembrane deletion ( FIG. 10A ) or insertion ( FIG. 10B ) variants. 
     Table 4 shows exemplary transmembrane amino acid sequences, coupled to intracellular JAK2 binding domain sequences. 
     In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 15. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 16. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 17. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 28. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 29. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 30. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 31. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 32. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 33. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 34. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 35. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 36. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 38. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 40. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 41. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 44. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 134. In some embodiments, the transmembrane domain of the PD-1 chimeric cytokine receptor comprises the amino acid sequence of SEQ ID NO: 135. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises an amino acid sequence of the transmembrane+JAK2 binding domain that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the amino acid sequence of any one of the sequences shown in Table 4. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Exemplary Transmembrane + JAK2 Binding Domain Sequences 
               
            
           
           
               
               
               
            
               
                 Transmembrane and 
                   
                 SEQ 
               
               
                 JAK2 binding domain 
                 Amino acid sequence 
                 ID NO: 
               
               
                   
               
               
                 GCSFR(614-710) 
                 LTLMTLTPEGSELHIILGLFGLLLLLTCLCGTAWL 
                 14 
               
               
                   
                 CCSPNRKNPLWPSVPDPAHSSLGSWVPTIMEEDA 
                   
               
               
                   
                 FQLPGLGTPPITKLTVLEEDEKKPVPWE 
                   
               
               
                   
               
               
                 GP130(609-700) 
                 TTPKFAQGEIEAIVVPVCLAFLLTTLLGVLFCFNK 
                 15 
               
               
                   
                 RDLIKKHIWPNVPDPSKSHIAQWSPHTPPRHNFN 
                   
               
               
                   
                 SKDQMYSDGNFTDVSVVEIEAND 
                   
               
               
                   
               
               
                 TPOR/MPLR(478-582) 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLL 
                 16 
               
               
                   
                 LLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                   
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPL 
                   
               
               
                   
                 PL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-1) 
                 SDPTRVETATETWISLVTALHLVLGLSAVLGLLL 
                 17 
               
               
                   
                 LRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRD 
                   
               
               
                   
                 TAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLP 
                   
               
               
                   
                 L 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-2) 
                 SDPTRVETATETISLVTALHLVLGLSAVLGLLLLR 
                 18 
               
               
                   
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                   
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-2+1) 
                 SDPTRVETATETLISLVTALHLVLGLSAVLGLLLL 
                 19 
               
               
                   
                 RWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                   
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-3) 
                 SDPTRVETATETSLVTALHLVLGLSAVLGLLLLR 
                 20 
               
               
                   
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                   
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-4) 
                 SDPTRVETATETLVTALHLVLGLSAVLGLLLLRW 
                 21 
               
               
                   
                 QFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAA 
                   
               
               
                   
                 LSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-4+1) 
                 SDPTRVETATETILVTALHLVLGLSAVLGLLLLR 
                 22 
               
               
                   
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                   
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-5) 
                 SDPTRVETATETVTALHLVLGLSAVLGLLLLRW 
                 23 
               
               
                   
                 QFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAA 
                   
               
               
                   
                 LSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-6) 
                 SDPTRVETATETTALHLVLGLSAVLGLLLLRWQF 
                 24 
               
               
                   
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALS 
                   
               
               
                   
                 PPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-7) 
                 SDPTRVETATETALHLVLGLSAVLGLLLLRWQFP 
                 25 
               
               
                   
                 AHYRRLRHALWPSLPDLHRVLGQYLRDTAALSP 
                   
               
               
                   
                 PKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-8) 
                 SDPTRVETATETLHLVLGLSAVLGLLLLRWQFPA 
                 26 
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPP 
                   
               
               
                   
                 KATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-9) 
                 SDPTRVETATETHLVLGLSAVLGLLLLRWQFPAH 
                 27 
               
               
                   
                 YRRLRHALWPSLPDLHRVLGQYLRDTAALSPPK 
                   
               
               
                   
                 ATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-10) 
                 SDPTRVETATETLVLGLSAVLGLLLLRWQFPAHY 
                 28 
               
               
                   
                 RRLRHALWPSLPDLHRVLGQYLRDTAALSPPKA 
                   
               
               
                   
                 TVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-11) 
                 SDPTRVETATETVLGLSAVLGLLLLRWQFPAHYR 
                 29 
               
               
                   
                 RLRHALWPSLPDLHRVLGQYLRDTAALSPPKAT 
                   
               
               
                   
                 VSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-12) 
                 SDPTRVETATETLGLSAVLGLLLLRWQFPAHYRR 
                 30 
               
               
                   
                 LRHALWPSLPDLHRVLGQYLRDTAALSPPKATV 
                   
               
               
                   
                 SDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-13) 
                 SDPTRVETATETGLSAVLGLLLLRWQFPAHYRRL 
                 31 
               
               
                   
                 RHALWPSLPDLHRVLGQYLRDTAALSPPKATVS 
                   
               
               
                   
                 DTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-14) 
                 SDPTRVETATETLSAVLGLLLLRWQFPAHYRRLR 
                 32 
               
               
                   
                 HALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                   
                 TCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-15) 
                 SDPTRVETATETSAVLGLLLLRWQFPAHYRRLRH 
                 33 
               
               
                   
                 ALWPSLPDLHRVLGQYLRDTAALSPPKATVSDT 
                   
               
               
                   
                 CEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-16) 
                 SDPTRVETATETAVLGLLLLRWQFPAHYRRLRH 
                 34 
               
               
                   
                 ALWPSLPDLHRVLGQYLRDTAALSPPKATVSDT 
                   
               
               
                   
                 CEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-17) 
                 SDPTRVETATETVLGLLLLRWQFPAHYRRLRHA 
                 35 
               
               
                   
                 LWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCE 
                   
               
               
                   
                 EVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N-18) 
                 SDPTRVETATETLGLLLLRWQFPAHYRRLRHAL 
                 36 
               
               
                   
                 WPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEE 
                   
               
               
                   
                 VEPSLLEILPKSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+1) 
                 SDPTRVETATETAWLISLVTALHLVLGLSAVLGL 
                 37 
               
               
                   
                 LLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYL 
                   
               
               
                   
                 RDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTP 
                   
               
               
                   
                 LPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+2) 
                 SDPTRVETATETAWVLISLVTALHLVLGLSAVLG 
                 38 
               
               
                   
                 LLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQY 
                   
               
               
                   
                 LRDTAALSPPKATVSDTCEEVEPSLLEILPKSSER 
                   
               
               
                   
                 TPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+3) 
                 SDPTRVETATETAWLVLISLVTALHLVLGLSAVL 
                 39 
               
               
                   
                 GLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQ 
                   
               
               
                   
                 YLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+4) 
                 SDPTRVETATETAWILVLISLVTALHLVLGLSAVL 
                 40 
               
               
                   
                 GLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQ 
                   
               
               
                   
                 YLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+5) 
                 SDPTRVETATETAWLILVLISLVTALHLVLGLSAV 
                 41 
               
               
                   
                 LGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLG 
                   
               
               
                   
                 QYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSS 
                   
               
               
                   
                 ERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+6) 
                 SDPTRVETATETAWLLILVLISLVTALHLVLGLSA 
                 42 
               
               
                   
                 VLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVL 
                   
               
               
                   
                 GQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKS 
                   
               
               
                   
                 SERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+7) 
                 SDPTRVETATETAWVLLILVLISLVTALHLVLGLS 
                 43 
               
               
                   
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                   
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPK 
                   
               
               
                   
                 SSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(N+8) 
                 SDPTRVETATETAWLVLLILVLISLVTALHLVLGL 
                 44 
               
               
                   
                 SAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHR 
                   
               
               
                   
                 VLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILP 
                   
               
               
                   
                 KSSERTPLPL 
                   
               
               
                   
               
               
                 TPOR/MPLR(478-582, 
                 SDPTRVETATETAWISLVTAL   L   LVLGL   N   AVLGLL 
                 134 
               
               
                 H499L, S505N, 
                 LLR   K   QFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                 W515K) 
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPL 
                   
               
               
                   
                 PL 
                   
               
               
                   
               
               
                 TPOR/MPLR(478-582, 
                 SDPTRVETATETAWISLVTALHLVLGL   N   AVLGLL 
                 135 
               
               
                 S505N, W515K) 
                 LLR   K   QFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                   
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPL 
                   
               
               
                   
                 PL 
               
               
                   
               
            
           
         
       
     
     C. Janus Kinase (JAK)-Binding Domains 
     The PD-1 chimeric cytokine receptors of the disclosure comprise intracellular JAK-binding domains. The JAK-binding domain is coupled to the C-terminus of the transmembrane domain, either directly, or via a linker. In some embodiments, the linker can comprise one or more amino acid residues. The JAK-binding domain is coupled to the transmembrane domain on the intracellular side of the chimeric cytokine receptor. 
     In some embodiments, the JAK-binding domain is a JAK-1-binding domain, a JAK-2 binding domain, a JAK-3 binding domain, or a TYK2 binding domain. 
     In some embodiments, the JAK-binding domains of the PD-1 chimeric cytokine receptors of the disclosure are naturally occurring, and derived from a naturally occurring receptor. 
     In some embodiments, the JAK-binding domains of the PD-1 chimeric cytokine receptors of the disclosure are synthetic. 
     Table 4 provides exemplary amino acid sequences for Transmembrane and JAK2 Binding domains of the disclosure. 
     D. Recruiting Domains 
     The PD-1 chimeric cytokine receptors of the disclosure comprise cytoplasmic domains comprising recruiting domains. The recruiting domain can be a STAT-recruiting domain, an AP1-recruiting domain, a Myc/Max-recruiting domain; or an NFkB-recruiting domain. In some embodiments, the recruiting domain is a Signal Transducer and Activator of Transcription (STAT)--recruiting (Stat-activating) domains from receptor tails (cytotails) or from cytokine receptor tails. These intracellular recruiting domains of the PD-1 chimeric cytokine receptors of the disclosure allow for the propagation of Signal 3 in an immune cell comprising a CAR and a chimeric cytokine receptor (e.g. a CAR-T-cell with a chimeric cytokine receptor of the disclosure). Cytokine signaling propagated through the Stat-recruiting domain allows for the cytokine-based immune potentiation of the cell. In some embodiments, the immune-potentiation is homeostatic, e.g. signaling gives rise to increase in immune cells bearing the CAR. In some embodiments, the immune-potentiation is inflammatory, e.g. signaling gives rise to increase in the potency of the immune cells bearing the CAR. In some embodiments, the immune-potentiation prevents exhaustion, e.g. signaling maintains the long-term functionality of immune cells bearing the CAR. 
     In some embodiments, the recruiting domains of the disclosure are synthetic, and do not resemble any naturally occurring receptor fragment. 
     In some embodiments, the Stat-recruiting domains of the disclosure are synthetic, and do not resemble any naturally occurring receptor fragment. 
     In some embodiments the recruiting domain is connected to the C-terminus of the JAK binding domain, either directly, or via a linker. In some embodiments, the linker can comprise one or more amino acid residues. 
     In other embodiments, the Stat-recruiting domains of the disclosure are derived from cytoplasmic tails of naturally occurring receptors, e.g. derived from naturally occurring cytokine receptors. These cytoplasmic tails of naturally occurring receptors may be the regions downstream of the JAK-activating domains of the transmembrane domain of the receptor. The Stat-recruiting domains of the chimeric cytokine receptors comprise at least one STAT-recruiting domain from at least one receptor. In some embodiments, the Stat-recruiting domain comprises at least one STAT1-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT2-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT3-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT4-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT5-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT6-recruiting domain. In some embodiments, the Stat-recruiting domain comprises at least one STAT7-recruiting domain. 
     In some embodiments, the naturally occurring receptor from which the Stat-recruiting domain is derived, is a not a cytokine receptor. 
     In some embodiments, the naturally occurring receptor from which the Stat-recruiting domain is derived, is a cytokine receptor. Exemplary cytokine receptors through which T-cell-immune potentiating cytokines signal include, but are not limited to IL-2 receptor, IL-7 receptor, IL-15 receptor and IL-21 receptor. In alternative embodiments, the receptor from which the Stat-recruiting domain is derived, is not a cytokine receptor. By choosing the Stat-recruiting domain of the chimeric cytokine receptor, the receptor can be redirected to signaling of choice. 
     Table 5 provides exemplary receptors from which Stat-recruiting domains of the chimeric cytokine receptors of the disclosure are derived. Table 6a provides exemplary amino acid sequences of recruiting domains of the disclosure. 
     In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 45. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 46. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 47. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 48. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 49. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 50. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 51. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 52. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 53. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 54. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 55. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 56. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 57. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 58. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 59. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 60. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 61. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 62. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 63. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 64. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 65. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 66. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 67. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 68. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 69. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 70. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 71. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 72. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 73. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 74. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 75. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 76. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 77. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 78. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 79. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 80. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 81. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 82. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 83. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 84. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 85. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 86. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 87. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 170. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of the STAT-recruiting domain of SEQ ID NO: 171. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises an amino acid sequence of the STAT-recruiting domain that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the amino acid sequence of any one of the sequences shown in Table 6a. 
     
       
         
           
               
             
               
                 TABLE 5 
               
               
                   
               
               
                 Source for recruiting domains 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                   
                 BLNK 
               
               
                   
                 IL2RG 
               
               
                   
                 EGFR 
               
               
                   
                 EpoR 
               
               
                   
                 GHR 
               
               
                   
                 IFNAR1 
               
               
                   
                 IFNAR2 
               
               
                   
                 IFNAR1/2 
               
               
                   
                 IFNLR1 
               
               
                   
                 IL10R1 
               
               
                   
                 IL12Rb1 
               
               
                   
                 IL12Rb2 
               
               
                   
                 IL21R 
               
               
                   
                 IL2Rb 
               
               
                   
                 IL2small 
               
               
                   
                 IL7R 
               
               
                   
                 IL7Ra 
               
               
                   
                 IL9R 
               
               
                   
                 IL15R 
               
               
                   
                 IL21R 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6a 
               
             
            
               
                   
               
               
                 Recruiting domain Sequences (Cytotail Sequences) 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ 
               
               
                 Recruiting domain 
                 Amino acid sequence 
                 ID NO: 
               
               
                   
               
               
                 IL7R(316-459) 
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPN 
                  45 
               
               
                   
                 CPSEDVVITPESFGRDSSLTCLAGNVSACDAPILS 
                   
               
               
                   
                 SSRSLDCRESGKNGPHVYQDLLLSLGTTNSTLPPP 
                   
               
               
                   
                 FSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMS 
                   
               
               
                   
                 SFYQNQ 
                   
               
               
                   
               
               
                 IL2Rb(333-551) 
                 VTQLLLQQDKVPEPASLSSNHSLTSCFTNQGYFF 
                  46 
               
               
                   
                 FHLPDALEIEACQVYFTYDPYSEEDPDEGVAGAP 
                   
               
               
                   
                 TGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSLL 
                   
               
               
                   
                 GGPSPPSTAPGGSGAGEERMPPSLQERVPRDWDP 
                   
               
               
                   
                 QPLGPPTPGVPDLVDFQPPPELVLREAGEEVPDA 
                   
               
               
                   
                 GPREGVSFPWSRPPGQGEFRALNARLPLNTDAYL 
                   
               
               
                   
                 SLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 IFNAR1(508-557) 
                 ISTIATVEETNQTDEDHKKYSSQTSQDSGNYSNE 
                  47 
               
               
                   
                 DESESKTSEELQQDFV 
                   
               
               
                   
               
               
                 IFNAR2(310-515) 
                 KKKVWDYNYDDESDSDTEAAPRTSGGGYTMHG 
                  48 
               
               
                   
                 LTVRPLGQASATSTESQLIDPESEEEPDLPEVDVE 
                   
               
               
                   
                 LPTMPKDSPQQLELLSGPCERRKSPLQDPFPEEDY 
                   
               
               
                   
                 SSTEGSGGRITFNVDLNSVFLRVLDDEDSDDLEA 
                   
               
               
                   
                 PLMLSSHLEEMVDPEDPDNVQSNHLLASGEGTQ 
                   
               
               
                   
                 PTFPSPSSEGLWSEDAPSDQSDTSESDVDLGDGYI 
                   
               
               
                   
                 MR 
                   
               
               
                   
               
               
                 IFNAR1/2(IFNAR1 
                 ISTIATVEETNQTDEDHKKYSSQTSQDSGNYSNE 
                  49 
               
               
                 residues 508-557-IFNAR2 
                 DESESKTSEELQQDFVKKKVWDYNYDDESDSDT 
                   
               
               
                 residues 310-515) 
                 EAAPRTSGGGYTMHGLTVRPLGQASATSTESQLI 
                   
               
               
                   
                 DPESEEEPDLPEVDVELPTMPKDSPQQLELLSGPC 
                   
               
               
                   
                 ERRKSPLQDPFPEEDYSSTEGSGGRITFNVDLNSV 
                   
               
               
                   
                 FLRVLDDEDSDDLEAPLMLSSHLEEMVDPEDPD 
                   
               
               
                   
                 NVQSNHLLASGEGTQPTFPSPSSEGLWSEDAPSD 
                   
               
               
                   
                 QSDTSESDVDLGDGYIMR 
                   
               
               
                   
               
               
                 IFNLR1(300-520) 
                 RGVRPTPRVRAPATQQTRWKKDLAEDEEEEDEE 
                  50 
               
               
                   
                 DTEDGVSFQPYIEPPSFLGQEHQAPGHSEAGGVD 
                   
               
               
                   
                 SGRPRAPLVPSEGSSAWDSSDRSWASTVDSSWD 
                   
               
               
                   
                 RAGSSGYLAEKGPGQGPGGDGHQESLPPPEFSKD 
                   
               
               
                   
                 SGFLEELPEDNLSSWATWGTLPPEPNLVPGGPPV 
                   
               
               
                   
                 SLQTLTFCWESSPEEEEEARESEIEDSDAGSWGAE 
                   
               
               
                   
                 STQRTEDRGRTLGHYMAR 
                   
               
               
                   
               
               
                 Common Gamma 
                 IPPKGGALGEGPGASPCNQHSPYWAPPCYTLKPE 
                  51 
               
               
                 Chain(335-369) 
                 T 
                   
               
               
                   
               
               
                 IL9R(356-521) 
                 TALLTCGPARPWKSVALEEEQEGPGTRLPGNLSS 
                  52 
               
               
                   
                 EDVLPAGCTEWRVQTLAYLPQEDWAPTSLTRPA 
                   
               
               
                   
                 PPDSEGSRSSSSSSSSNNNNYCALGCYGGWHLSA 
                   
               
               
                   
                 LPGNTQSSGPIPALACGLSCDHQGLETQQGVAW 
                   
               
               
                   
                 VLAGHCQRPGLHEDLQGMLLPSVLSKARSWTF 
                   
               
               
                   
               
               
                 IL21R(322-538) 
                 PRSPAKRLQLTELQEPAELVESDGVPKPSFWPTA 
                  53 
               
               
                   
                 QNSGGSAYSEERDRPYGLVSIDTVTVLDAEGPCT 
                   
               
               
                   
                 WPCSCEDDGYPALDLDAGLEPSPGLEDPLLDAG 
                   
               
               
                   
                 TTVLSCGCVSAGSPGLGGPLGSLLDRLKPPLADG 
                   
               
               
                   
                 EDWAGGLPWGGRSPGGVSESEAGSPLAGLDMD 
                   
               
               
                   
                 TFDSGFVGSDCSSPVECDFTSPGDEGPPRSYLRQ 
                   
               
               
                   
                 WVVIPPPLSSPGPQAS 
                   
               
               
                   
               
               
                 GHR(353-638) 
                 PDEKTEESDTDRLLSSDHEKSHSNLGVKDGDSGR 
                  54 
               
               
                   
                 TSCCEPDILETDFNANDIHEGTSEVAQPQRLKGE 
                   
               
               
                   
                 ADLLCLDQKNQNNSPYHDACPATQQPSVIQAEK 
                   
               
               
                   
                 NKPQPLPTEGAESTHQAAHIQLSNPSSLSNIDFYA 
                   
               
               
                   
                 QVSDITPAGSVVLSPGQKNKAGMSQCDMHPEM 
                   
               
               
                   
                 VSLCQENFLMDNAYFCEADAKKCIPVAPHIKVES 
                   
               
               
                   
                 HIQPSLNQEDIYITTESLTTAAGRPGTGEHVPGSE 
                   
               
               
                   
                 MPVPDYTSIHIVQSPQGLILNATALPLPDKEFLSS 
                   
               
               
                   
                 CGYVSTDQLNKIMP 
                   
               
               
                   
               
               
                 EpoR(339-508) 
                 WGTMQAVEPGTDDEGPLLEPVGSEHAQDTYLVL 
                  55 
               
               
                   
                 DKWLLPRNPPSEDLPGPGGSVDIVAMDEGSEASS 
                   
               
               
                   
                 CSSALASKPSPEGASAASFEYTILDPSSQLLRPWT 
                   
               
               
                   
                 LCPELPPTPPHLKYLYLVVSDSGISTDYSSGDSQG 
                   
               
               
                   
                 AQGGLSDGPYSNPYENSLIPAAEPLPPSYVACS 
                   
               
               
                   
               
               
                 murine IL2Rb(337-539) 
                 AVQLLLLQKDSAPLPSPSGHSQASCFTNQGYFFF 
                  56 
               
               
                   
                 HLPNALEIESCQVYFTYDPCVEEEVEEDGSRLPE 
                   
               
               
                   
                 GSPHPPLLPLAGEQDDYCAFPPRDDLLLFSPSLST 
                   
               
               
                   
                 PNTAYGGSRAPEERSPLSLHEGLPSLASRDLMGL 
                   
               
               
                   
                 QRPLERMPEGDGEGLSANSSGEQASVPEGNLHG 
                   
               
               
                   
                 QDQDRGQGPILTLNTDAYLSLQELQAQDSVHLI 
                   
               
               
                   
               
               
                 murine IL7Ra(316-459) 
                 RDEVESFLPNDLPAQPEELETQGHRAAVHSAN 
                  57 
               
               
                   
                 RSPETSVSPPETVRRESPLRCLARNLSTCNAPPLL 
                   
               
               
                   
                 SSRSPDYRDGDRNRPPVYQDLLPNSGNTNVPVPV 
                   
               
               
                   
                 PQPLPFQSGILIPVSQRQPISTSSVLNQEEAYVTMS 
                   
               
               
                   
                 SFYQNK 
                   
               
               
                   
               
               
                 EGFR(955-1186) 
                 VIQGDERMHLPSPTDSNFYRALMDEEDMDDVVD 
                  58 
               
               
                   
                 ADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA 
                   
               
               
                   
                 CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSID 
                   
               
               
                   
                 DTFLPVPEYINQSVPKRPAGSVQNPVYHNQPLNP 
                   
               
               
                   
                 APSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTF 
                   
               
               
                   
                 DSPAHWAQKGSHQISLDNPDYQQDFFPKEAKPN 
                   
               
               
                   
                 GIFKGSTAENAEYLRVAPQSSEFIGA 
                   
               
               
                   
               
               
                 EGFR(955-1186; 
                 VIQGDERMHLPSPTDSNFFRALMDEEDMDDVVD 
                  59 
               
               
                 Y974F, d1045-1057) 
                 ADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA 
                   
               
               
                   
                 CIDRNGLQSCPIKEDSFLQRIDDTFLPVPEYINQSV 
                   
               
               
                   
                 PKRPAGSVQNPVYHNQPLNPAPSRDPHYQDPHS 
                   
               
               
                   
                 TAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSH 
                   
               
               
                   
                 QISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEY 
                   
               
               
                   
                 LRVAPQSSEFIGA 
                   
               
               
                   
               
               
                 EGFR(955-1009; Y974F) 
                 VIQGDERMHLPSPTDSNFFRALMDEEDMDDVVD 
                  60 
               
               
                   
                 ADEYLIPQQGFFSSPSTSRTP 
                   
               
               
                   
               
               
                 EGFR(1019-1085) 
                 NNSTVACIDRNGLQSCPIKEDSFLQRIDDTFLPVP 
                  61 
               
               
                   
                 EYINQSVPKRPAGSVQNPV 
                   
               
               
                   
               
               
                 EGFR(1037-1103, 
                 KEDSFLQRIDDTFLPVPEFINQSVPKRPAGSVQNP 
                  62 
               
               
                 Y1068/1101F, 
                 VYHNQPLNPAPSRDPHFQD 
                   
               
               
                 d1045-1057) 
                   
                   
               
               
                   
               
               
                 EGFR(1066- 
                 VPEFINQSVPKRPAGSVQNPVFHNQPLNPAPSRD 
                  63 
               
               
                 1118, Y1068/1086F) 
                 PHYQDPHSTAVGNPEYLNTV 
                   
               
               
                   
               
               
                 EGFR(1122-1165) 
                 PEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDN 
                  64 
               
               
                   
                 PDYQQDFFPKEAKPNGIFKG 
                   
               
               
                   
               
               
                 EGFR(1133- 
                 WAQKGSHQISLDNPDFQQDFFPKEAKPNGIFKGS 
                  65 
               
               
                 1186, Y1148F) 
                 TAENAEYLRVAPQSSEFIGA 
                   
               
               
                   
               
               
                 IL12Rb2(775-825) 
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDL 
                  66 
               
               
                   
                 PSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 IL7R(376-416) 
                 ACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLG 
                  67 
               
               
                   
                 TTNSTLP 
                   
               
               
                   
               
               
                 IL7R(424-459) 
                 GILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQ 
                  68 
               
               
                   
                 NQ 
                   
               
               
                   
               
               
                 IL7R(376-416, 424-459) 
                 ACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLG 
                  69 
               
               
                   
                 TTNSTLPQGQPILTSLGSNQEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 IL7R(424-459, Y456F) 
                 GILTLNPVAQGQPILTSLGSNQEEAYVTMSSFFQN 
                  70 
               
               
                   
                 Q 
                   
               
               
                   
               
               
                 IL7R(376-416, 424- 
                 ACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLG 
                  71 
               
               
                 459, Y456F) 
                 TTNSTLPQGQPILTSLGSNQEEAYVTMSSFFQNQ 
                   
               
               
                   
               
               
                 IL2Rbsmall(393-433) 
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDD 
                  72 
               
               
                   
                 LLLFSPSGQGEFRALNARLPLNTDAYLSLQELQG 
                   
               
               
                   
                 QDPTHLV 
                   
               
               
                   
               
               
                 IL2Rbsmall(518-551) 
                 GQGEFRALNARLPLNTDAYLSLQELQGQDPTHL 
                  73 
               
               
                   
                 V 
                   
               
               
                   
               
               
                 IL2Rbsmall(339-379, 
                 QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDA 
                  74 
               
               
                 393-433) 
                 LEIEACQDEGVAGAPTGSSPQPLQPLSGEDDAYC 
                   
               
               
                   
                 TFPSRDDLLLFSPS 
                   
               
               
                   
               
               
                 IL2Rbsmall(339-379, 
                 QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDA 
                  75 
               
               
                 518-551) 
                 LEIEACQ 
                   
               
               
                   
                 GQGEFRALNARLPLNTDAYLSLQELQGQDPTHL 
                   
               
               
                   
                 V 
                   
               
               
                   
               
               
                 IL2Rbsmall(393-433, 
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDD 
                  76 
               
               
                 518-551) 
                 LLLFSPSGQGEFRALNARLPLNTDAYLSLQELQG 
                   
               
               
                   
                 QDPTHLV 
                   
               
               
                   
               
               
                 IL2Rbsmall(339-379, 
                 QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDA 
                  77 
               
               
                 393-433, 518-551) 
                 LEIEACQDEGVAGAPTGSSPQPLQPLSGEDDAYC 
                   
               
               
                   
                 TFPSRDDLLLFSPSGQGEFRALNARLPLNTDAYLS 
                   
               
               
                   
                 LQELQGQDPTHLV 
                   
               
               
                   
               
               
                 IFNAR2small(310-352) 
                 KKKVWDYNYDDESDSDTEAAPRTSGGGYTMHG 
                  78 
               
               
                   
                 LTVRPLGQASA 
                   
               
               
                   
               
               
                 IFNAR2small(486-515) 
                 EGLWSEDAPSDQSDTSESDVDLGDGYIMR 
                  79 
               
               
                   
               
               
                 IFNAR2small(310-352, 
                 KKKVWDYNYDDESDSDTEAAPRTSGGGYTMHG 
                  80 
               
               
                 486-515) 
                 LTVRPLGQASA 
                   
               
               
                   
                 EGLWSEDAPSDQSDTSESDVDLGDGYIMR 
                   
               
               
                   
               
               
                 BLNK(53-208) 
                 ASESPADEEEQWSDDFDSDYENPDEHSDSEMYV 
                  81 
               
               
                   
                 MPAEENADDSYEPPPVEQETRPVHPALPFARGEY 
                   
               
               
                   
                 IDNRSSQRHSPPFSKTLPSKPSWPSEKARLTSTLP 
                   
               
               
                   
                 ALTALQKPQVPPKPKGLLEDEADYVVPVEDNDE 
                   
               
               
                   
                 NYIHPTESSSPPPEKAPMVNR 
                   
               
               
                   
               
               
                 BLNK(53-208, Y72F) 
                 ASESPADEEEQWSDDFDSDFENPDEHSDSEMYV 
                  82 
               
               
                   
                 MPAEENADDSYEPPPVEQETRPVHPALPFARGEY 
                   
               
               
                   
                 IDNRSSQRHSPPFSKTLPSKPSWPSEKARLTSTLP 
                   
               
               
                   
                 ALTALQKPQVPPKPKGLLEDEADYVVPVEDNDE 
                   
               
               
                   
                 NYIHPTESSSPPPEKAPMVNR 
                   
               
               
                   
               
               
                 BLNK(53-208, 
                 ASESPADEEEQWSDDFDSDFENPDEHSDSEMYV 
                  83 
               
               
                 Y72F, Y96F) 
                 MPAEENADDSFEPPPVEQETRPVHPALPFARGEY 
                   
               
               
                   
                 IDNRSSQRHSPPFSKTLPSKPSWPSEKARLTSTLP 
                   
               
               
                   
                 ALTALQKPQVPPKPKGLLEDEADYVVPVEDNDE 
                   
               
               
                   
                 NYIHPTESSSPPPEKAPMVNR 
                   
               
               
                   
               
               
                 EpoR(339-508) 
                 WGTMQAVEPGTDDEGPLLEPVGSEHAQDTYLVL 
                  84 
               
               
                   
                 DKWLLPRNPPSEDLPGPGGSVDIVAMDEGSEASS 
                   
               
               
                   
                 CSSALASKPSPEGASAASFEYTILDPSSQLLRPWT 
                   
               
               
                   
                 LCPELPPTPPHLKYLYLVVSDSGISTDYSSGDSQG 
                   
               
               
                   
                 AQGGLSDGPYSNPYENSLIPAAEPLPPSYVACS 
                   
               
               
                   
               
               
                 IL12Rb2(714-862) 
                 VTPVFRHPPCSNWPQREKGIQGHQASEKDMMHS 
                  85 
               
               
                   
                 ASSPPPPRALQAESRQLVDLYKVLESRGSDPKPE 
                   
               
               
                   
                 NPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAP 
                   
               
               
                   
                 LADSLEELEPQHISLSVFPSSSLHPLTFSCGDKLTL 
                   
               
               
                   
                 DQLKMRCDSLML 
                   
               
               
                   
               
               
                 IL12Rb1(622-662) 
                 WDKGERTEPLEKTELPEGAPELALDTELSLEDGD 
                  86 
               
               
                   
                 RCKAKM 
                   
               
               
                   
               
               
                 IL10R1(304-578) 
                 VSPELKNLDLHGSTDSGFGSTKPSLQTEEPQFLLP 
                  87 
               
               
                   
                 DPHPQADRTLGNREPPVLGDSCSSGSSNSTDSGIC 
                   
               
               
                   
                 LQEPSLSPSTGPTWEQQVGSNSRGQDDSGIDLVQ 
                   
               
               
                   
                 NSEGRAGDTQGGSALGHHSPPEPEVPGEEDPAA 
                   
               
               
                   
                 VAFQGYLRQTRCAEEKATKTGCLEEESPLTDGL 
                   
               
               
                   
                 GPKFGRCLVDEAGLHPPALAKGYLKQDPLEMTL 
                   
               
               
                   
                 ASSGAPTGQWNQPTEEWSLLALSSCSDLGISDWS 
                   
               
               
                   
                 FAHDLAPLGCVAAPGGLLGSFNSDLVTLPLISSLQ 
                   
               
               
                   
                 SSE 
                   
               
               
                   
               
               
                 IL2Rb(333-551, 
                 VTQLLLQQDKVPEPASLSSNHSLTSCFTNQGYFF 
                 170 
               
               
                 Y381S, Y384S, Y387S) 
                 FHLPDALEIEACQV   S   FT   S   DP   S   SEEDPDEGVAGAPT 
                   
               
               
                   
                 GSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSLLG 
                   
               
               
                   
                 GPSPPSTAPGGSGAGEERMPPSLQERVPRDWDPQ 
                   
               
               
                   
                 PLGPPTPGVPDLVDFQPPPELVLREAGEEVPDAG 
                   
               
               
                   
                 PREGVSFPWSRPPGQGEFRALNARLPLNTDAYLS 
                   
               
               
                   
                 LQELQGQDPTHLV 
                   
               
               
                   
               
               
                 IL2Rb(333-551, 
                 VTQLLLQQDKVPEPASLSSNHSLTSCFTNQG   S   FFF 
                 171 
               
               
                 Y364S, Y381S, Y384S, 
                 HLPDALEIEACQV   S   FT   S   DP   S   SEEDPDEGVAGAPTG 
                   
               
               
                 Y387S) 
                 SSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSLLGG 
                   
               
               
                   
                 PSPPSTAPGGSGAGEERMPPSLQERVPRDWDPQP 
                   
               
               
                   
                 LGPPTPGVPDLVDFQPPPELVLREAGEEVPDAGP 
                   
               
               
                   
                 REGVSFPWSRPPGQGEFRALNARLPLNTDAYLSL 
                   
               
               
                   
                 QELQGQDPTHLV 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the Stat-recruiting domain of a chimeric cytokine receptor of the disclosure comprises a STAT-recruiting domain from one receptor. 
     In order to generate multiple outputs, one or more STAT-recruiting domains may be joined in tandem to mimic signaling from one or more cytokines. 
     In some embodiments, the STAT-recruiting domain comprises portions of more than one receptor, e.g. comprising more than one STAT-recruiting domain. In such embodiments, a tandem cytokine signaling domain is provided, allowing for enhanced signaling. Accordingly, in some embodiments, the STAT-recruiting domain of a monomer of the chimeric cytokine receptor of the disclosure comprises the STAT-recruiting domains from more than one receptor, e.g. comprises the STAT-recruiting domains from two, three, four, five, or even six receptors. For example, in some embodiments, STAT-recruiting domains can be linked in tandem to stimulate multiple pathways (e.g., the IL7R(316-459)-IL12Rb2(775-825) fragment fusion for pro-persistence STAT5 and pro-inflammatory STAT4; IL7R(316-459)-IL2Rbsmall(393-433, 518-551) for pro-persistence; IL7R(316-459)-EGFR(1122-1165) for pro-persistence and anti-exhaustion; IL2Rbsmall(393-433, 518-551)-EGFR(1122-1165) for pro-persistence and anti-exhaustion). 
     In some embodiments, the more than one STAT-recruiting domains can be connected either directly, or via a linker. In some embodiments, the linker can comprise one or more amino acid residues. 
     When generating multiple outputs, the proximity of individual STAT-recruiting domains to the cell membrane can influence the strength of their respective signaling outputs. Table 6b shows examples of PD-1 chimeric cytokine receptors with the dual outputs, where each output can be placed either proximal or distal to the cell membrane. 
     
       
         
           
               
             
               
                 TABLE 6b 
               
             
            
               
                   
               
               
                 Examples of PD-1 chimeric cytokine receptors with dual outputs 
               
            
           
           
               
               
               
            
               
                 Dual output STAT-recruiting 
                 Membrane  
                   
               
               
                 domain 
                 proximal 
                 Membrane distal 
               
               
                   
               
               
                 IL2Rbsmall(393-433, 518-551)/ 
                 IL2Rbsmall(393- 
                 IL21R(322-538) 
               
               
                 IL21R(322-538) 
                 433, 518-551) 
                   
               
               
                 IL21R(322-538)/ 
                 IL21R(322-538) 
                 IL2Rbsmall(393- 
               
               
                 IL2Rbsmall(393-433, 518-551) 
                   
                 433, 518-551) 
               
               
                 IL2Rbsmall(339-379, 393- 
                 IL2Rbsmall(339- 
                 IL21R(322-538) 
               
               
                 433, 518-551)/IL21R(322-538) 
                 379, 393-433, 
                   
               
               
                   
                 518-551) 
                   
               
               
                 IL21R(322-538)/ 
                 IL21R(322-538) 
                 IL2Rbsmall(339- 
               
               
                 IL2Rbsmall(339-379, 393- 
                   
                 379, 393- 
               
               
                 433, 518-551) 
                   
                 433, 518-551) 
               
               
                 IL7R (316-459)/IL21R(322-538) 
                 IL7R (316-459) 
                 IL21R(322-538) 
               
               
                 IL21R(322-538)/IL7R (316-459) 
                 IL21R(322-538) 
                 IL7R (316-459) 
               
               
                   
               
            
           
         
       
     
     Without being bound to theory or mechanism, in some embodiments, a JAK-protein (JAK1, JAK2, JAK3, or TYK2) is bound to a PD-1 chimeric cytokine receptor of the disclosure (comprising a PD-1 ectodomain, a transmembrane domain, a JAK-binding domain, and a recruiting domain). In some embodiments, in the presence of (e.g. binding to) PD-L1, PD-L2, or an anti-PD-1 antibody, the PD-1 chimeric cytokine receptor clusters and allows for the two bound JAK-proteins to become activated, which in turn phosphorylate tyrosine residues on the recruiting domain of the chimeric receptor. The phosphorylated recruiting domains are then capable of binding the recruited proteins (e.g. a phosphorylated STAT-recruiting domain binds a STAT-protein), which in turn effectuate transcription events in the nucleus. 
     E. Exemplary PD-1 Chimeric Cytokine Receptors 
     Table 7 shows exemplary context-dependent cytokine receptor sequences of the disclosure. The receptors may be expressed with a signal sequence, e.g. a CD8SS of SEQ ID NO: 1. 
     In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 88. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 89. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 90. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 91. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 92. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 93. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 94. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 95. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 96. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 97. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 98. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 99. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 100. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 101. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 102. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 103. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 104. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 105. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 106. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 107. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 108. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 109. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 110. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 111. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 112. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 113. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 114. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 115. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 116. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 117. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 118. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 119. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 120. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of SEQ ID NO: 121. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises the amino acid sequence of any one of SEQ ID NOs: 88-121, SEQ ID NOs: 136-145, and SEQ ID NOs: 172-213. In some embodiments, the PD-1 chimeric cytokine receptor of the disclosure comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOs: 88-121, SEQ ID NOs: 172-213. 
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Exemplary PD1 chimeric cytokine receptor sequences (assembled PD1 chimeric 
               
               
                 cytokine receptors): 
               
            
           
           
               
               
               
            
               
                 PD1 
                   
                   
               
               
                 chimeric 
                   
                   
               
               
                 cytokine 
                   
                 SEQ 
               
               
                 receptor 
                 Amino acid sequence 
                 ID NO: 
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  88 
               
               
                 207).TpoR 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (478- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 582).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                 (316- 
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                 459).IL12Rb2 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                 (775-825) 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  89 
               
               
                 207).GCSFR 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (614- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 710).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                 (316- 
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                 459).IL12Rb2 
                 LTLMTLTPEGSELHIILGLFGLLLLLTCLCGTAWLCCSPNRKN 
                   
               
               
                 (775-825) 
                 PLWPSVPDPAHSSLGSWVPTIMEEDAFQLPGLGTPPITKLTVL 
                   
               
               
                   
                 EEDEKKPVPWE 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  90 
               
               
                 207).GP130 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (609- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 700).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                 (316- 
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                 459).IL12Rb2 
                 TTPKFAQGEIEAIVVPVCLAFLLTTLLGVLFCFNKRDLIKKHI 
                   
               
               
                 (775-825) 
                 WPNVPDPSKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVS 
                   
               
               
                   
                 VVEIEAND 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 2xWTPD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  91 
               
               
                 (21-207) 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 .TpoR 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 (478- 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                 582).IL7Ra 
                 LAVICSRAARGTIGARRTGQPGWFLDSPDRPWNPPTFSPALL 
                   
               
               
                 (316- 
                 VVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPE 
                   
               
               
                 459).IL12Rb2 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCG 
                   
               
               
                 (775-825) 
                 AISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQ 
                   
               
               
                   
                 TLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 3xWTPD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  92 
               
               
                 (21- 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 207).TpoR 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 (478- 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                 582).IL7Ra 
                 LAVICSRAARGTIGARRTGQPGWFLDSPDRPWNPPTFSPALL 
                   
               
               
                 (316- 
                 VVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPE 
                   
               
               
                 459).IL12Rb2 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCG 
                   
               
               
                 (775-825) 
                 AISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQ 
                   
               
               
                   
                 TLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQP 
                   
               
               
                   
                 GWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                   
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                   
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                   
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                   
                 LAVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  93 
               
               
                 (478- 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 582).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  94 
               
               
                 (N-1) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETWISLVTALHLVLGLSAVLGLLLLRWQFPAH 
                   
               
               
                   
                 YRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCE 
                   
               
               
                   
                 EVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  95 
               
               
                 (N-2) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETISLVTALHLVLGLSAVLGLLLLRWQFPAHY 
                   
               
               
                   
                 RRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEE 
                   
               
               
                   
                 VEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  96 
               
               
                 (N-2+1) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLISLVTALHLVLGLSAVLGLLLLRWQFPAH 
                   
               
               
                   
                 YRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCE 
                   
               
               
                   
                 EVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  97 
               
               
                 (N-3) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETSLVTALHLVLGLSAVLGLLLLRWQFPAHYR 
                   
               
               
                   
                 RLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEV 
                   
               
               
                   
                 EPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  98 
               
               
                 (N-4) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLVTALHLVLGLSAVLGLLLLRWQFPAHYR 
                   
               
               
                   
                 RLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEV 
                   
               
               
                   
                 EPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                  99 
               
               
                 (N-4+1) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETILVTALHLVLGLSAVLGLLLLRWQFPAHYR 
                   
               
               
                   
                 RLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEV 
                   
               
               
                   
                 EPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 100 
               
               
                 (N-5) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETVTALHLVLGLSAVLGLLLLRWQFPAHYRR 
                   
               
               
                   
                 LRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEP 
                   
               
               
                   
                 SLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 101 
               
               
                 (N-6) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETTALHLVLGLSAVLGLLLLRWQFPAHYRRLR 
                   
               
               
                   
                 HALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSL 
                   
               
               
                   
                 LEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 102 
               
               
                 (N-7) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETALHLVLGLSAVLGLLLLRWQFPAHYRRLR 
                   
               
               
                   
                 HALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSL 
                   
               
               
                   
                 LEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 103 
               
               
                 (N-8) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLHLVLGLSAVLGLLLLRWQFPAHYRRLRH 
                   
               
               
                   
                 ALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLL 
                   
               
               
                   
                 EILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 104 
               
               
                 (N-9) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETHLVLGLSAVLGLLLLRWQFPAHYRRLRHA 
                   
               
               
                   
                 LWPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEI 
                   
               
               
                   
                 LPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 105 
               
               
                 (N-10) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLVLGLSAVLGLLLLRWQFPAHYRRLRHAL 
                   
               
               
                   
                 WPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEIL 
                   
               
               
                   
                 PKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 106 
               
               
                 (N-11) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETVLGLSAVLGLLLLRWQFPAHYRRLRHALW 
                   
               
               
                   
                 PSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILP 
                   
               
               
                   
                 KSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 107 
               
               
                 (N-12) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLGLSAVLGLLLLRWQFPAHYRRLRHALWP 
                   
               
               
                   
                 SLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPK 
                   
               
               
                   
                 SSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 108 
               
               
                 (N-13) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETGLSAVLGLLLLRWQFPAHYRRLRHALWPS 
                   
               
               
                   
                 LPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKS 
                   
               
               
                   
                 SERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 109 
               
               
                 (N-14) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLSAVLGLLLLRWQFPAHYRRLRHALWPSLP 
                   
               
               
                   
                 DLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 110 
               
               
                 (N-15) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETSAVLGLLLLRWQFPAHYRRLRHALWPSLP 
                   
               
               
                   
                 DLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 111 
               
               
                 (N-16) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETAVLGLLLLRWQFPAHYRRLRHALWPSLPD 
                   
               
               
                   
                 LHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSER 
                   
               
               
                   
                 TPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 112 
               
               
                 (N-17) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETVLGLLLLRWQFPAHYRRLRHALWPSLPDL 
                   
               
               
                   
                 HRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERT 
                   
               
               
                   
                 PLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 113 
               
               
                 (N-18) 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 .IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 459).IL12Rb2 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                 (775-825) 
                 SDPTRVETATETLGLLLLRWQFPAHYRRLRHALWPSLPDLH 
                   
               
               
                   
                 RVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTP 
                   
               
               
                   
                 LPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 114 
               
               
                 (N+1).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWLISLVTALHLVLGLSAVLGLLLLRWQFP 
                   
               
               
                   
                 AHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDT 
                   
               
               
                   
                 CEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 115 
               
               
                 (N+2).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWVLISLVTALHLVLGLSAVLGLLLLRWQF 
                   
               
               
                   
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                   
                 TCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 116 
               
               
                 (N+3).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWLVLISLVTALHLVLGLSAVLGLLLLRWQ 
                   
               
               
                   
                 FPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVS 
                   
               
               
                   
                 DTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 117 
               
               
                 (N+4).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWILVLISLVTALHLVLGLSAVLGLLLLRW 
                   
               
               
                   
                 QFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATV 
                   
               
               
                   
                 SDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 118 
               
               
                 (N+5).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWLILVLISLVTALHLVLGLSAVLGLLLLR 
                   
               
               
                   
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKAT 
                   
               
               
                   
                 VSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 119 
               
               
                 (N+6).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWLLILVLISLVTALHLVLGLSAVLGLLLLR 
                   
               
               
                   
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKAT 
                   
               
               
                   
                 VSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 120 
               
               
                 (N+7).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWVLLILVLISLVTALHLVLGLSAVLGLLLL 
                   
               
               
                   
                 RWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKA 
                   
               
               
                   
                 TVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 121 
               
               
                 (N+8).IL7Ra 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (316- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 459).IL12Rb2 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                 (775-825) 
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
                 SDPTRVETATETAWLVLLILVLISLVTALHLVLGLSAVLGLLL 
                   
               
               
                   
                 LRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPK 
                   
               
               
                   
                 ATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
                 SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLA 
                   
               
               
                   
                 DSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 136 
               
               
                 170).TPOR/ 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 MPLR(478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, H499L, 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 S505N, W515K) 
                 SDPTRVETATETAWISLVTAL   L   LVLGL   N   AVLGLLLLR   K   QFPA 
                   
               
               
                 IL7Ra 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                 (316-459) 
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 137 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, S505N, 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 W515K), 
                 SDPTRVETATETAWISLVTALHLVLGL   N   AVLGLLLLR   K   QFPA 
                   
               
               
                 IL2small(393- 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                 433, 518-551) 
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1DN 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 138 
               
               
                 (HAPD1 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 Dominant 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 Negative) 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                   
                 AVICSRAARGTIGARRTGQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 139 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 (316-459) 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 140 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582).IL2small 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 (393- 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                 433, 518-551) 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 172 
               
               
                 170).TpoR 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (478- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 582).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVT 
                   
               
               
                 (316- 
                 ALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHR 
                   
               
               
                 459).IL12Rb2 
                 VLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPL 
                   
               
               
                 (775-825) 
                 PL LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSED 
                   
               
               
                   
                 VVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKN 
                   
               
               
                   
                 GPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTS 
                   
               
               
                   
                 LGSNQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPA 
                   
               
               
                   
                 GDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 173 
               
               
                 170).GCSFR 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (614- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 710).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVLTLMTLTPEGSELHIILGLF 
                   
               
               
                 (316- 
                 GLLLLLTCLCGTAWLCCSPNRKNPLWPSVPDPAHSSLGSWV 
                   
               
               
                 459).IL12Rb2 
                 PTIMEEDAFQLPGLGTPPITKLTVLEEDEKKPVPWE LE ARDEV 
                   
               
               
                 (775-825) 
                 EGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGR 
                   
               
               
                   
                 DSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLL 
                   
               
               
                   
                 LSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAY 
                   
               
               
                   
                 VTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGY 
                   
               
               
                   
                 LPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 WTPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 174 
               
               
                 170).GP130 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 (609- 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 700).IL7Ra 
                 RRAEVPTAHPSPSPRPAGQFQTLVTTPKFAQGEIEAIVVPVCL 
                   
               
               
                 (316- 
                 AFLLTTLLGVLFCFNKRDLIKKHIWPNVPDPSKSHIAQWSPHT 
                   
               
               
                 459).IL12Rb2 
                 PPRHNFNSKDQMYSDGNFTDVSVVEIEAND LE ARDEVEGFL 
                   
               
               
                 (775-825) 
                 QDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSL 
                   
               
               
                   
                 TCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLG 
                   
               
               
                   
                 TTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTM 
                   
               
               
                   
                 SSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLPSN 
                   
               
               
                   
                 IDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 2xWTPD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 175 
               
               
                 (21- 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 170).TpoR 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 (478- 
                 RRAEVPTAHPSPSPRPAGQFQTLV G PGWFLDSPDRPWNPPTF 
                   
               
               
                 582).IL7Ra 
                 SPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKL 
                   
               
               
                 (316- 
                 AAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGT 
                   
               
               
                 459).IL12Rb2 
                 YLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPA 
                   
               
               
                 (775-825) 
                 GQFQTLVSDPTRVETATETAWISLVTALHLVLGLSAVLGLLL 
                   
               
               
                   
                 LRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPK 
                   
               
               
                   
                 ATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFLQDTF 
                   
               
               
                   
                 PQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLA 
                   
               
               
                   
                 GNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNS 
                   
               
               
                   
                 TLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFY 
                   
               
               
                   
                 QNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDL 
                   
               
               
                   
                 PSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 3xWTPD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 176 
               
               
                 (21- 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 170).TpoR 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 (478- 
                 RRAEVPTAHPSPSPRPAGQFQTLV G PGWFLDSPDRPWNPPTF 
                   
               
               
                 582).IL7Ra 
                 SPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKL 
                   
               
               
                 (316- 
                 AAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGT 
                   
               
               
                 459).IL12Rb2 
                 YLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPA 
                   
               
               
                 (775-825) 
                 GQFQTLV G PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTC 
                   
               
               
                   
                 SFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFR 
                   
               
               
                   
                 VTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESL 
                   
               
               
                   
                 RAELRVTERRAEVPTAHPSPSPRPAGQFQTLVSDPTRVETAT 
                   
               
               
                   
                 ETAWISLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHAL 
                   
               
               
                   
                 WPSLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEIL 
                   
               
               
                   
                 PKSSERTPLPL LE ARDEVEGFLQDTFPQQLEESEKQRLGGDV 
                   
               
               
                   
                 QSPNCPSEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSL 
                   
               
               
                   
                 DCRESGKNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPV 
                   
               
               
                   
                 AQGQPILTSLGSNQEEAYVTMSSFYQNQ GSGSR SDPKPENPA 
                   
               
               
                   
                 CPWTVLPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 177 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 (316- 
                 LHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 459).IL12Rb2 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 (775-825) 
                   LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVV 
                   
               
               
                   
                 ITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGP 
                   
               
               
                   
                 HVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLG 
                   
               
               
                   
                 SNQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGD 
                   
               
               
                   
                 LPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 178 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N-1) 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 .IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETWISLVTAL 
                   
               
               
                 (316- 
                 HLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVL 
                   
               
               
                 459).IL12Rb2 
                 GQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL L   
                   
               
               
                 (775-825) 
                   E ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVI 
                   
               
               
                   
                 TPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPH 
                   
               
               
                   
                 VYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGS 
                   
               
               
                   
                 NQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDL 
                   
               
               
                   
                 PTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 179 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 2).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETISLVTALH 
                   
               
               
                 (316- 
                 LVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLG 
                   
               
               
                 459).IL12Rb2 
                 QYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE   
                   
               
               
                 (775-825) 
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLP 
                   
               
               
                   
                 THDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 180 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 2+1).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLISLVTAL 
                   
               
               
                 (316- 
                 HLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVL 
                   
               
               
                 459).IL12Rb2 
                 GQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL L   
                   
               
               
                 (775-825) 
                   E ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVI 
                   
               
               
                   
                 TPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPH 
                   
               
               
                   
                 VYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGS 
                   
               
               
                   
                 NQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDL 
                   
               
               
                   
                 PTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 181 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 3).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETSLVTALHL 
                   
               
               
                 (316- 
                 VLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQ 
                   
               
               
                 459).IL12Rb2 
                 YLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE A 
                   
               
               
                 (775-825) 
                 RDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITP 
                   
               
               
                   
                 ESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVY 
                   
               
               
                   
                 QDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQ 
                   
               
               
                   
                 EEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPT 
                   
               
               
                   
                 HDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 182 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 4).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLVTALHL 
                   
               
               
                 (316- 
                 VLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQ 
                   
               
               
                 459).IL12Rb2 
                 YLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE A 
                   
               
               
                 (775-825) 
                 RDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITP 
                   
               
               
                   
                 ESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVY 
                   
               
               
                   
                 QDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQ 
                   
               
               
                   
                 EEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPT 
                   
               
               
                   
                 HDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 183 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 4+1).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETILVTALHL 
                   
               
               
                 (316- 
                 VLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQ 
                   
               
               
                 459).IL12Rb2 
                 YLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE A 
                   
               
               
                 (775-825) 
                 RDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITP 
                   
               
               
                   
                 ESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVY 
                   
               
               
                   
                 QDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQ 
                   
               
               
                   
                 EEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPT 
                   
               
               
                   
                 HDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 184 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 5).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETVTALHLV 
                   
               
               
                 (316- 
                 LGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQY 
                   
               
               
                 459).IL12Rb2 
                 LRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE AR 
                   
               
               
                 (775-825) 
                 DEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPES 
                   
               
               
                   
                 FGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQ 
                   
               
               
                   
                 DLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQE 
                   
               
               
                   
                 EAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTH 
                   
               
               
                   
                 DGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 185 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 6).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETTALHLVL 
                   
               
               
                 (316- 
                 GLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYL 
                   
               
               
                 459).IL12Rb2 
                 RDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARD 
                   
               
               
                 (775-825) 
                 EVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESF 
                   
               
               
                   
                 GRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQD 
                   
               
               
                   
                 LLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEE 
                   
               
               
                   
                 AYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHD 
                   
               
               
                   
                 GYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 186 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 7).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETALHLVLG 
                   
               
               
                 (316- 
                 LSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                 459).IL12Rb2 
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDE 
                   
               
               
                 (775-825) 
                 VEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFG 
                   
               
               
                   
                 RDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDL 
                   
               
               
                   
                 LLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEA 
                   
               
               
                   
                 YVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDG 
                   
               
               
                   
                 YLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 187 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 8).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLHLVLGLS 
                   
               
               
                 (316- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 459).IL12Rb2 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVE 
                   
               
               
                 (775-825) 
                 GFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRD 
                   
               
               
                   
                 SSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLS 
                   
               
               
                   
                 LGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVT 
                   
               
               
                   
                 MSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLP 
                   
               
               
                   
                 SNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 188 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 9).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETHLVLGLS 
                   
               
               
                 (316- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 459).IL12Rb2 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVE 
                   
               
               
                 (775-825) 
                 GFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRD 
                   
               
               
                   
                 SSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLS 
                   
               
               
                   
                 LGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVT 
                   
               
               
                   
                 MSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLP 
                   
               
               
                   
                 SNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 189 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 10).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLVLGLSA 
                   
               
               
                 (316- 
                 VLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTA 
                   
               
               
                 459).IL12Rb2 
                 ALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEG 
                   
               
               
                 (775-825) 
                 FLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDS 
                   
               
               
                   
                 SLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLS 
                   
               
               
                   
                 LGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVT 
                   
               
               
                   
                 MSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLP 
                   
               
               
                   
                 SNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 190 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 11).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETVLGLSAV 
                   
               
               
                 (316- 
                 LGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAA 
                   
               
               
                 459).IL12Rb2 
                 LSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGF 
                   
               
               
                 (775-825) 
                 LQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSS 
                   
               
               
                   
                 LTCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSL 
                   
               
               
                   
                 GTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVT 
                   
               
               
                   
                 MSSFYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLP 
                   
               
               
                   
                 SNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 191 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 12).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLGLSAVL 
                   
               
               
                 (316- 
                 GLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAAL 
                   
               
               
                 459).IL12Rb2 
                 SPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFL 
                   
               
               
                 (775-825) 
                 QDTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSL 
                   
               
               
                   
                 TCLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLG 
                   
               
               
                   
                 TTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTM 
                   
               
               
                   
                 SSFYQNQGSGSRSDPKPENPACPWTVLPAGDLPTHDGYLPSN 
                   
               
               
                   
                 IDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 192 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 13).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETGLSAVLG 
                   
               
               
                 (316- 
                 LLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALS 
                   
               
               
                 459).IL12Rb2 
                 PPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFLQ 
                   
               
               
                 (775-825) 
                 DTFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLT 
                   
               
               
                   
                 CLAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGT 
                   
               
               
                   
                 TNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSS 
                   
               
               
                   
                 FYQNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLPSNID 
                   
               
               
                   
                 DLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 193 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 14).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLSAVLGLL 
                   
               
               
                 (316- 
                 LLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPP 
                   
               
               
                 459).IL12Rb2 
                 KATVSDTCEEVEPSLLEILPKSSERTPLPLLEARDEVEGFLQD 
                   
               
               
                 (775-825) 
                 TFPQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTC 
                   
               
               
                   
                 LAGNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTT 
                   
               
               
                   
                 NSTLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSF 
                   
               
               
                   
                 YQNQGSGSRSDPKPENPACPWTVLPAGDLPTHDGYLPSNIDD 
                   
               
               
                   
                 LPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 194 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 15).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETSAVLGLLL 
                   
               
               
                 (316- 
                 LRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPK 
                   
               
               
                 459).IL12Rb2 
                 ATVSDTCEEVEPSLLEILPKSSERTPLPLLEARDEVEGFLQDTF 
                   
               
               
                 (775-825) 
                 PQQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLA 
                   
               
               
                   
                 GNVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNS 
                   
               
               
                   
                 TLPPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFY 
                   
               
               
                   
                 QNQ GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDL 
                   
               
               
                   
                 PSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 195 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 16).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAVLGLLLL 
                   
               
               
                 (316- 
                 RWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKA 
                   
               
               
                 459).IL12Rb2 
                 TVSDTCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFLQDTFP 
                   
               
               
                 (775-825) 
                 QQLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLAG 
                   
               
               
                   
                 NVSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNSTL 
                   
               
               
                   
                 PPPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQN 
                   
               
               
                   
                 QGSGSRSDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPS 
                   
               
               
                   
                 HEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 196 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 17).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETVLGLLLLR 
                   
               
               
                 (316- 
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKAT 
                   
               
               
                 459).IL12Rb2 
                 VSDTCEEVEPSLLEILPKSSERTPLPLLEARDEVEGFLQDTFPQ 
                   
               
               
                 (775-825) 
                 QLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLAGN 
                   
               
               
                   
                 VSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNSTLP 
                   
               
               
                   
                 PPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQNQ 
                   
               
               
                   
                 GSGSRSDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSH 
                   
               
               
                   
                 EAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 197 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 18).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLGLLLLR 
                   
               
               
                 (316- 
                 WQFPAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKAT 
                   
               
               
                 459).IL12Rb2 
                 VSDTCEEVEPSLLEILPKSSERTPLPLLEARDEVEGFLQDTFPQ 
                   
               
               
                 (775-825) 
                 QLEESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLAGN 
                   
               
               
                   
                 VSACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNSTLP 
                   
               
               
                   
                 PPFSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQNQ 
                   
               
               
                   
                   GSGSR SDPKPENPACPWTVLPAGDLPTHDGYLPSNIDDLPSH 
                   
               
               
                   
                 EAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 198 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+1).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWLISLVT 
                   
               
               
                 459).IL12Rb2 
                 ALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHR 
                   
               
               
                 (775-825) 
                 VLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPL 
                   
               
               
                   
                 PL LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSED 
                   
               
               
                   
                 VVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKN 
                   
               
               
                   
                 GPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTS 
                   
               
               
                   
                 LGSNQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLPA 
                   
               
               
                   
                 GDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 199 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+2).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWVLISLV 
                   
               
               
                 459).IL12Rb2 
                 TALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLH 
                   
               
               
                 (775-825) 
                 RVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTP 
                   
               
               
                   
                 LPLLEARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSE 
                   
               
               
                   
                 DVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGK 
                   
               
               
                   
                 NGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPIL 
                   
               
               
                   
                 TSLGSNQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPWTVLP 
                   
               
               
                   
                 AGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 200 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+3).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWLVLISL 
                   
               
               
                 459).IL12Rb2 
                 VTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDL 
                   
               
               
                 (775-825) 
                 HRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERT 
                   
               
               
                   
                 PLPLLEARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPS 
                   
               
               
                   
                 EDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESG 
                   
               
               
                   
                 KNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPI 
                   
               
               
                   
                 LTSLGSNQEEAYVTMSSFYQNQGSGSRSDPKPENPACPWTV 
                   
               
               
                   
                 LPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 201 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+4).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWILVLIS 
                   
               
               
                 459).IL12Rb2 
                 LVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPD 
                   
               
               
                 (775-825) 
                 LHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSER 
                   
               
               
                   
                 TPLPLLEARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCP 
                   
               
               
                   
                 SEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESG 
                   
               
               
                   
                 KNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPI 
                   
               
               
                   
                 LTSLGSNQEEAYVTMSSFYQNQGSGSRSDPKPENPACPWTV 
                   
               
               
                   
                 LPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 202 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+5).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWLILVLI 
                   
               
               
                 459).IL12Rb2 
                 SLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLP 
                   
               
               
                 (775-825) 
                 DLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPL LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNC 
                   
               
               
                   
                 PSEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRES 
                   
               
               
                   
                 GKNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQP 
                   
               
               
                   
                 ILTSLGSNQEEAYVTMSSFYQNQGSGSRSDPKPENPACPWTV 
                   
               
               
                   
                 LPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 203 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+6).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWLLILVL 
                   
               
               
                 459).IL12Rb2 
                 ISLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLP 
                   
               
               
                 (775-825) 
                 DLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSE 
                   
               
               
                   
                 RTPLPLLEARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNC 
                   
               
               
                   
                 PSEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRES 
                   
               
               
                   
                 GKNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQP 
                   
               
               
                   
                 ILTSLGSNQEEAYVTMSSFYQNQGSGSRSDPKPENPACPWTV 
                   
               
               
                   
                 LPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 204 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+7).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWVLLIL 
                   
               
               
                 459).IL12Rb2 
                 VLISLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWP 
                   
               
               
                 (775-825) 
                 SLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPK 
                   
               
               
                   
                 SSERTPLPLLEARDEVEGFLQDTFPQQLEESEKQRLGGDVQSP 
                   
               
               
                   
                 NCPSEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCR 
                   
               
               
                   
                 ESGKNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQG 
                   
               
               
                   
                 QPILTSLGSNQEEAYVTMSSFYQNQGSGSRSDPKPENPACPW 
                   
               
               
                   
                 TVLPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 205 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N+8).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWLVLLIL 
                   
               
               
                 459).IL12Rb2 
                 VLISLVTALHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWP 
                   
               
               
                 (775-825) 
                 SLPDLHRVLGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPK 
                   
               
               
                   
                 SSERTPLPL LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSP 
                   
               
               
                   
                 NCPSEDVVITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCR 
                   
               
               
                   
                 ESGKNGPHVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQG 
                   
               
               
                   
                 QPILTSLGSNQEEAYVTMSSFYQNQ GSGSR SDPKPENPACPW 
                   
               
               
                   
               
               
                   
                 TVLPAGDLPTHDGYLPSNIDDLPSHEAPLADSLEELEPQ 
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 206 
               
               
                 107).TPOR/ 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 MPLR(478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, H499L, 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 S505N, W515K) 
                 LLLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 .IL7Ra 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 (316-459) 
                   LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVV 
                   
               
               
                   
                 ITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGP 
                   
               
               
                   
                 HVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLG 
                   
               
               
                   
                 SNQEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 207 
               
               
                 (478- 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 582, S505N, 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 W515K) 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 .IL2Rbsmall 
                 LHLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 (393-433, 518- 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 551) 
                   LE DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPS 
                   
               
               
                   
                 GQGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 208 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582).IL7Ra 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 (316-459) 
                 LHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                   
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                   LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVV 
                   
               
               
                   
                 ITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGP 
                   
               
               
                   
                 HVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLG 
                   
               
               
                   
                 SNQEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 209 
               
               
                 (478- 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 582). 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 IL2Rbsmall(393- 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 433, 518-551) 
                 LHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                   
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                   LE DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPS 
                   
               
               
                   
                 GQGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 214 
               
               
                 107)TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, S505N, 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 W515K). 
                 LHLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 IL2Rbsmall(339- 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 379, 393- 
                   LE QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQ 
                   
               
               
                 433, 518-551) 
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 215 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, H499L, 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 S505N, W515K). 
                 LLLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 IL2Rbsmall 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 (339- 
                   LE QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQ 
                   
               
               
                 379,393- 
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                 433, 518-551) 
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 216 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478-582) 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 (339-379, 393- 
                 LHLVLGLSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 433, 518-551) 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                   LE QQDKVPEPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQ 
                   
               
               
                   
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 217 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N-7) 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 .IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETALHLVLG 
                   
               
               
                 (393- 
                 LSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                 433, 518-551) 
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE DEGV 
                   
               
               
                   
                 AGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFR 
                   
               
               
                   
                 ALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 218 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N-8) 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 .IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLHLVLGLS 
                   
               
               
                 (393- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 433, 518-551) 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE DEGVAG 
                   
               
               
                   
                 APTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFRAL 
                   
               
               
                   
                 NARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 219 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 9).IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETHLVLGLS 
                   
               
               
                 (393- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 433, 518-551) 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE DEGVAG 
                   
               
               
                   
                 APTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFRAL 
                   
               
               
                   
                 NARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 220 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 7).IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETALHLVLG 
                   
               
               
                 (339- 
                 LSAVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLR 
                   
               
               
                 379, 393- 
                 DTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE QQDK 
                   
               
               
                 433,518-551) 
                 VPEPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQDEGVAG 
                   
               
               
                   
                 APTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFRAL 
                   
               
               
                   
                 NARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 221 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 8).IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETLHLVLGLS 
                   
               
               
                 (339- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 379, 393- 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE QQDKVP 
                   
               
               
                 433, 518-551) 
                 EPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQDEGVAGAP 
                   
               
               
                   
                 TGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFRALNA 
                   
               
               
                   
                 RLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 222 
               
               
                 107).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (N- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 9).IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETHLVLGLS 
                   
               
               
                 (339- 
                 AVLGLLLLRWQFPAHYRRLRHALWPSLPDLHRVLGQYLRDT 
                   
               
               
                 379,393- 
                 AALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL LE QQDKVP 
                   
               
               
                 433, 518-551) 
                 EPASLSSNHSLTSCFTNQGYFFFHLPDALEIEACQDEGVAGAP 
                   
               
               
                   
                 TGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSGQGEFRALNA 
                   
               
               
                   
                 RLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 223 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478- 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 582, H499L, 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 S505N, W515K) 
                 LLLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 .IL2Rbsmall 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                 (393- 
                   LE DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPS 
                   
               
               
                 433, 518-551) 
                 GQGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 HAPD1(21- 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                 224 
               
               
                 170).TpoR 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 (478-582, 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 S505N, W515K) 
                 RAEVPTAHPSPSPRPAGQFQTLVSDPTRVETATETAWISLVTA 
                   
               
               
                 .IL7Ra 
                 LHLVLGLNAVLGLLLLRKQFPAHYRRLRHALWPSLPDLHRV 
                   
               
               
                 (316-459) 
                 LGQYLRDTAALSPPKATVSDTCEEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                   LE ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVV 
                   
               
               
                   
                 ITPESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGP 
                   
               
               
                   
                 HVYQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLG 
                   
               
               
                   
                 SNQEEAYVTMSSFYQNQ 
               
               
                   
               
            
           
         
       
     
     F. Expression of PD-1 Chimeric Cytokine Receptors 
     Provided herein are polynucleotides encoding any one of the chimeric cytokine receptors provided herein. Likewise, provided herein are expression vectors comprising such polynucleotides. In some embodiments, the vector is a viral vector. In some embodiments, the vector is not a viral vector. 
     In some embodiments, the vector comprises a polynucleotide encoding a PD-1 chimeric cytokine receptor, and a polynucleotide expressing a chimeric antigen receptor (CAR). 
     In some embodiments, expression of the chimeric cytokine receptor and the CAR are expressed as a single polypeptide chain, separated by a linker.  FIGS. 1B and 1D  show schematics of a vector that can be used to co-express the chimeric cytokine receptor and CAR of the disclosure. One or more STAT-recruiting domains may be joined in tandem to mimic signaling from one or more cytokines. 
     II. CAR-Bearing Immune Cells 
     Provided herein are engineered immune cells comprising a polynucleotide encoding a chimeric antigen receptor and a PD-1 chimeric cytokine receptor of the disclosure; and provided herein are engineered immune cells expressing a chimeric antigen receptor (CAR-I cell) and a PD-1 chimeric cytokine receptor of the disclosure. Examples of immune cells include T-cells, e.g., alpha/beta T-cells and gamma/delta T-cells, B cells, natural killer (NK) cells, natural killer T (NKT) cells, invariant NKT cells, mast cells, myeloic-derived phagocytes, dendritic cells, killer dendritic cells, macrophages, and monocytes. Immune cells also refer to cells derived from, for example without limitation, a stem cell. The stem cells can be adult stem cells, non-human embryonic stem cells, more particularly non-human stem cells, cord blood stem cells, progenitor cells, bone marrow stem cells, induced pluripotent stem cells, totipotent stem cells or hematopoietic stem cells. 
     Accordingly in some embodiments, provided herein are CAR-T-cells comprising a PD-1 chimeric cytokine receptor of the disclosure. 
     In some embodiments, a CAR can comprise an extracellular ligand-binding domain (e.g., a single chain variable fragment (scFv)), a transmembrane domain, and an intracellular signaling domain. In some embodiments, the extracellular ligand-binding domain, transmembrane domain, and intracellular signaling domain are in one polypeptide, i.e., in a single chain. Multichain CARs and polypeptides are also provided herein. In some embodiments, the multichain CARs comprise: a first polypeptide comprising a transmembrane domain and at least one extracellular ligand-binding domain, and a second polypeptide comprising a transmembrane domain and at least one intracellular signaling domain, wherein the polypeptides assemble together to form a multichain CAR. 
     The extracellular ligand-binding domain of a CAR specifically binds to a target of interest. The target of interest can be any molecule of interest, including, for example, without limitation any one or more of those presented in Table 11. 
     
       
         
           
               
             
               
                 TABLE 11 
               
               
                   
               
               
                 List of targets of interest 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
            
               
                 BCMA 
               
               
                 EGFRvIII 
               
               
                 Flt-3 
               
               
                 WT-1 
               
               
                 CD20 
               
               
                 CD23 
               
               
                 CD30 
               
               
                 CD38 
               
               
                 CD70 
               
               
                 CD33 
               
               
                 CD133 
               
               
                 MHC-WT1 
               
               
                 TSPAN10 
               
               
                 MHC-PRAME 
               
               
                 Liv1 
               
               
                 ADAM10 
               
               
                 CHRNA2 
               
               
                 LeY 
               
               
                 NKG2D 
               
               
                 CS1 
               
               
                 CD44v6 
               
               
                 ROR1 
               
               
                 CD19 
               
               
                 Claudin-18.2 (Claudin-18A2 or Claudin18 isoform 2) 
               
               
                 DLL3 (Delta-like protein 3,  Drosophila  Delta homolog 3, Delta3) 
               
               
                 Muc17 (Mucin17, Muc3, Muc3) 
               
               
                 FAP alpha (Fibroblast Activation Protein alpha) 
               
               
                 Ly6G6D (Lymphocyte antigen 6 complex locus protein G6d 
               
               
                 c6orf23 
               
               
                 G6D 
               
               
                 MEGT1 
               
               
                 NG25) 
               
               
                 RNF43 (E3 ubiquitin-protein ligase RNF43, RING finger protein 43) 
               
               
                 ErbB2 (HER2/neu) 
               
               
                 carcinoembryonic antigen (CEA) 
               
               
                 epithelial cell adhesion molecule (EpCAM) 
               
               
                 epidermal growth factor receptor (EGFR) 
               
               
                 CD40 
               
               
                 disialoganglioside GD2 
               
               
                 GD3 
               
               
                 C-type lectin-like molecule-1 (CLL-1) 
               
               
                 ductal-epithelial mucine 
               
               
                 gp36 
               
               
                 TAG-72 
               
               
                 glycosphingolipids 
               
               
                 glioma-associated antigen 
               
               
                 β-human chorionic gonadotropin 
               
               
                 alphafetoprotein (AFP) 
               
               
                 lectin-reactive AFP 
               
               
                 thyroglobulin 
               
               
                 RAGE-1 
               
               
                 MN-CA IX 
               
               
                 human telomerase reverse transcriptase 
               
               
                 RU1 
               
               
                 RU2 (AS) 
               
               
                 intestinal carboxyl esterase 
               
               
                 mut hsp70-2 
               
               
                 M-CSF 
               
               
                 prostase 
               
               
                 prostase specific antigen (PSA) 
               
               
                 PAP 
               
               
                 NY-ESO-1 
               
               
                 LAGA-1a 
               
               
                 p53 
               
               
                 prostein 
               
               
                 PSMA 
               
               
                 survivin and telomerase 
               
               
                 prostate-carcinoma tumor antigen-1 (PCTA-1) 
               
               
                 MAGE 
               
               
                 ELF2M 
               
               
                 neutrophil elastase 
               
               
                 ephrin B2 
               
               
                 CD22 
               
               
                 insulin growth factor (IGFl)-1 
               
               
                 IGF-II 
               
               
                 IGFI receptor 
               
               
                 mesothelin 
               
               
                 a major histocompatibility complex (MHC) molecule presenting a tumor- 
               
               
                 specific peptide epitope 
               
               
                 5T4 
               
               
                 O 1 
               
               
                 Nkp30 
               
               
                 tumor stromal antigens 
               
               
                 the extra domain A (EDA) and extra domain B (EDB) of fibronectin  
               
               
                 and the AI domain of tenascin-C (TnC AI) and fibroblast associated  
               
               
                 protein (fap) 
               
               
                 LRP6 
               
               
                 melamona-associated Chondroitin Sulfate Proteoglycan (MCSP) 
               
               
                 MARTI 
               
               
                 MUC1 
               
               
                 LMP2 
               
               
                 Idiotype 
               
               
                 NY-ESO-1 
               
               
                 Ras mutant 
               
               
                 gp100 
               
               
                 proteinase 3 
               
               
                 bcr-abl 
               
               
                 tyrosinase 
               
               
                 hTERT 
               
               
                 EphA2 
               
               
                 ML-TAP 
               
               
                 ERG 
               
               
                 NA17 
               
               
                 PAX3 
               
               
                 ALK 
               
               
                 Androgen receptor 
               
               
                 a lineage-specific or tissue specific antigen such as CD3 
               
               
                 CD4 
               
               
                 CD8 
               
               
                 CD24 
               
               
                 CD25 
               
               
                 CD34 
               
               
                 CD79 
               
               
                 CD116 
               
               
                 CD117 
               
               
                 CD135 
               
               
                 CD123 
               
               
                 CD138 
               
               
                 CTLA-4 
               
               
                 B7-1 (CD80) 
               
               
                 B7-2 (CD86) 
               
               
                 endoglin 
               
               
                 a major histocompatibility complex (MHC) molecule 
               
               
                 MUC16 
               
               
                 PSCA 
               
               
                 Trop2 
               
               
                 CD171 (L1CAM) 
               
               
                 CA9 
               
               
                 STEAP1 
               
               
                 VEGFR2 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the extracellular ligand-binding domain of a CAR comprises an scFv comprising the light chain variable (VL) region and the heavy chain variable (VH) region of a target antigen specific monoclonal antibody joined by a flexible linker. Single chain variable region fragments are made by linking light and/or heavy chain variable regions by using a short linking peptide (Bird et al., Science 242:423-426, 1988) (e.g. glycine-serine containing linkers). In general, linkers can be short, flexible polypeptides and are generally comprised of about 20 or fewer amino acid residues. Linkers can in turn be modified for additional functions, such as attachment of drugs or attachment to solid supports. The single chain variants can be produced either recombinantly or synthetically. For synthetic production of scFv, an automated synthesizer can be used. For recombinant production of scFv, a suitable plasmid containing polynucleotide that encodes the scFv can be introduced into a suitable host cell, either eukaryotic, such as yeast, plant, insect or mammalian cells, or prokaryotic, such as  E. coli . Polynucleotides encoding the scFv of interest can be made by routine manipulations such as ligation of polynucleotides. The resultant scFv can be isolated using standard protein purification techniques known in the art. 
     The intracellular signaling domain of a CAR according to the invention is responsible for intracellular signaling following the binding of extracellular ligand-binding domain to the target resulting in the activation of the immune cell and immune response (Signals 1 and/or 2). The intracellular signaling domain has the ability to activate at least one of the normal effector functions of the immune cell in which the CAR is expressed. For example, the effector function of a T cell can be a cytolytic activity or helper activity including the secretion of cytokines. 
     In some embodiments, an intracellular signaling domain for use in a CAR can be the cytoplasmic sequences of, for example without limitation, the T cell receptor and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability. Intracellular signaling domains comprise two distinct classes of cytoplasmic signaling sequences: those that initiate antigen-dependent primary activation, and those that act in an antigen-independent manner to provide a secondary or co-stimulatory signal. Primary cytoplasmic signaling sequences can comprise signaling motifs which are known as immunoreceptor tyrosine-based activation motifs of ITAMs. ITAMs are well defined signaling motifs found in the intracytoplasmic tail of a variety of receptors that serve as binding sites for syk/zap70 class tyrosine kinases. Examples of ITAM used in the invention can include as non-limiting examples those derived from TCRζ, FcRγ, FcRβ, FcRε, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b and CD66d. In some embodiments, the intracellular signaling domain of the CAR can comprise the CD3 signaling domain. In some embodiments the intracellular signaling domain of the CAR of the invention comprises a domain of a co-stimulatory molecule. 
     In some embodiments, the intracellular signaling domain of a CAR of the invention comprises a part of co-stimulatory molecule selected from the group consisting of fragment of 41BB (GenBank: AAA53133.) and CD28 (NP 006130.1). 
     CARs are expressed on the surface membrane of the cell. Thus, the CAR comprises a transmembrane domain. Suitable transmembrane domains for a CAR disclosed herein have the ability to (a) be expressed at the surface of a cell, preferably an immune cell such as, for example without limitation, lymphocyte cells or Natural killer (NK) cells, and (b) interact with the ligand-binding domain and intracellular signaling domain for directing cellular response of immune cell against a predefined target cell. The transmembrane domain can be derived either from a natural or from a synthetic source. The transmembrane domain can be derived from any membrane-bound or transmembrane protein. As non-limiting examples, the transmembrane polypeptide can be a subunit of the T cell receptor such as α, β, γ or δ, polypeptide constituting CD3 complex, IL-2 receptor p55 (a chain), p75 (β chain) or γ chain, subunit chain of Fc receptors, in particular Fcγ receptor III or CD proteins. Alternatively, the transmembrane domain can be synthetic and can comprise predominantly hydrophobic residues such as leucine and valine. In some embodiments said transmembrane domain is derived from the human CD8a chain (e.g., NP 001139345.1). The transmembrane domain can further comprise a stalk domain between the extracellular ligand-binding domain and said transmembrane domain. A stalk domain may comprise up to 300 amino acids, preferably 10 to 100 amino acids and most preferably 25 to 50 amino acids. Stalk region may be derived from all or part of naturally occurring molecules, such as from all or part of the extracellular region of CD8, CD4, or CD28, or from all or part of an antibody constant region. Alternatively the stalk domain may be a synthetic sequence that corresponds to a naturally occurring stalk sequence, or may be an entirely synthetic stalk sequence. In some embodiments said stalk domain is a part of human CD8a chain (e.g., NP 001139345.1). In another particular embodiment, said transmembrane and hinge domains comprise a part of human CD8a chain. In some embodiments, CARs disclosed herein can comprise an extracellular ligand-binding domain that specifically binds BCMA, CD8a human hinge and transmembrane domains, the CD3 signaling domain, and 4-1BB signaling domain. 
     In some embodiments, a CAR can be introduced into an immune cell as a transgene via a plasmid vector. In some embodiments, the plasmid vector can also contain, for example, a selection marker which provides for identification and/or selection of cells which received the vector. 
     Table 8 provides exemplary sequences of CAR components that can be used in the CARs disclosed herein. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Exemplary Sequences 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ ID 
               
               
                 Domain 
                 Amino acid sequence 
                 NO: 
               
               
                   
               
               
                 V5 epitope tag 
                 IPNPLLGLDST 
                 122 
               
               
                   
               
               
                 2173 scFv 
                 ETQLVQSGAEVKKPGESLRISCKGSGFNIEDYYIH 
                 123 
               
               
                   
                 WVRQMPGKGLEWMGRIDPENDETKYGPIFQGH 
                   
               
               
                   
                 VTISADTSINTVYLQWSSLKASDTAMYYCAFRG 
                   
               
               
                   
                 GVYWGQGTTVTVSSGGGGSGGGGSGGGGSGGG 
                   
               
               
                   
                 GSDVVMTQSPDSLAVSLGERATINCKSSQSLLDS 
                   
               
               
                   
                 DGKTYLNWLQQKPGQPPKRLISLVSKLDSGVPD 
                   
               
               
                   
                 RFSGSGSGTDFTLTISSLQAEDVAVYYCWQGTHF 
                   
               
               
                   
                 PGTFGGGTKVEIK 
                   
               
               
                   
               
               
                 CD8 hinge and 
                 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAV 
                 124 
               
               
                 transmembrane 
                 HTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC 
                   
               
               
                   
               
               
                 4-1BB intracellular 
                 KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPE 
                 125 
               
               
                 signaling 
                 EEEGGCEL 
                   
               
               
                   
               
               
                 CD3z intracellular 
                 RVKFSRSADAPAYQQGQNQLYNELNLGRREEYD 
                 126 
               
               
                 signaling 
                 VLDKRRGRDPEMGGKPRRKNPQEGLYNELQKD 
                   
               
               
                   
                 KMAEAYSEIGMKGERRRGKGHDGLYQGLSTAT 
                   
               
               
                   
                 KDTYDALHMQALPPR 
                   
               
               
                   
               
               
                 BFP (blue fluorescent 
                 MSELIKENMHMKLYMEGTVDNHHFKCTSEGEG 
                 127 
               
               
                 protein) 
                 KPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGS 
                   
               
               
                   
                 KTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG 
                   
               
               
                   
                 VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVM 
                   
               
               
                   
                 QKKTLGWEAFTETLYPADGGLEGRNDMALKLV 
                   
               
               
                   
                 GGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYR 
                   
               
               
                   
                 LERIKEANNETYVEQHEVAVARYCDLPSKLGHK 
                   
               
               
                   
                 LN 
                   
               
               
                   
               
               
                 P2A 
                 GSGATNFSLLKQAGDVEENPGP 
                 128 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the CAR-immune cell (e.g., CAR-T cell) of the disclosure comprises a polynucleotide encoding a suicide polypeptide, such as for example RQR8. See, e.g., WO2013153391A, which is hereby incorporated by reference in its entirety. In some embodiments, a suicide polypeptide is expressed on the surface of the cell. In some embodiments, a suicide polypeptide is included in the CAR construct. In some embodiments, a suicide polypeptide is not part of the CAR construct. 
     In some embodiments, the extracellular domain of any one of CARs disclosed herein may comprise one or more epitopes specific for (specifically recognized by) a monoclonal antibody. These epitopes are also referred to herein as mAb-specific epitopes. Exemplary mAb-specific epitopes are disclosed in International Patent Publication No. WO 2016/120216, which is incorporated herein in its entirety. In these embodiments, the extracellular domain of the CARs comprise antigen binding domains that specifically bind to a target of interest and one or more epitopes that bind to one or more monoclonal antibodies (mAbs). CARs comprising the mAb-specific epitopes can be single-chain or multi-chain. 
     The inclusion of epitopes specific for monoclonal antibodies in the extracellular domain of the CARs described herein allows sorting and depletion of engineered immune cells expressing the CARs. In some embodiments, allowing for depletion provides a safety switch in case of deleterious effects, e.g., upon administration to a subject. 
     Methods of preparing immune cells for use in immunotherapy are also provided herein. In some embodiments, the methods comprise introducing a PD-1 chimeric cytokine receptor and a CAR into immune cells, and expanding the cells. In some embodiments, the invention relates to a method of engineering an immune cell comprising: providing a cell and expressing a PD-1 chimeric cytokine receptor, and expressing at the surface of the cell at least one CAR. In some embodiments, the method comprises: transfecting the cell with at least one polynucleotide encoding a PD-1 chimeric cytokine receptor, and at least one polynucleotide encoding a CAR, and expressing the polynucleotides in the cell. In some embodiments, the method comprises: transfecting the cell with at least one polynucleotide encoding a PD-1 chimeric cytokine receptor, at least one polynucleotide encoding a CAR, and expressing the polynucleotides in the cell. 
     In some embodiments, the polynucleotides encoding the PD-1 chimeric cytokine receptor and CAR are present in one or more expression vectors for stable expression in the cells. In some embodiments, the polynucleotides are present in viral vectors for stable expression in the cells. In some embodiments, the viral vectors may be for example, lentiviral vectors or adenoviral vectors. 
     In some embodiments, polynucleotides encoding polypeptides according to the present disclosure can be mRNA which is introduced directly into the cells, for example by electroporation. In some embodiments, CytoPulse electroporation technology, such as PulseAgile, can be used to transiently permeabilize living cells for delivery of material into the cells (e.g. U.S. Pat. No. 6,078,490; PCT/US2011/000827; and PCT/US2004/005237). Parameters can be modified in order to determine conditions for high transfection efficiency with minimal mortality. 
     Also provided herein are methods of transfecting an immune cell, e.g a T-cell. In some embodiments, the method comprises: contacting a T-cell with RNA and applying to the T-cell an agile pulse sequence. In some embodiments, a method of transfecting an immune cell (e.g. T-cell) comprising contacting the immune cell with RNA and applying to the cell an agile pulse sequence. 
     In some embodiments, the method can further comprise a step of genetically modifying a cell by inactivating at least one gene expressing, for example without limitation, a component of the TCR, a target for an immunosuppressive agent, an HLA gene, and/or an immune checkpoint protein such as, for example, PDCD1 or CTLA-4. By inactivating a gene it is intended that the gene of interest is not expressed in a functional protein form. In some embodiments, the gene to be inactivated is selected from the group consisting of, for example without limitation, TCRα, TCRβ, CD52, GR, deoxycytidine kinase (DCK), PD-1, and CTLA-4. In some embodiments the method comprises inactivating one or more genes by introducing into the cells a rare-cutting endonuclease able to selectively inactivate a gene by selective DNA cleavage. In some embodiments the rare-cutting endonuclease can be, for example, a transcription activator-like effector nuclease (TALE-nuclease) or CRISPR-based endonuclease (e.g Cas-9 or Cas12a). 
     In another aspect, a step of genetically modifying cells can comprise: modifying immune cells (e.g. T-cells) by inactivating at least one gene expressing a target for an immunosuppressive agent, and; expanding the cells, optionally in presence of the immunosuppressive agent. 
     In some embodiments, the engineered immune cells (e.g. T-cells) provided herein exhibit improved cytotoxicity, increased expansion, and/or increased levels of memory phenotype markers upon contact with a PD-L1 or PD-L2 ligand or PD-1 antibody that binds to the ectodomain of the PD-1 chimeric cytokine receptor relative to engineered immune cells that do not express the PD-1 chimeric cytokine receptor. 
     In some embodiments, the engineered immune cells (e.g. T-cells) provided herein exhibit (i) increased in vivo persistence, (ii) increased STAT activation, (iii) increased cytotoxicity, (iv) increased levels of memory phenotype markers, (v) increased expansion (proliferation), or combinations of these functional features, upon contact with a PD-L1 or PD-L2 ligand or PD-1 antibody that binds to the ectodomain of the PD-1 chimeric cytokine receptor relative to engineered immune cells that do not express the PD-1 chimeric cytokine receptor. In some embodiments, the improvement in the one or more functional features described herein is dose-dependent, i.e., the functional activity of the immune cell comprising the PD-1 chimeric cytokine receptors increases upon contact with increasing doses of the PD-L1/PD-L2/PD-1 antibody. In some embodiments, STAT5 activated by the engineered immune cell comprising one or more PD-1 chimeric cytokine receptors disclosed are STAT1, STAT2, STAT3, STAT4, STAT5, STAT6, or combinations thereof. In one embodiment, memory phenotype markers increased or maintained by the immune cell comprising the inducible chimeric cytokine receptor include stem cell memory (Tscm) marker and central memory (Tcm) marker. 
     In some embodiments, the improvement in one or more functional features exhibited by an engineered immune cell comprising an inducible chimeric cytokine receptor provided herein is at least about 2 fold, 2.5 fold, 3 fold, 3.5 fold, 4 fold, 4.5 fold, 5 fold, 6 fold, 7 fold, 8 fold, 9 fold, 10 fold, 15 fold, 20 fold, 25 fold, 30 fold, 40 fold, 50 fold, 60 fold, 70 fold, 80 fold, 90 fold, 100 fold, 125 fold, 150 fold, 200 fold, 250 fold, 300 fold, 350 fold, 400 fold, 450 fold, or even about 10500 fold, including values and ranges therebetween, compared to an immune cell that does not express the PD-1 chimeric cytokine receptor. 
     In some embodiments, the improvement in one or more functional features exhibited by an engineered immune cell comprising a PD-1 chimeric cytokine receptor provided herein is at least about 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150%, 200%, 250%, 300%, 350%, 400%, or even about 80%500%, including values and ranges therebetween, compared to an engineered immune cell that does not express the PD-1 chimeric cytokine receptor. 
     III. Therapeutic Methods 
     Provided herein are pharmaceutical compositions comprising cells bearing the chimeric cytokine receptors and CARs of the disclosure. 
     Engineered PD-1 chimeric cytokine receptor-bearing and CAR-bearing immune cells (e.g. T-cells) obtained by the methods described above, or cell lines derived from such engineered immune cells, can be used as a medicament. In some embodiments, such a medicament can be used for treating a disorder such as for example a viral disease, a bacterial disease, a cancer, an inflammatory disease, an immune disease, or an aging-associated disease. In some embodiments, the cancer is a solid cancer. In some embodiments the cancer is a liquid cancer. The cancer can be selected from the group consisting of gastric cancer, sarcoma, lymphoma, leukemia, head and neck cancer, thymic cancer, epithelial cancer, salivary cancer, liver cancer, stomach cancer, thyroid cancer, lung cancer, ovarian cancer, breast cancer, prostate cancer, esophageal cancer, pancreatic cancer, glioma, leukemia, multiple myeloma, renal cell carcinoma, bladder cancer, cervical cancer, choriocarcinoma, colon cancer, oral cancer, skin cancer, and melanoma. In some embodiments, the subject is a previously treated adult subject with locally advanced or metastatic melanoma, squamous cell head and neck cancer (SCHNC), ovarian carcinoma, sarcoma, or relapsed or refractory classic Hodgkin&#39;s Lymphoma (cHL). 
     In some embodiments, engineered immune cells, or cell line derived from the engineered immune cells, can be used in the manufacture of a medicament for treatment of a disorder in a subject in need thereof. In some embodiments, the disorder can be, for example, a cancer, an autoimmune disorder, or an infection. 
     Also provided herein are methods for treating subjects in need of such treatment. 
     As used herein, the term “subject” refers to any vertebrate including, without limitation, humans and other primates (e.g., chimpanzees, cynomologous monkeys, and other apes and monkey species), farm animals (e.g., cattle, sheep, pigs, goats and horses), domestic mammals (e.g., dogs and cats), laboratory animals (e.g., rabbits, rodents such as mice, rats, and guinea pigs), and birds (e.g., domestic, wild and game birds such as chickens, turkeys and other gallinaceous birds, ducks, geese, and the like). In some embodiments, the subject is a mammal. In exemplary embodiments, the subject is a human. 
     In some embodiments the method comprises providing immune cells of the disclosure, bearing the PD-1 chimeric cytokine receptors and CARs described herein to a subject in need thereof. 
     In some embodiments, PD-1 chimeric cytokine receptor and CAR-bearing T-cells of the invention can undergo robust in vivo T-cell expansion and can persist for an extended amount of time. 
     Methods of treatment of the invention can be ameliorating, curative or prophylactic. The method of the invention may be either part of an autologous immunotherapy or part of an allogenic immunotherapy treatment. 
     In another aspect, the invention provides a method of inhibiting tumor growth or progression in a subject who has a tumor, comprising administering to the subject an effective amount of PD-1 chimeric cytokine receptor-expressing and CAR-expressing immune cells as described herein. In another aspect, the invention provides a method of inhibiting or preventing metastasis of cancer cells in a subject, comprising administering to the subject in need thereof an effective amount of engineered immune cells as described herein. In another aspect, the invention provides a method of inducing tumor regression in a subject who has a tumor, comprising administering to the subject an effective amount of engineered immune cells as described herein. 
     In some embodiments, the engineered T-cells herein can be administered parenterally in a subject. 
     Also provided is the use of any of the engineered T-cells provided herein in the manufacture of a medicament for the treatment of cancer or for inhibiting tumor growth or progression in a subject in need thereof. 
     In some embodiments, treatment can be administrated into subjects undergoing an immunosuppressive treatment. Indeed, the invention preferably relies on cells or population of cells, which have been made resistant to at least one immunosuppressive agent due to the inactivation of a gene encoding a receptor for such immunosuppressive agent. In this aspect, the immunosuppressive treatment should help the selection and expansion of the T-cells according to the invention within the subject. The administration of the cells or population of cells according to the invention may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. The compositions described herein may be administered to a subject subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous or intralymphatic injection, or intraperitoneally. Cells bearing the CARs of the disclosure or the pharmaceutical compositions thereof may be administered via one or more of the following routes of administration: intravenous, intraocular, intravitreal, intramuscular, subcutaneous, topical, oral, transdermal, intraperitoneal, intraorbital, by implantation, by inhalation, intrathecal, intraventricular, via the ear, or intranasal. 
     In some embodiments the administration of the cells or population of cells (bearing the chimeric cytokine receptors and CARs of the disclosure) can comprise administration of, for example, about 10 4  to about 10 9  cells per kg body weight including all integer values of cell numbers within those ranges. In some embodiments the administration of the cells or population of cells can comprise administration of about 10 4  to 10 5  cells per kg body weight, 10 5  to 10 6  cells per kg body weight, 10 6  to 10 7  cells per kg body weight, 10 7  to 10 8  cells per kg body weight, or 10 8  to 10 9  cells per kg body weight. The cells or population of cells can be administrated in one or more doses. In some embodiments, said effective amount of cells can be administrated as a single dose. In some embodiments, said effective amount of cells can be administrated as more than one dose over a period time. Timing of administration is within the judgment of managing physician and depends on the clinical condition of the subject. The cells or population of cells may be obtained from any source, such as a blood bank or a donor. While individual needs vary, determination of optimal ranges of effective amounts of a given cell type for a particular disease or conditions within the skill of the art. An effective amount means an amount which provides a therapeutic or prophylactic benefit. The dosage administrated will be dependent upon the age, health and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired. In some embodiments, an effective amount of cells or composition comprising those cells are administrated parenterally. In some embodiments, administration can be an intravenous administration. In some embodiments, administration can be directly done by injection within a tumor. 
     The methods can further comprise administering one or more agents to a subject prior to administering the engineered immune cells bearing a CAR and a PD-1 chimeric cytokine receptor provided herein. In certain embodiments, the agent is a lymphodepleting (preconditioning) regimen. For example, methods of lymphodepleting a subject in need of such therapy comprise administering to the subject specified beneficial doses of cyclophosphamide (between 200 mg/m 2 /day and 2000 mg/m 2 /day, about 100 mg/m 2 /day and about 2000 mg/m 2 /day; e.g., about 100 mg/m 2 /day, about 200 mg/m 2 /day, about 300 mg/m 2 /day, about 400 mg/m 2 /day, about 500 mg/m 2 /day, about 600 mg/m 2 /day, about 700 mg/m 2 /day, about 800 mg/m 2 /day, about 900 mg/m 2 /day, about 1000 mg/m 2 /day, about 1500 mg/m 2 /day or about 2000 mg/m 2 /day) and specified doses of fludarabine (between 20 mg/m 2 /day and 900 mg/m 2 /day, between about 10 mg/m 2 /day and about 900 mg/m 2 /day; e.g., about 10 mg/m 2 /day, about 20 mg/m 2 /day, about 30 mg/m 2 /day, about 40 mg/m 2 /day, about 40 mg/m 2 /day, about 50 mg/m 2 /day, about 60 mg/m 2 /day, about 70 mg/m 2 /day, about 80 mg/m 2 /day, about 90 mg/m 2 /day, about 100 mg/m 2 /day, about 500 mg/m 2 /day or about 900 mg/m 2 /day). An exemplary dosing regimen involves treating a subject comprising administering daily to the patient about 300 mg/m 2 /day of cyclophosphamide in combination or before or after administering about 30 mg/m 2 /day of fludarabine for three days prior to administration of a therapeutically effective amount of engineered immune cells to the patient. 
     In some embodiments, notably in the case when the engineered cells provided herein have been gene edited to eliminate or minimize surface expression of CD52, lymphodepletion further comprises administration of an anti-CD52 antibody, such as alemtuzumab. In some embodiments, the CD52 antibody is administered at a dose of about 1-20 mg/day IV, e.g., about 13 mg/day IV for 1, 2, 3 or more days. The antibody can be administered in combination with, before, or after administration of other elements of a lymphodepletion regime (e.g., cyclophosphamide and/or fludarabine). 
     In certain embodiments, compositions comprising CAR-expressing immune effector cells disclosed herein may be administered in conjunction with any number of chemotherapeutic agents. 
     IV. Kits and Articles of Manufacture 
     The present disclosure provides kits comprising any one or more of the PD-1 chimeric cytokine receptors and PD-1 chimeric cytokine receptors-bearing cells described herein, and pharmaceutical compositions thereof. The present disclosure also provides articles of manufacture comprising any one or more of the PD-1 chimeric cytokine receptors and PD-1 chimeric cytokine receptors-bearing CAR-I cells described herein, pharmaceutical compositions thereof, and kits described herein. 
     The following examples are included for illustrative purposes and are not intend to limit the scope of the disclosure. 
     All patent and non-patent documents referenced throughout this disclosure are incorporated by reference herein in their entirety for all purposes. 
     EXAMPLES 
     In referring to the chimeric cytokine receptors of the disclosure it is noted that certain figures refer to a “switch.” As used herein, a “switch” is used interchangeably with a “chimeric cytokine receptor”. 
     The following notations are used throughout the drawings and examples. 
     FKBP switch=a FKBP chimeric cytokine receptor 
     WT PD1 switch=a chimeric cytokine receptor with a wild type PD-1 ectodomain 
     2×PD1 switch=a chimeric cytokine receptor with 2 PD-1 ectodomains in tandem 
     3×PD1 switch=a chimeric cytokine receptor with 3 PD-1 ectodomains in tandem 
     HA PD1 switch=a chimeric cytokine receptor with a high affinity PD-1 ecotodomain 
     HA PD1 DN=a chimeric cytokine receptor with a dominant negative high affinity PD-1 domain 
     Example 1: Coupling of PD-1 Engagement with Cytokine Signaling 
     To couple simultaneous PD-1 engagement with cytokine signaling, a PD-1 chimeric cytokine receptor was constructed, composed of the following modules: (i) the wild type (WT) PD-1 ectodomain (21-207), (ii) a transmembrane domain with an intracellular portion having a JAK2-activating domain and (iii) STAT-recruiting domains comprising Stat-recruiting (Stat-activating) domains from cytokine receptor tails (cytotails). With this design, it was determined whether clustering or dimerization of the PD-1 chimeric cytokine receptor, in response to PD-1 ligands or anti-PD-1 antibodies, respectively, could activate the JAK-Stat pathway and mimic cytokine signaling. In order to minimize the transgene payload in the lentiviral vector, JAK2-activating domains derived from natural cytokine receptors known to signal as homodimers, such as TpoR (TPOR/MPLR(478-582)), GCSFR (GCSFR(614-710)) and GP130 (GP130(609-700)), were selected for initial evaluation. 
     Two Stat-activating cytoplasmic domains derived from IL7R(316-459) and IL12Rb(775-825) were linked in tandem mimic signaling in response to IL7 and IL12, respectively. To demonstrate the utility of the PD-1 chimeric cytokine receptor in the context of CAR-T-cells, the PD-1 chimeric cytokine receptor was cloned into a lentiviral vector encoding a second generation EGFRvIII-specific CAR (2173scFv; described in Sci Transl Med. 2015 Feb. 18; 7(275): 275ra22.); furthermore, to permit stoichiometric co-expression of the PD-1 chimeric cytokine receptor and the CAR, both genes were linked via a P2A peptide linker. To facilitate the detection of transduced cells, a v5 epitope tag (KPIPNPLLGLDST (SEQ ID NO: 167)) was inserted between the scFv and CD8 hinge domain of the CAR. A HEK293T-cell reporter assay was used to test the inducibility and magnitude of cytokine signaling. Briefly, 20,000 HEK293T-cells were plated into each well of a poly-L-lysine-coated 96-well flat-bottom plate and allowed to adhere overnight. A PD-1 chimeric cytokine receptor (2.5 ng), a Stat response element that drives Firefly Luciferase (100 ng; Promega) and Renilla Luciferase control reporter vector (1 ng; Promega) were mixed in a final volume of 5 uL in Opti-MEM (Gibco) (“DNA mix”). As a positive control, a FKBP chimeric cytokine receptor was used, in which the WT PD-1 ectodomain was replaced with FKBP12(F36V), so that cytokine signaling could be inducible by the small molecule, AP1903. 0.3 uL Lipofectamine 2000 (Invitrogen) in 5 uL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 10 uL was added to each well containing HEK-293T. 24 hours after transfection, cells were either left untreated, or treated with the indicated concentrations of anti-PD-1 (Nivolumab; Selleck Chemical), recombinant human PD-L1-Fc (Biolegend), anti-Human IgG Fc gamma secondary antibody (Thermo Fisher), or AP1903 (Apex Bio). As negative controls, hIgG4 and hIgG1 isotype controls were added in place of anti-PD-1 and PD-L1-Fc, respectively. All treatments were diluted in serum-free media. 24 hours after treatment, Stat reporter activity was evaluated using the Dual-Glo Luciferase Assay System (Promega). Fold induction of Stat5 reporter activity was normalized to that of HEK293T-cells transfected with all vectors except for the chimeric cytokine receptor and that were left untreated. 
       FIGS. 1A-C  show schematics of the various embodiments of the PD-1 chimeric cytokine receptor. 
       FIG. 1B  shows a schematic of the PD-1 chimeric cytokine receptor CAR vector.  FIG. 1D  shows a schematic of the PD-1 chimeric cytokine receptor CAR vector, and the positive control FKBP chimeric cytokine receptor CAR vector. 
       FIGS. 2A-D  show Stat reporter activity in response to PD-L1-Fc ( FIGS. 2A-B ) and anti-PD-1 ( FIGS. 2C-D ).  FIGS. 2A-B  show that as compared to their untreated counterparts, PD-1 chimeric cytokine receptors induced weak Stat reporter activity in response to high concentrations of crosslinked PD-L1 (i.e. hPD-L1-Fc (10 ug/ml)+2Ab (25 ug/ml)).  FIGS. 2C-D  show that as compared to their untreated counterparts, PD-1 chimeric cytokine receptors induced weak Stat reporter activity in response to high concentrations of crosslinked anti-PD-1 (i.e. aPD-1 (10 ug/ml)+2Ab (25 ug/ml)), with the TpoR(478-582) TM/JAK variant showing the best response. Although the FKBP chimeric cytokine receptor positive control showed basal activity in the absence of AP1903, it induced high Stat reporter activity in response to AP1903. 
     Similar experiments were then carried out in the context of primary human CAR-T-cells. To make lentivirus encoding PD-1 chimeric cytokine receptor CARs, HEK293T-cells were plated at 0.45 million cells per mL in 2 mL of DMEM (Gibco) supplemented with 10% FBS (Hyclone) per well of a 6-well plate on Day −1. On Day 0, the lentivirus was prepared by mixing together lentiviral packaging vectors 1.5 ug psPAX2, 0.5 ug pMD2G, and 0.5 ug of the appropriate transfer CAR vector in 250 uL Opti-MEM (Gibco) per well of the 6-well plate (“DNA mix”). 10 uL Lipofectamine 2000 (Invitrogen) in 250 uL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 500 uL was slowly added to the sides of the wells containing HEK293T. On Day 0, purified T-cells were activated in X-Vivo-15 medium (Lonza) supplemented with 100 IU/mL human IL-2 (Miltenyi Biotec), 10% FBS (Hyclone), and human T TransAct (Miltenyi Biotec, Cat #130-111-160, 1:100 dilution) in a Grex-24 plate (Wilson Wolf, cat #80192M). On Day 1, the media from each well of HEK293T-cells in the 6-well plate was replaced with 2 mL per well of T-cell transduction media, i.e., X-Vivo-15 supplemented with 10% FBS. On Day 2, T-cells were resuspended at 0.5 million cells per mL in 1 mL of T-cell transduction media per well of a Grex-24 plate. The lentiviral supernatants from HEK293T-cells were harvested and passed through a 0.45 micron filter (EMD Millipore) to remove cell debris, and then added to the T-cells along with 100 IU/mL human IL-2. On Day 5, 4.5 mL of T-cell expansion media, i.e., X-Vivo-15 supplemented with 5% human AB serum (Gemini Bio) and 100 IU/mL human IL-2 was added to each well of a Grex-24 plate. Cells were expanded into larger G-Rex vessels (Wilson Wolf) as needed using T-cell expansion media. On Day 13 or 14, transduction efficiency and PD-1 chimeric cytokine receptor expression was determined by detecting the percentage of T-cells that bound a FITC-conjugated v5 tag monoclonal antibody (Thermo Fisher) and a PE/Cy7-conjugated anti-human PD-1 (Biolegend) using flow cytometry. On Day 14 or 15, the CAR-T-cell products were cryopreserved and thawed as needed for further assays. 
       FIG. 3  shows CAR and PD-1 chimeric cytokine receptor expression on the Day 14 CAR-T-cell product. As compared to FKBP chimeric cytokine receptor CAR-T-cells on which little to no PD-1 was detected, high levels of PD-1 was detected on PD-1 chimeric cytokine receptor CAR-T-cells, indicating that the CAR and PD-1 chimeric cytokine receptor were coexpressed. 
     To determine the inducibility and magnitude of cytokine signaling in PD-1 chimeric cytokine receptor CAR-T-cells, the thawed CAR-T-cell product was serum starved in 100 uL serum-free RPMI (Corning) for 4 hours in humidified incubator at 37° C. with 5% CO 2 , then treated with anti-PD-1 (10 ug/mL Nivolumab; Selleck Chemical), recombinant human PD-L1-Fc (10 ug/mL; Biolegend), anti-Human IgG Fc gamma secondary antibody (25 ug/mL; Thermo Fisher) for 1 hour. 40 minutes into the treatment, an antibody cocktail comprising BUV395-conjugated anti-human CD3 (Biolegend) and FITC-conjugated v5 tag monoclonal antibody (Thermo Fisher) were added to the cells and allowed to incubate for the final 20 minutes. After 1 hour of treatment, cells were fixed by the addition of 35 uL of 16% paraformaldehyde was added to each 100 uL sample and allowed to incubate for 15 minutes at 37° C. Cells were then washed three times with PBS, and permeabilized in 100% cold methanol for 1 or 2 nights at −20° C. On the day of FACS analysis, cells were washed three times with PBS, Fc-blocked, and stained with AlexaFluor647-conjugated anti-mouse/human Stat5 (pY694) (BD Biosciences) diluted in PBS+1% BSA. After a 1 hour incubation at room temperature in the dark, cells were washed three times before FACS analysis. 
       FIGS. 4A-B  show phospho-Stat5 staining in human CAR-T-cells following the treatment with PD-L1-Fc ( FIG. 4A ) or anti-PD-1 ( FIG. 4B ). PD-1 chimeric cytokine receptor CAR-T-cells bearing the GCSFR(614-710) TM/JAK2-activating domain showed high basal activity in the absence of treatment, and no induction of pStat5 in the presence of treatment. PD-1 chimeric cytokine receptor CAR-T-cells bearing the GP130(609-700) TM/JAK2-activating domain showed the lowest basal activity in the absence of treatment, no pStat5 induction in response to PD-L1 or anti-PD-1, and moderate pStat5 induction in response to crosslinked anti-PD-1. PD-1 chimeric cytokine receptor CAR-T-cells bearing the TpoR(478-582) TM/JAK2-activating domain showed the moderate basal activity in the absence of treatment, no pStat5 induction in response to PD-L1 or anti-PD-1, and strong pStat5 induction in response to crosslinked anti-PD-1. 
     Example 2: Optimization of the PD-1 Chimeric Cytokine Receptor 
     Compared to the high affinity interaction between Nivolumab and PD-1 (&lt;10 pM), PD-1 binds to PD-L1 and PD-L2 through a low-affinity (1-10 uM), monomeric interaction. The observation that crosslinked anti-PD-1 (Nivolumab) was able to activate the PD-1 chimeric cytokine receptor bearing the TpoR(478-582) TM/JAK2-activating domain indicating that PD-1 chimeric cytokine receptor activation could be made more efficient by enhancing its affinity or avidity of interaction with its inducer (PD-1 ligands or anti-PD-1). To increase the avidity of the PD-1 chimeric cytokine receptor, multiple WT PD-1 ectodomains were engineered in tandem. To increase the affinity of the PD-1 chimeric cytokine receptor, mutations to the PD-1 ectodomains were introduced, aimed at increasing binding affinity to its ligands (henceforth referred to as “high affinity (HA) PD-1”). 
     To demonstrate the utility of these PD-1 chimeric cytokine receptors in the context of CAR-T-cells, each PD-1 chimeric cytokine receptor was cloned into a lentiviral vector encoding a second generation EGFRvIII-specific CAR (2173scFv; described in Sci Transl Med. 2015 Feb. 18; 7(275): 275ra22.); furthermore, to permit stoichiometric co-expression of the PD-1 chimeric cytokine receptor and the CAR, both genes were linked via a P2A peptide linker. To facilitate the detection of transduced cells, a v5 epitope tag (KPIPNPLLGLDST (SEQ ID NO: 167)) was inserted between the scFv and CD8 hinge domain. 
       FIGS. 5A-D  show a schematic of PD-1 chimeric cytokine receptors with various ectodomains engineered for enhanced avidity or affinity.  FIG. 5A  shows the prototypic WT PD-1 chimeric cytokine receptor bearing the TpoR(478-582) TM/JAK2-activating domain;  FIG. 5B  shows the 2×PD-1 chimeric cytokine receptor bearing two WT PD-1 ectodomains in tandem;  FIG. 5C  shows the 3×PD-1 chimeric cytokine receptor bearing three WT PD-1 ectodomains in tandem;  FIG. 5D  shows the high affinity (HA) PD-1 chimeric cytokine receptor with ectodomain mutations. 
     A HEK293T-cell reporter assay was used to test the inducibility and magnitude of cytokine signaling. Briefly, 20,000 HEK293T-cells were plated into each well of a poly-L-lysine-coated 96-well flat-bottom plate and allowed to adhere overnight. A PD-1 chimeric cytokine receptor (2.5 ng), a Stat response element that drives Firefly Luciferase (100 ng; Promega) and Renilla Luciferase control reporter vector (1 ng; Promega) were mixed in a final volume of 5 uL in Opti-MEM (Gibco) (“DNA mix”). As a positive control, a FKBP chimeric cytokine receptor was used, in which the WT PD-1 ectodomain was replaced with FKBP12(F36V), so that cytokine signaling could be inducible by the small molecule, AP1903. 0.3 uL Lipofectamine 2000 (Invitrogen) in 5 uL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 10 uL was added to each well containing HEK-293T. 24 hours after transfection, cells were either left untreated, or treated with the indicated concentrations of anti-PD-1 (Nivolumab; Selleck Chemical), recombinant human PD-L1-Fc (Biolegend), anti-Human IgG Fc gamma secondary antibody (Thermo Fisher), or AP1903 (Apex Bio). As negative controls, hIgG4 and hIgG1 isotype controls were added in place of anti-PD-1 and PD-L1-Fc, respectively. All treatments were diluted in serum-free media. 24 hours after treatment, Stat reporter activity was evaluated using the Dual-Glo Luciferase Assay System (Promega). Fold induction of Stat5 reporter activity was normalized to that of HEK293T-cells transfected with all vectors except for the chimeric cytokine receptor receptor and that were left untreated. 
       FIGS. 6A-D  show the response of PD-1 chimeric cytokine receptors bearing ectodomain modifications following treatment with PD-L1-Fc ( FIGS. 6A-B ) and anti-PD-1 ( FIGS. 6C-D ).  FIGS. 6A-B  show that compared to its untreated counterpart and to the WT PD-1 chimeric cytokine receptor, the HA PD-1 chimeric cytokine receptor permitted enhanced Stat5 ( FIG. 6A ) and Stat4 ( FIG. 6B ) reporter activity in response to high concentrations of crosslinked PD-L1-Fc; whereas the 2×PD-1 chimeric cytokine receptor permitted enhanced Stat5, but not Stat4, reporter activity.  FIGS. 6C-d  show that compared to its untreated counterpart and to the WT PD-1 chimeric cytokine receptor, the HA PD-1 chimeric cytokine receptor permitted enhanced Stat5 ( FIG. 6C ) and Stat4 ( FIG. 6D ) reporter activity in response to high concentrations of crosslinked anti-PD-1; whereas the 2×PD-1 chimeric cytokine receptor permitted enhanced Stat5, but not Stat4, reporter activity. These indicated that enhancing the avidity of the ectodomain via tandem PD-1, or enhancing the affinity of the ectodomain via mutations, increase the responsiveness of the PD-1 chimeric cytokine receptor. Furthermore, the fact that the HA PD-1 chimeric cytokine receptor may be activated by anti-PD-1 indicates that the mutant HA ectodomain retains the ability to bind anti-PD-1 (i.e. Nivolumab). 
     Similar experiments were then carried out in the context of primary human CAR-T-cells. To make lentivirus encoding PD-1 chimeric cytokine receptor CARs, HEK293T-cells were plated at 0.45 million cells per mL in 2 mL of DMEM (Gibco) supplemented with 10% FBS (Hyclone) per well of a 6-well plate on Day −1. On Day 0, the lentivirus was prepared by mixing together lentiviral packaging vectors 1.5 ug psPAX2, 0.5 ug pMD2G, and 0.5 ug of the appropriate transfer CAR vector in 250 uL Opti-MEM (Gibco) per well of the 6-well plate (“DNA mix”). 10 uL Lipofectamine 2000 (Invitrogen) in 250 uL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 500 uL was slowly added to the sides of the wells containing HEK293T. On Day 0, purified T-cells were activated in X-Vivo-15 medium (Lonza) supplemented with 100 IU/mL human IL-2 (Miltenyi Biotec), 10% FBS (Hyclone), and human T TransAct (Miltenyi Biotec, Cat #130-111-160, 1:100 dilution) in a Grex-24 plate (Wilson Wolf, cat #80192M). On Day 1, the media from each well of HEK293T-cells in the 6-well plate was replaced with 2 mL per well of T-cell transduction media, i.e., X-Vivo-15 supplemented with 10% FBS. On Day 2, T-cells were resuspended at 0.5 million cells per mL in 1 mL of T-cell transduction media per well of a Grex-24 plate. The lentiviral supernatants from HEK293T-cells were harvested and passed through a 0.45 micron filter (EMD Millipore) to remove cell debris, and then added to the T-cells along with 100 IU/mL human IL-2. On Day 5, 4.5 mL of T-cell expansion media, i.e., X-Vivo-15 supplemented with 5% human AB serum (Gemini Bio) and 100 IU/mL human IL-2 was added to each well of a Grex-24 plate. Cells were expanded into larger G-Rex vessels (Wilson Wolf) as needed using T-cell expansion media. On Day 13 or 14, transduction efficiency and PD-1 chimeric cytokine receptor expression was determined by detecting the percentage of T-cells that bound a FITC-conjugated v5 tag monoclonal antibody (Thermo Fisher) using flow cytometry. On Day 14 or 15, the CAR-T-cell products were cryopreserved and thawed as needed for further assays. 
     To determine if the HA PD-1 ectodomain indeed enhances binding affinity to PD-L1, while retaining interactions with clinically-approved anti-PD-1 antibodies, the ability of HA PD-1 chimeric cytokine receptor CAR-T-cells to bind Nivolumab and Pembrolizumab was tested in a cell-binding assay. Briefly, WT PD-1 chimeric cytokine receptor CAR-T-cells or HA PD-1 chimeric cytokine receptor CAR-T-cells were Fc-blocked, and then incubated with the indicated concentrations of anti-PD-1 (i.e. Nivolumab or Pembrolizumab) diluted in PBS+1% BSA. After a 20 minute incubation at 4° C., cells were washed in PBS, and incubated with a PE-conjugated anti-human IgG Fc secondary antibody (Biolegend) diluted in PBS+1% BSA. After a 20 minute incubation at 4° C., cells were washed in PBS and analyzed by flow cytometry. 
       FIGS. 7A-B  show results from the cell binding assay between PD-1 chimeric cytokine receptor CAR-T-cells and PD-L1-Fc ( FIG. 7A ) or clinically-approved anti-PD-1 antibodies ( FIG. 7B ).  FIG. 7A  shows that while the WT PD-1 chimeric cytokine receptor did not bind to PD-L1-Fc at the concentrations tested, the HA PD-1 chimeric cytokine receptor dramatically improved binding to PD-L1-Fc.  FIG. 7B  shows that modification to the HA PD-1 ectodomain improved maximal binding (C max ) to Pembrolizumab, and improved both maximal binding (C max ) and sensitivity (EC 50 ) to Nivolumab. The HA PD-1 ectodomain variant improves responsiveness to both PD-L1 and clinically-approved anti-PD-1 antibodies. 
     To analyze the inducibility and magnitude of cytokine signaling in CAR-T-cells bearing these PD-1 chimeric cytokine receptor ectodomain variants, the thawed CAR-T-cell product was serum starved in 100 uL serum-free RPMI (Corning) for 4 hours in humidified incubator at 37° C. with 5% CO 2 , then treated with anti-PD-1 (10 ug/mL Nivolumab; Selleck Chemical), recombinant human PD-L1-Fc (10 ug/mL; Biolegend), anti-Human IgG Fc gamma secondary antibody (25 ug/mL; Thermo Fisher) for 1 hour. 40 minutes into the treatment, an antibody cocktail comprising BUV395-conjugated anti-human CD3 (Biolegend) and FITC-conjugated v5 tag monoclonal antibody (Thermo Fisher) were added to the cells and allowed to incubate for the final 20 minutes. After 1 hour of treatment, cells were fixed by the addition of 35 uL of 16% paraformaldehyde was added to each 100 uL sample and allowed to incubate for 15 minutes at 37° C. Cells were then washed three times with PBS, and permeabilized in 100% cold methanol for 1 or 2 nights at −20° C. On the day of FACS analysis, cells were washed three times with PBS, Fc-blocked, and stained with AlexaFluor647-conjugated anti-mouse/human Stat5 (pY694) (BD Biosciences) diluted in PBS+1% BSA. After a 1 hour incubation at room temperature in the dark, cells were washed three times before FACS analysis. 
       FIGS. 8A-B  show the response of CAR-T-cells bearing PD-1 chimeric cytokine receptor ectodomain variants following treatment with PD-L1-Fc ( FIG. 8A ) or anti-PD-1 ( FIG. 8B ). 2×PD-1 and 3×PD-1 chimeric cytokine receptor CAR-T-cells showed high levels of basal signaling, even in the absence of PD-L1 or anti-PD-1. Compared to WT PD-1 chimeric cytokine receptor CAR-T-cells, CAR-T-cells bearing the 2×PD-1 chimeric cytokine receptor, 3×PD-1 chimeric cytokine receptor and HA PD-1 chimeric cytokine receptor showed enhanced response to crosslinked PD-L1-Fc, with the HA PD-1 chimeric cytokine receptor providing the greatest response ( FIG. 8A ). In response to anti-PD-1 alone, the HA PD-1 chimeric cytokine receptor was the only ectodomain variant that induced increased pStat5; neither the 2×PD-1 chimeric cytokine receptor nor the 3×PD-1 chimeric cytokine receptor induced pStat5 beyond their levels of basal activity ( FIG. 8B ). Based on this analysis, the HA PD-1 chimeric cytokine receptor was the ectodomain variant that provided the lowest basal activity and greatest inducibility. 
     To evaluate the cytotoxic activity of PD-1 chimeric cytokine receptor CAR-T-cells, the U87KO-EGFRvIII-nucGFP target cell line that endogenously expresses PD-L1 and PD-L2. U87KO-EGFRvIII (gift from Cellectis SA (Paris, France)). U87KO-EGFRvIII was derived from the parental cell line, U87MG (ATCC), by first knocking out endogenous wild type EGFR using Transcription Activator-Like Effector Nucleases (TALEN), and then stably overexpressing full-length human EGFRvIII via lentiviral transduction. To facilitate target cell imaging via the IncuCyte Live Cell Analysis Imaging System, U87KO-EGFRvIII-nucGFP target cells were derived from U87KO-EGFRvIII by a second lentiviral transduction with IncuCyte NucLight Green Lentivirus Reagent (Sartorius). PD-L1 and PD-L2 expression on U87KO-EGFRvIII-nucGFP was determined by staining with an APC-conjugated anti-human PD-L1 (Biolegend) and a PE-conjugated anti-human PD-L2 (Biolegend), followed by flow cytometry analysis. For the in vitro cytotoxicity assay, 5,000 U87KO-EGFRvIII-nucGFP target cells were seeded and allowed to attach in 96-well plates with black walls and flat clear bottom in 50 uL RPMI containing 10% FBS (Hyclone), non-essential amino acids, sodium pyruvate and 20-25 mM HEPES. EGFRvIII CAR (2173 scFv) T-cells bearing the respective PD-1 chimeric cytokine receptor ectodomain variants were thawed and added to plated target cells at an Effector:Target (E:T) ratio of 1:4. FKBP chimeric cytokine receptor CAR-T-cells that do not activate cytokine signaling in response to PD-1 ligands were used as a control. Duplicate wells were set up for each condition. The number of live target cells at each timepoint was determined by enumerating the number of live nucGFP+target cells using the IncuCyte Live Cell Analysis Imaging System. 
       FIGS. 9A-B  show the cytotoxic response of the PD-1 chimeric cytokine receptor CAR-T-cells against target cells expressing PD-1 ligands.  FIG. 9A  shows that U87KO-EGFRvIII-nucGFP cells express high levels of endogenous PD-L1 and PD-L2.  FIG. 9B  shows the results of the in vitro cytotoxic assay against U87KO-EGFRvIII-nucGFP target cells. Compared to FKBP chimeric cytokine receptor CAR-T-cells, WT PD-1 chimeric cytokine receptor CAR-T-cells showed marginally improved cytotoxic responses. While the 2×PD-1 chimeric cytokine receptor did not enhance cytotoxicity beyond that of the WT PD-1 chimeric cytokine receptor, the HA PD-1 chimeric cytokine receptor significantly enhanced CAR-T-cell cytotoxicity. 
     Example 3: Optimization of the HA PD-1 Chimeric Cytokine Receptor 
     The HA PD-1 ectodomain variant afforded a significant enhancement over the WT PD-1 chimeric cytokine receptor. The helical transmembrane (TM) region of TpoR is critical for ligand-induced receptor signaling by controlling JAK2 activation. It was analyzed whether in a chimeric cytokine receptor where a different ectodomain (e.g. PD-1) is fused with the TpoR TM/JAK2-activating domain, the optimal structural conformation for receptor activation is perturbed. The responsiveness of the HA PD-1 chimeric cytokine receptor was attempted to be increased by modifying the TpoR TM region. To this end, HA PD-1 chimeric cytokine receptor variants with either deletions or insertions in the TpoR TM region were generated and tested in the HEK293T-cell reporter assay. 
     Briefly, 20,000 HEK293T-cells were plated into each well of a poly-L-lysine-coated 96-well flat-bottom plate and allowed to adhere overnight. A PD-1 chimeric cytokine receptor (2.5 ng), a Stat response element that drives Firefly Luciferase (100 ng; Promega) and Renilla Luciferase control reporter vector (1 ng; Promega) were mixed in a final volume of 5 uL in Opti-MEM (Gibco) (“DNA mix”). As a positive control, a FKBP chimeric cytokine receptor was usedwe, in which the WT PD-1 ectodomain was replaced with FKBP12(F36V), so that cytokine signaling could be inducible by the small molecule, AP1903. 0.3 uL Lipofectamine 2000 (Invitrogen) in 5 uL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 10 uL was added to each well containing HEK-293T. 24 hours after transfection, cells were either left untreated, or treated with the indicated concentrations of anti-PD-1 (Nivolumab; Selleck Chemical), recombinant human PD-L1-Fc (Biolegend), anti-Human IgG Fc gamma secondary antibody (Thermo Fisher), or AP1903 (Apex Bio). As negative controls, hIgG4 and hIgG1 isotype controls were added in place of anti-PD-1 and PD-L1-Fc, respectively. All treatments were diluted in serum-free media. 24 hours after treatment, Stat reporter activity was evaluated using the Dual-Glo Luciferase Assay System (Promega). Fold induction of Stat5 reporter activity was normalized to that of HEK293T-cells transfected with all vectors except for the chimeric cytokine receptor receptor and that were left untreated. 
       FIGS. 10A-B  show the amino acid sequences for the wild type TpoR and the various TM deletion ( FIG. 10A ) or insertion ( FIG. 10B ) variants. 
       FIGS. 11A-B  show the response of a HA PD-1 chimeric cytokine receptor bearing TpoR TM deletions ( FIG. 11A ) and extensions ( FIG. 11B ) following treatment with PD-L1-Fc. Unlike the HA PD-1 chimeric cytokine receptor that did not respond to PD-L1-Fc alone, the N-6, N-7, N-8, N-9 and N+4 TpoR TM variants were able to induce Stat5 activity in response to PD-L1 alone. Furthermore, compared to the HA PD-1 chimeric cytokine receptor, the N-5, N-6, N-7, N-8, N-9 and N+4 TpoR TM variants improved Stat5 activity in response to high concentrations of crosslinked PD-L1. The N-9 and N+4 TpoR TM variants retained the ability to respond to low concentrations of crosslinked PD-L1. These demonstrate that modulating the TpoR TM region enhanced the responsiveness and sensitivity of the HA PD-1 chimeric cytokine receptor to PD-1 ligands. 
       FIGS. 12A-B  shows the response of HA PD-1 chimeric cytokine receptor bearing TpoR TM deletions ( FIG. 12A ) and extensions ( FIG. 12B ) following treatment with anti-PD-1 (Nivolumab). Unlike the HA PD-1 chimeric cytokine receptor that did not respond to anti-PD-1 alone, the N-5, N-6, N-7, N-8, N-9 and N+4 TpoR TM variants were able to induce Stat5 activity in response to anti-PD-1 alone, without the need for further crosslinking. These demonstrate that modulating the TpoR TM region enhanced the responsiveness and sensitivity of the HA PD-1 chimeric cytokine receptor to anti-PD-1. 
     Example 4: Effect of the HA PD-1 Chimeric Cytokine Receptor on CAR-T-Cell Cytotoxic Activity 
     PD-1 blockade strategies, such as combination with anti-PD-1 or forced overexpression of a dominant-negative (DN) PD-1 decoy receptor, can ameliorate CAR-T-cell exhaustion. While PD-1 blockade prevents a negative signal from being transmitted into CAR-T-cells, it does not actively transmit any positive signals. It was therefore tested whether the HA PD-1 chimeric cytokine receptor, which couples PD-1 inhibition with simultaneous immune-potentiating cytokine signals, is more effective than PD-1 blockade alone. The cytotoxic activity of HA PD-1 chimeric cytokine receptor CAR-T-cells was compared against CAR-T-cells combined with Nivolumab, and against CAR-T-cells expressing a dominant-negative (DN) PD-1. (amino acid sequences in Table 9). 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 PD1 dominant negative (DN) sequences: 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ ID 
               
               
                 PD1 DN 
                 Amino acid sequence 
                 NO: 
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 129 
               
               
                 PD1(21-207) 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 (Dominant 
                 VLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGR 
                   
               
               
                 negative wild 
                 DFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTE 
                   
               
               
                 type PD1) 
                 RRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWV 
                   
               
               
                   
                 LAVICSRAARGTI GARRTGQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 130 
               
               
                 HAPD1 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 (Dominant 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 negative high 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 affinity PD1) 
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                   
                 AVICSRAARGTI GARRTGQ 
               
               
                   
               
            
           
         
       
     
     To assess in vitro cytotoxicity, 5,000 PD-L1+/PD-L2+U87KO-EGFRvIII-nucGFP target cells were seeded and allowed to attach in 96-well plates with black walls and flat clear bottom in 50 uL RPMI containing 10% FBS (Hyclone), non-essential amino acids, sodium pyruvate and 20-25 mM HEPES. EGFRvIII CAR (2173 scFv) T-cells bearing the respective PD-1 chimeric cytokine receptors or DN PD-1 were thawed and added to plated target cells at an Effector:Target (E:T) ratio of 1:4. FKBP chimeric cytokine receptor CAR-T-cells that do not activate cytokine signaling in response to PD-1 ligands were used as a control. Where indicated, anti-PD-1 (Nivolumab) or hIgG4 isotype control antibodies were added at a concentration of 10 ug/mL. Duplicate wells were set up for each condition. The number of live target cells at each timepoint was determined by enumerating the number of live nucGFP+target cells using the IncuCyte Live Cell Analysis Imaging System. 
       FIGS. 13A-B  show the cytotoxic response of PD-1 chimeric cytokine receptor CAR-T-cells in the absence ( FIG. 13A ) or presence ( FIG. 13B ) of anti-PD-1. Although combination with anti-PD-1 did improve the activity of FKBP chimeric cytokine receptor CAR-T-cells and WT PD-1 CAR-T-cells, HA PD-1 chimeric cytokine receptor CAR-T-cells were more effective at target cell lysis. Combination with anti-PD-1 did not further improve the activity of HA PD-1 chimeric cytokine receptor CAR-T-cells, indicating that PD-1 ligands expressed on the target cells could bind to and activate the PD-1 chimeric cytokine receptor. 
       FIG. 14  shows the cytotoxic response of CAR-T-cells coexpressing either a DN WT PD-1, a DN HA PD-1 or the HA PD-1 chimeric cytokine receptor. HA PD-1 DN CAR-T-cells were more effective than WT PD-1 CAR-T-cells. Without being bound by any theory, this could be due to more effective sequestration and blockade of PD-1 ligands. However, HA PD-1 chimeric cytokine receptor CAR-T-cells were more cytotoxic than HA PD-1 DN CAR-T-cells. Without being bound by any theory, this could be due to the coupling of PD-1 blockade with simultaneous cytokine signaling. 
     Example 5: PD-1 Chimeric Cytokine Receptor that Delivers Constitutive Cytokine Signals in a Ligand-Independent Fashion 
     The TM/JAK2-activating domain of PD1 chimeric cytokine receptors in some of the examples above is derived from TpoR, whose signaling activity can be modulated by key residues within the TpoR transmembrane (TM) domain. For instance, single point mutations within the TpoR TM domain, such as at S505 or W515, have been reported to cause constitutive TpoR signaling; on the other hand, mutating H499 in the TpoR TM domain dampens signaling from these activating mutations (Proc Natl Acad Sci USA. 2013 Feb. 12; 110(7):2540-5.; FASEB J. 2011 July; 25(7):2234-44.; J Biol Chem. 2016 Feb. 5; 291(6):2974-87.). It was analyzed whether a PD-1 chimeric cytokine receptor can be constitutively active by introducing the above-described single point mutations into the TpoR TM region of the receptor. To this end, HA PD-1 chimeric cytokine receptor variants with constitutively active single point mutations (S505 and/or W515) of the TpoR TM region and a single point mutation that reduces the activity of the TpoR TM region were generated. To mimic signaling from either IL-7 or IL-2/IL15, recruiting domains derived from the IL-7Ra and IL-2/IL15Rb receptors, respectively, were fused downstream of the TpoR TM/JAK2-activating domain. Exemplary amino acid sequences of the constructs are in Table 10. 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 PD1 chimeric cytokine receptor sequences: 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ ID 
               
               
                 Receptor 
                 Amino acid sequence 
                 NO: 
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 141 
               
               
                 HAPD1.TPOR/ 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 MPLR(478- 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 582; H499L, 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 S505N, 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 W515K).IL7Ra 
                 SDPTRVETATETAWISLVTAL   L   LVLGL   N   AVLGLLLLR   K   QFPA 
                   
               
               
                 (316-459) 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 142 
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 (478-582; 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 S505N, W515K). 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 IL2Rbsmall 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 (393-433, 
                 SDPTRVETATETAWISLVTALHLVLGLNAVLGLLLLRKQFPA 
                   
               
               
                 518-551) 
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 143 
               
               
                 HAPD1DN 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                   
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                   
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                   
                 RAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVL 
                   
               
               
                   
                 AVICSRAARGTI GARRTGQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 144 
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 (478- 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 582).IL7Ra 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (316-459) 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                   
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 ARDEVEGFLQDTFPQQLEESEKQRLGGDVQSPNCPSEDVVIT 
                   
               
               
                   
                 PESFGRDSSLTCLAGNVSACDAPILSSSRSLDCRESGKNGPHV 
                   
               
               
                   
                 YQDLLLSLGTTNSTLPPPFSLQSGILTLNPVAQGQPILTSLGSN 
                   
               
               
                   
                 QEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARP 
                 145 
               
               
                 HAPD1.TpoR 
                 PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESF 
                   
               
               
                 (478-582). 
                 HVIWHRESPSGQTDTLAAFPEDRSQPGQDCRFRVTQLPNGRD 
                   
               
               
                 IL2Rbsmall 
                 FHMSVVRARRNDSGTYVCGVISLAPKIQIKESLRAELRVTER 
                   
               
               
                 (393-433, 
                 RAEVPTAHPSPSPRPAGQFQTLV 
                   
               
               
                 518-551) 
                 SDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQFPA 
                   
               
               
                   
                 HYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSDTC 
                   
               
               
                   
                 EEVEPSLLEILPKSSERTPLPL 
                   
               
               
                   
                 DEGVAGAPTGSSPQPLQPLSGEDDAYCTFPSRDDLLLFSPSG 
                   
               
               
                   
                 QGEFRALNARLPLNTDAYLSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARPPGWFLDSPDRPWNPPTFSPAL 
                 210 
               
               
                 HAPD1.TPOR/ 
                 LVVTEGDNATFTCSFSNTSESFHVIWHRESPSGQTDTLAAFPE 
                   
               
               
                 MPLR(478- 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYVCG 
                   
               
               
                 582; H499L, 
                 VISLAPKIQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQT 
                   
               
               
                 S505N, 
                 LVSDPTRVETATETAWISLVTALLLVLGLNAVLGLLLLRKQF 
                   
               
               
                 W515K).IL7Ra 
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                 (316-459) 
                 TCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFLQDTFPQQLE 
                   
               
               
                   
                 ESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLAGNVS 
                   
               
               
                   
                 ACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNSTLPPP 
                   
               
               
                   
                 FSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARPPGWFLDSPDRPWNPPTFSPAL 
                 211 
               
               
                 HAPD1.TpoR 
                 LVVTEGDNATFTCSFSNTSESFHVIWHRESPSGQTDTLAAFPE 
                   
               
               
                 (478-582; 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYVCG 
                   
               
               
                 S505N, W515K). 
                 VISLAPKIQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQT 
                   
               
               
                 IL2Rbsmall 
                 LVSDPTRVETATETAWISLVTALHLVLGLNAVLGLLLLRKQF 
                   
               
               
                 (393-433, 
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                 518-551) 
                 TCEEVEPSLLEILPKSSERTPLPL LE DEGVAGAPTGSSPQPLQP 
                   
               
               
                   
                 LSGEDDAYCTFPSRDDLLLFSPSGQGEFRALNARLPLNTDAY 
                   
               
               
                   
                 LSLQELQGQDPTHLV 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARPPGWFLDSPDRPWNPPTFSPAL 
                 212 
               
               
                 HAPD1.TpoR 
                 LVVTEGDNATFTCSFSNTSESFHVIWHRESPSGQTDTLAAFPE 
                   
               
               
                 (478- 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYVCG 
                   
               
               
                 582).IL7Ra 
                 VISLAPKIQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQT 
                   
               
               
                 316-459) 
                 LVSDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQF 
                   
               
               
                   
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                   
                 TCEEVEPSLLEILPKSSERTPLPL LE ARDEVEGFLQDTFPQQLE 
                   
               
               
                   
                 ESEKQRLGGDVQSPNCPSEDVVITPESFGRDSSLTCLAGNVS 
                   
               
               
                   
                 ACDAPILSSSRSLDCRESGKNGPHVYQDLLLSLGTTNSTLPPP 
                   
               
               
                   
                 FSLQSGILTLNPVAQGQPILTSLGSNQEEAYVTMSSFYQNQ 
                   
               
               
                   
               
               
                 CD8SS- 
                 MALPVTALLLPLALLLHAARPPGWFLDSPDRPWNPPTFSPAL 
                 213 
               
               
                 HAPD1.TpoR 
                 LVVTEGDNATFTCSFSNTSESFHVIWHRESPSGQTDTLAAFPE 
                   
               
               
                 (478-582). 
                 DRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYVCG 
                   
               
               
                 IL2Rbsmall 
                 VISLAPKIQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQT 
                   
               
               
                 (393-433, 
                 LVSDPTRVETATETAWISLVTALHLVLGLSAVLGLLLLRWQF 
                   
               
               
                 518-551) 
                 PAHYRRLRHALWPSLPDLHRVLGQYLRDTAALSPPKATVSD 
                   
               
               
                   
                 TCEEVEPSLLEILPKSSERTPLPL LE DEGVAGAPTGSSPQPLQP 
                   
               
               
                   
                 LSGEDDAYCTFPSRDDLLLFSPSGQGEFRALNARLPLNTDAY 
                   
               
               
                   
                 LSLQELQGQDPTHLV 
               
               
                   
               
            
           
         
       
     
     A HEK293T cell reporter assay was used to test for constitutive cytokine signaling by PD1 chimeric cytokine receptors. Briefly, 20,000 HEK293T cells were plated into each well of a poly-L-lysine-coated 96-well flat-bottom plate and allowed to adhere overnight. A PD1 chimeric cytokine receptor-CAR construct (2.5 ng), a Stat5 response element that drives Firefly Luciferase (100 ng; Promega) and Renilla Luciferase control reporter vector (1 ng; Promega) were mixed in a final volume of 5 μL in Opti-MEM (Gibco) (“DNA mix”). As negative controls, cells were transfected with either a BFP CAR construct that expresses BFP instead of a PD1 chimeric cytokine receptor, or a dominant negative HA PD1 ectodomain CAR construct. As a comparison, cells were transfected with a vector encoding an inducible PD1 chimeric cytokine receptor (SEQ ID NOs: 213-214). As a positive control, cells were transfected with a vector encoding full-length human EpoR (in place of the cytokine receptor-CAR construct) so that Stat5 signaling could be induced by the addition of exogeneous recombinant human Epo. 0.3 μL Lipofectamine 2000 (Invitrogen) in 5 μL Opti-MEM was incubated at room temperature for 5 minutes and then added to the DNA mix. The mixture was incubated at room temperature for 20 minutes and the total volume of 10 μL was added to each well containing HEK-293T. 24 hours after transfection, human PD-L1 Fc, a secondary crosslinking antibody, or an isotype control antibody were added at the indicated final concentrations. 48 hours after transfection, Stat5 reporter activity was evaluated using the Dual-Glo Luciferase Assay System (Promega). Fold induction of Stat5 reporter activity was normalized to that of HEK293T cells transfected with all vectors except for the cytokine receptor and that were left untreated. 
       FIG. 15  shows Stat5 reporter activity in the absence and presence of PD-L1. As expected, negative controls (i.e. BFP CAR and dominant negative HA PD1 ectodomain CAR constructs) did not induce Stat5 reporter activity. The inducible PD1 chimeric cytokine receptors bearing a TM/JAK2-activating domain derived from wild type TpoR ((SEQ ID NOs: 213-214) was active in the presence of crosslinked PD-L1, but was inactive in the absence of ligand. In contrast, strong, constitutive signaling was generated from PD1 chimeric cytokine receptors comprising a TM/JAK2-activating domain derived from a TpoR transmembrane domain with constitutively active point mutations (SEQ ID NOs.: 210-211) in a ligand-independent fashion. This ligand-independence is in contrast to an inducible PD1 chimeric cytokine receptor—bearing the wild type TpoR transmembrane domain—where the presence of PD1 ligands is required for cytokine signaling to occur. The constitutive activity of the PD1 chimeric cytokine receptor comprising the TM/JAK2-activating domain derived from a TpoR transmembrane domain with constitutively active point mutations (SEQ ID NO.: 2211) was increased in the presence of PD-L1 and crosslinked PD-L1. The activity of the PD1 chimeric cytokine receptor that comprises the constitutively active TpoR TM mutations and the inhibitor mutation (H499) (SEQ ID NO.: 210) showed a slight dampening of the constitutive activity in the presence of PD-L1 and crosslinked PD-L1. 
     Example 6 Increasing the Sensitivity and Responsiveness of the HA PD1 Switch Further Improves CAR T Cell Cytotoxicity 
     As demonstrated above, the HA PD1 switch could be engineered for increased sensitivity and responsiveness towards PD-1 ligands and anti-PD-1 by modifying the TpoR TM helix. Specifically, in the HEK293T cell reporter assay, the N-5, N-6, N-7, N-8, N-9 and N+4 TpoR TM variants enhanced STAT5 activation in response to crosslinked PD-L1 ( FIG. 11 ) and anti-PD-1 ( FIG. 12 ). To evaluate if this translated into improved CAR T cell cytotoxicity in the presence of PD-1 ligands and anti-PD-1, we compared the killing activity of HA PD1 switch CART cells bearing either the wildtype (WT) or variant TpoR TM domains. 
     Briefly, 5,000 PD-L1+/PD-L2+U87KO-EGFRvIII-nucGFP target cells were seeded and allowed to attach in 96-well plates with black walls and flat clear bottom in 50 uL RPMI containing 10% FBS (Hyclone), non-essential amino acids, sodium pyruvate and 20-25 mM HEPES. EGFRvIII CAR (2173 scFv) T cells coexpressing a HA PD1 switch with the indicated TpoR TM domain were thawed and added to plated target cells at an Effector:Target (E:T) ratio of 1:4 or 1:8. As a control, CAR T cells coexpressing BFP in place of a HA PD1 switch, and that do not activate cytokine signaling in response to PD-1 ligands, were used. Where indicated, anti-PD-1 (nivolumab) or hIgG4 isotype control antibodies were added at a concentration of 10 ug/mL. Duplicate wells were set up for each condition. The number of live target cells at each timepoint was determined by enumerating the number of live nucGFP+target cells using the IncuCyte Live Cell Analysis Imaging System. 
       FIG. 16  shows the cytotoxic activity of HA PD1 switch CART cells with the indicated TpoR TM domains at an E:T ratio of 1:4 and in the absence of anti-PD-1. As expected, compared to the control BFP-CAR T cells, HA PD1 switch CAR T cells with the WT TpoR TM domain showed improved target cell lysis in vitro. However, cytotoxic activity was further enhanced in HA PD1 switch CAR T cell variants bearing the N-5, N-7, N-8, N-9 or N+4 TpoR TM domains. Concordant with  FIGS. 11 and 12 , these data indicate that TpoR TM domain variants allow more effective cytotail activation in response to PD-1 ligands, thereby further enhancing the cytotoxic potency of HA PD1 switch CAR T cells. 
       FIGS. 17A-B  show the cytotoxic activity of HA PD1 switch CAR T cells with the indicated TpoR TM domains at an E:T ratio of 1:8, and in the absence or presence of anti-PD-1.  FIG. 17A  shows HA PD1 switch CART cell cytotoxicity in the absence of anti-PD-1. As expected, target cell lysis was less efficient at this lower E:T ratio as compared to the 1:4 ratio. Compared to the control BFP-CAR T cells, co-expression of a HA PD1 switch improved CAR T cell cytotoxicity, with the TpoR TM variants showing better activity than their WT TpoR TM counterpart.  FIG. 17B  shows HA PD1 switch CART cell cytotoxicity in the presence of anti-PD-1. Cytotoxic activity of control BFP-CAR T cells was not markedly improved even when combined with anti-PD-1. Additional dimerization and activation by anti-PD-1 improved the activity of HA PD1 switch with WT TpoR TM domain, and this was further enhanced in the N-7, N-8 and N-9 TpoR TM variants.  FIGS. 17C-E  show a comparison of the N-7 ( FIG. 17C ), N-8 ( FIG. 17D ) and N-9 ( FIG. 17E ) TpoR TM variants in the absence or presence of anti-PD-1. Without being limited to any specific mechanisms, the engagement of the PD-1 ectodomain with monomeric PD-L1/2 expressed on tumor cells or with a dimeric anti-PD-1 antibody may affect the conformation of the TpoR TM helix and different strength of signal transduction. Taken together, increasing the sensitivity and responsiveness of the HA PD1 switch via TpoR TM modifications can markedly improve the efficiency of cytotail signaling and enhance CAR T cell cytotoxic potency. 
     All references cited herein, including patents, patent applications, papers, text books, and the like, and the references cited therein, to the extent that they are not already, are hereby incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls. 
     The foregoing description and Examples detail certain specific embodiments of the invention and describes the best mode contemplated by the inventors. It will be appreciated, however, that no matter how detailed the foregoing may appear in text, the invention may be practiced in many ways and the invention should be construed in accordance with the appended claims and any equivalents thereof.