bluuebunny/crossref_metadata_2025_split huggingface dataset:
CCR2 identifies first responders among human CD4+ memory T cells: long-lived, apoptosis-resistant, antigen-responsive cells with enhanced migration potential, a low threshold for activation, and immediate effector capabilities (85.5)
Authors: Farber Joshua M. , Song Kaimei , Zhang Hongwei H. , Rabin Ronald L. , Hill Brenna J. , Sereti Irini , Prussin Calman , Siegel Richard M. , Douek Daniel C. , Roederer Mario 
Published: January 2023

Abstract
The persistence and roles of memory T cells with effector capability have been questioned. We analyzed human memory CD4+ T cells from peripheral blood defined by their expression of chemokine receptors CCR5 and CCR2, the principal subsets being CCR5−CCR2−, CCR5+CCR2−, and CCR5+CCR2+. CCR2+ cells co-expressed the most chemokine receptor types (up to six) and migrated to the greatest number of chemokines, followed by CCR5+CCR2− cells. Numbers of T cell receptor gene excision circles were CCR5−CCR2− &gt; CCR5+CCR2− &gt; CCR5+CCR2+. The CCR2+ cells were those most readily activated through TCR but showed reduced proliferative potential, were enriched in cells responding to a remote immunogen, secreted high levels of effector cytokines, and were resistant to apoptosis. By contrast, the CCR5+CCR2− population was enriched in recently activated/cycling cells and Treg, and was more susceptible to apoptosis. The data suggest that patterns of CCR5 and CCR2 expression separate effector- vs. memory-cell enriched CD4+ subsets, and that CCR2 marks highly differentiated, long-lived memory cells with effector capabilities. The data also suggest co-ordination among a memory cell’s position on a unidirectional pathway of differentiation, ability to be recruited into tissue, and threshold for activation/effector function, with the most chemokine-responsive cells being best equipped as first responders in a recall response.
Identification of a mouse mutant with highly deficient cytotoxic T cell memory generation using ENU-induced genome-wide mutagenesis (85.7)
Authors: Kung John T. 
Published: January 2023

Abstract
While much has been learned about CTL memory maintenance, much remains to be learned on the early events of CTL activation or induction phase. Through an ENU-induced genome-wide mouse mutagenesis program, we screened G3 mice using CD44 expression on CD8+ T cells as a surrogate indicator of the relative representation of the CTL memory subset. One such identified mutant (P-054) has highly deficient CD44hi subset within CD8+ T cells but normal CD44hi subset in CD4+ T cells. An affected male mouse was bred to normal females to generate F1 mice that all had normal numbers of CTL memory subset based on CD44 expression. Random intercross of F1 mice generated CTL deficienct F2 mice at a ratio of ~25%, indicating heritability of this trait and recessive transmission. Linkage of the mutant gene has been established to chromosome 4 to within a region of ~30 genes. The mutant mouse (H-2b) was immunized by BALB.B spleen cells (H-2b) and CTL memory T cells were identified by H60 (a BALB/c-encoded minor histocompatibility ag)-sepcific CTL by appropriate pentamers and a highly deficient H60-specific CTL response was seen. Adoptive transfer of 2C TCR tg CD8+ T cells activated in the presence of added IL-4, a condition known to promote strong CTL memory generation, into P-054 mice resulted in potent maintenance of 2C memory T cells. Thus, P-054 can provide an environment required to maintain known memory CD8+ T cells and likely represents a novel mutant model that is deficient in the induction phase of CTL memory generation.
B cell receptor signaling governs the subcellular location of Toll-like receptor 9 (86.11)
Authors: Chaturvedi Akanksha , Pierce Susan 
Published: January 2023

Abstract
Toll-like receptor 9 (TLR9), an innate immune system receptor for hypomethylated CpG DNA motifs, has gained substantial attention recently due to its unique ability to promote the activation of autoreactive B cells in response to DNA-containing antigens. Many of the autoantigens targeted in systemic lupus erythematosus (SLE) contain DNA. Moreover, TLR9 has been shown to play a critical role in regulating anti-DNA autoantibody production in mouse models of lupus by mechanisms that involve the simultaneous engagement of TLR9 and the B cell receptor (BCR). However, the cellular and molecular basis of the interaction between TLR9 and the BCR is not known and given the spatial segregation of the BCR on the plasma membrane and TLR9 in early endosomal compartments, it is not obvious how synergy is achieved. Here we show that when activated independently the BCR and TLR9 signal from discrete subcellular compartments, the BCR initially from the plasma membrane and later from large multivesicular bodies (MVB) and TLR9 from multiple endosomes. Remarkably, stimulation of B cells simultaneously through the BCR and TLR9 triggers the recruitment of the TLR9-containing endosomes to the MVB by a Src-family kinase-dependent, MyD88-independent mechanism resulting in a synergistic response. Thus, the BCR triggers the recruitment of TLR9- containing vesicles to the MVB presumably to allow TLR9 to survey the BCR bound antigen for its DNA ligand, which if present, results in TLR9 activation and enhanced B cell responses. Our study adds a new spatial dimension to our view of TLR9 signaling in the B cells, potentially providing a new arena for therapeutic drug design.
CS1 (CRACC) induces Proliferation and Autocrine Cytokine expression on Activated human B Lymphocytes (86.3)
Authors: LEE JAE KYUNG , Vaidya Swapnil V , Kumaresan Pappanaicken R , Mathew Porunelloor A 
Published: January 2023

Abstract
CS1 (CRACC, CD319), a member of the CD2 family of cell surface receptors is implicated in the activation of NK cell- mediated cytotoxicity. Previous studies showed that CS1 is also expressed on activated B cells. However, the functional role of CS1 in human B-lymphocytes is not known. Two isoforms of CS1, CS1-L and CS1-S are expressed in human NK cells that differentially regulate NK cell function. CS1-L contains immunoreceptor tyrosine-based switch motifs (ITSMs) in its cytoplasmic domain whereas CS1-S lacks ITSM. Here we show that human B lymphocytes express only CS1-L isoform and its expression is upregulated upon B cell activation with various stimulators. Moreover, anti-CS1 mAb strongly enhanced proliferation of both freshly isolated as well as activated B cells. The enhanced proliferation effects of CS1 were most prominent on B cells activated by anti-CD40 mAbs and/or hrIL-4. Human cytokine microarray and quantitative real-time PCR results indicated that CS1 activation enhanced mRNA transcripts of flt3 ligand, lymphotoxin A, TNF, and IL-14. These results suggest that activation of B lymphocytes through surface CS1 may be mediated through secretion of autocrine cytokines and CS1 may play a role in the regulation of B lymphocytes proliferation during immune responses.
Intracellular signaling pathways triggered by activation of the Epstein-Barr virus receptor (gp140, CR2, CD21, EBV/C3dR) on cell surface of human B lymphocytes (86.5)
Authors: FRADE Raymond , LOTTIN Severine 
Published: January 2023

Abstract
We originally isolated gp140, the receptor of human C3 (the third component of complement) of human B lymphocytes (1981). gp140 is the C3d receptor (1985a) and also the Epstein-Barr virus receptor (1985b). EBV/C3dR regulates proliferation (1985c) and transformation (1985b) of human B lymphocytes. Thus, we analyzed intracellular events triggered by activated EBV/C3dR. EBV/C3dR activation triggered its intracellular interactions with a p120 nuclear RNP (1987) and the p53 oncoproteine (1995). Then, we first identified two signaling pathways specifically triggered by activated EBV/C3dR. The first pathway is characterized by PI-3 kinase activation:
upstream PI-3 kinase, pp60src kinase is activated to tyrosine phosphorylate nucleolin. Then, tyrosine phosphorylated nucleolin interacts with SH2 domains of the p85 subunit of PI-3 kinase, activating this latter;downstream PI 3-kinase, AKT-GSK3 pathway is activated. This pathway is independent of BCR and CD19 activations (1999, 2001, 2003).
The second pathway is characterized by:
Cbl phosphorylation;Cbl interactions with Crk-L, Syk and the p85 subunit of PI-3-kinase;dissociation of the Cbl-Vav complex (2006).
This dissociation, in opposite to Cbl-Vav association triggered by BCR and CD19, emphasizes the specificity of EBV/C3dR signaling. We herein identified the functional relationship between both pathways.
This work is supported by INSERM
T Cell Receptor-induced NF-KB Activation Is Negatively Regulated by E3 Ubiquitin Ligase Cbl-b via Akt- and PKC-theta-dependent Pathways (87.10)
Authors: Qiao Guilin , Li zhenping , Alegre Maria-Luisa , Ying Haiyan , Zhang Jian 
Published: January 2023

Abstract
Previously, we demonstrated that Cbl-b tunes the threshold for T cell activation mediated by CD28 and CTLA-4. Cbl-b negatively regulates Vav-1 via a PI3-K-dependent mechanism. However, it is unknown whether Cbl-b also targets other signaling pathway(s). In this study, we found that loss of Cbl-b selectively results in aberrant activation of TCR-induced NF-κB which is mediated by PI3-K/Akt and PKC-𝛉. The absence of Cbl-b also results in enhanced TCR-induced activation of PLC-γ1 which may selectively induce activation of PKCs including PKC-𝛉 but not Ca2+ signaling pathway. Cbl-b associates with PKC-𝛉 upon TCR stimulation and regulates TCR-induced PKC-𝛉 activation through Vav-1 which couples PKC-𝛉 to PI3-K and allows it to be phosphorylated possibly by PDK-1. PKC-𝛉 then couples IKKs to CBM complex, resulting in activation of IKK complex. Therefore, our data first demonstrate that down-regulation of TCR-induced NF-κB activation by Cbl-b is coordinately mediated by both Akt- and PKC-𝛉dependent signaling pathways in primary T cells.
Measurement of nuclear translocation in primary cells using correlation analysis of images obtained on the ImageStream imaging flow cytometer (87.13)
Authors: George Thaddeus , Harmon Ian , Peterson Erik J , Burbach Brandon J , Shimizu Yoji , Matsuda Jennifer L , Gapin Laurent , Connors Mark , Dowdell Kennichi C , Farrar J David , Hall Brian E , Morrissey Philip J 
Published: January 2023

Abstract
Signal transduction pathways regulate T cell activation and differentiation and have a critical influence on the outcome of adaptive immune responses. It has recently been shown that nuclear translocation events within non-adherent tumor cells can be quantified by correlating transnscription factor and nuclear images collected on the ImageStream multispectral imaging flow cytometer. Here we extend the quantitative technique to the study of nuclear localization within primary human or murine T cells in the following systems:
translocation of T-bet in CD4 cells derived from murine spleen and lymph node cellstranslocation of NFAT in tetramer positive CD8 cells from human peripheral blood,translocation of NFκB in CD4+CD8+ thymocytes involved in conjugates with APC presenting cognate peptide, andlocalization of phosphorylated STAT4 in human CD4 cells following cytokine stimulation.
These data demonstrate image-based nuclear localzation within primary cells with large nuclear to cytoplasmic area ratios using ImageStream technology.
Ets1-dependent IP3R3 expression in naïve CD4+ T cells is required for cytokine gene expression (87.22)
Authors: NAGALEEKAR VISWAS K , DIEHL SEAN , Juncadella Ignacio , Charland Colette , Garrett-Sinha Lee Ann , Muthusamy Natarajan , Anguita Juan , Rincón Mercedes 
Published: January 2023

Abstract
IP3 receptors (IP3Rs) are critical for the release of Ca++ from intracellular stores in response to IP3 generated upon T cell receptor (TCR) ligation. However, little is known about the expression of the different IP3Rs in CD4+ T cells and their contribution to cytokine gene expression during antigen stimulation. Here, we show for the first time that prior to activation, naïve CD4+ T cells only express IP3R3, but not IP3R1 and IP3R2. IP3R3-mediated Ca++ flux for cytokine gene expression is required for an extended period of time on the order of hours that varies for specific cytokine. IP3R3 gene expression in CD4+ T cells is dependent on the transcription factor Ets1 and is downregulated during the activation due to the loss of this transcription factor. The downregulation of IP3R3 in activated cells correlates with the failure of TCR ligation to trigger intracellular Ca++ mobilization in these cells. Thus, IP3R3 plays an important role in cytokine gene expression during early activation of naïve CD4+ T cells.
Endogenously produced IL-4 strongly enhances CD8+ T cell proliferation and accumulation in homeostatic conditions and during Th2 responses (87.24)
Authors: Morris Suzanne C. , Heidorn Stephanie M. , Yang Jun-Qi , Herbert De’Broski R. , Khodoun Marat V. , Finkelman Fred D. 
Published: January 2023

Abstract
IL-7 and IL-15 have important stimulatory roles in CD8+ T cell homeostasis. We evaluated whether another γc-related cytokine, IL-4, also stimulates CD8+ T cell proliferation and accumulation in vivo. Wild-type (WT) mice have twice as many splenic CD8+ T cells as IL-4- or IL-4Rα-deficient mice, with similar numbers of naïve cells but ~5-fold more memory phenotype cells. WT CD8+ T cells outcompete IL-4Rα-deficient CD8+ T cells following cell transfer and SCID mice reconstituted with equal numbers of WT and IL-4Rα-deficient CD8+ T cells have twice as many splenic WT as IL-4Rα-deficient CD8+ T cells 2 weeks later. WT, but not IL-4Rα-deficient CD8+ T cells proliferate when transferred into IL-4Rα-deficient mice that have been immunized with goat anti-mouse IgD antibody or infected with Nippostrongylus brasiliensis to stimulate a Th2 response. Consistent with this, splenic CD8+ T cell accumulation is entirely IL-4- and IL-4Rα-dependent and proliferation is predominantly IL-4- and IL-4Rα-dependent in mice infected w132th Schistosoma mansoni and pulmonary CD8+ T cell proliferation is rapidly suppressed by anti-IL-4R□ mAb in a mouse model of asthma. Furthermore, even ng quantities of IL-4 stimulate CD8+ T cell proliferation and accumulation in vivo. Thus, IL-4 is a potent, biologically important stimulator of CD8+ T cell proliferation and accumulation in vivo.
Supported by the US Dept of Veterans Affairs and NIH.
Enhanced development of memory like CD8 T cells with innate immune function in the absence of Tec kinase signals (87.26)
Authors: Hu Jianfang , August Avery 
Published: January 2023

Abstract
T cells with a memory phenotype and innate immune function have been reported, but the signals regulating these cells are still unclear. The Tec family kinases are highly expressed in T cells and involved in T cell activation and differentiation. We show here that Itk null mice have a consistent increase in the proportions and absolute numbers of CD8+ T cells that have memory phenotype (CD8+ CD44highCD122high). This elevation is observed in mice carrying an Itk mutant lacking the kinase domain, but not in mice carrying a transgene of the related kinase Rlk/Txk, which indicates that active Tec kinase signaling suppresses their presence. These cells carry preformed message for IFN-gamma and are able to produce IFN-gamma directly ex vivo. They also carry abundant transcripts for Granzyme B and eomesodermin. We also show that memory phenotype CD8+ T cells rapidly secrete IFN-gamma after infection with Listeria monocytogenes (LM) and Itk null mice have the ability to effectively response to infection. Antibody-mediated depletion of CD8+ T cells resulted in significantly reduced ability to clear LM infection in both WT and Itk null mice. Furthermore, transfer of small number of memory phenotype CD8+ T cells can reduce bacterial counts in IFN-gamma deficient mice 3 days after LM infection. These results indicate that these cells are functional CD8+CD44+ cells with innate like function and that the active signals from Tec kinases regulate their development.
Stomatin-Like Protein 2 Sustains Signalling from T Cell Receptor Signalosomes (87.36)
Authors: Kirchhof Mark G , Chau Luan A , Lemke Caitlin D , Vardhana Santosh , Darlington Peter J , Márquez Maria E. , Taylor Roy , Rizkalla Kamilia , Blanca Isaac , Dustin Michael L , Madrenas Joaquín 
Published: January 2023

Abstract
T cell activation involves the formation of an organized interface between the T cell and the antigen-presenting cell known as the immunological synapse (IS). Sustained T cell signalling at the IS emanates from peripheral supramolecular activating complexes (pSMACs). We hypothesized that a molecular machinery would anchor the TCR microclusters in the periphery of the synapse to the cytoskeleton allowing the recruitment of signalosome components and providing sustained TCR signalling. Here, we report that stomatin-like protein-2 (SLP-2) is a key element of this machinery by linking the components of the pSMACs to the actin cytoskeleton. SLP-2 is expressed at low levels in resting human lymphocytes but is rapidly up-regulated upon activation. In T cells, SLP-2 polarizes to the IS and actively accumulates in the pSMACs, within lipid rafts. SLP-2 sequentially associates with CD3-ε, Lck, ZAP-70, phospho-LAT and phospho-PLC-γ1, as well as with polymerized actin. In vivo, SLP-2 expression can be detected in areas of lymphocyte activation in human lymph nodes and thymi. Consistent with these data, up-regulation of SLP-2 in T cells correlates with enhanced activation, whereas knocking-down SLP-2 expression in human effector T cells correlates with loss of sustained TCR signalling and down-regulation of effector responses.
Bcl-3 is required for adjuvant mediated T cell survival and sustained inhibition of GSK3ß (87.38)
Authors: Jayaraman Padmini , Chilton Paula M , Sengupta Sadhak , Mitchell Thomas C 
Published: January 2023

Abstract
Increased transcription of bcl3 correlates with adjuvant-mediated survival of activated T cells. To determine whether Bcl3 is required for adjuvant-dependent survival of activated T cells, OT-II.Bcl3 knockout (KO) mice were challenged with ova peptide (Ova) in the presence or absence of LPS and T cell survival was assessed after 20 hr culture. Ova-treated CD4 T cells from the Bcl3 KO mice activated with or without LPS failed to survive whereas wild type (WT) OTII cells survived when activated with LPS. Using low-number adoptive transfer of WT or KO OTIIs, we show that Bcl3 is required for effective adjuvant-dependent increase in clonal expansion in response to antigen. At peak expansion, 10-fold fewer KO OTIIs were recovered after LPS treatment, compared to the WT OTIIs. Similar defects were seen in adjuvant treated OT-I.Bcl3 KO cells. GSK3ß is a target molecule whose activity is affected by adjuvants during T cell expansion. Bcl3 KO T cells do not maintain the inhibitory p(ser9) on GSK3ß as the WT OTII cells although the initial increase after T cell activation is not affected. This indicates increased phosphatase activity in Bcl3 KO T cells. Studies are underway to address unchecked phosphatase activity as a mechanism of death in the KO cells. These data establish the importance of Bcl-3 in adjuvant- mediated survival of in vivo activated T cells.
This work was supported by USPHS grants AI51377 and AI059023 and the Commonwealth of Kentucky Research Challenge Trust Fund.
Synergistic control of T cell development by Diacylglycerol kinase alpha and zeta (87.44)
Authors: Zhong Xiao-Ping , Guo Rishu 
Published: January 2023

Abstract
Signaling from the T-cell receptor (TCR) plays a critical role in thymic selections and lineage differentiation. Abnormal TCR signaling can lead to aberrant T cell development and autoimmune diseases. Thus, it is critical to understand how TCR signaling is regulated during T cell maturation. Diacylglycerol (DAG) kinases (DGKs) are a family of enzymes that catalyze phosphorylation of DAG and convert it to phosphatidic acid (PA). An important feature of DGKs mediated reaction is that both the substrate, DAG, and the product, PA, can be important second messengers. Both DAG and PA associate with and regulate the activitys of many signaling molecules. It has been hypothesized that DGKs may act as crucial regulators of receptor signaling and cellular function by modulating DAG and PA concentrations. Previous studies have demonstrated that DGKalpha and zeta are important negative regulators for peripheral T cell activation by negatively controlling DAG mediated signaling. We show here that DGKapha and zeta synergistically regulate multiple signaling pathways downstream of the TCR in developing thymocytes and that DGK activity plays a crucial role in thymic selection and CD4/CD8 lineage commitment.
A non-apoptotic FADD/caspase-8 dependent mechanism essential for T cell clonal expansion (87.48)
Authors: Walsh Craig Michael , Leverrier Sabrina , Bell Bryan D. , Arechiga Adrian F. , Weist Brian M. 
Published: January 2023

Abstract
FADD constitutes an essential component of TNF receptor-induced apoptotic signaling. Paradoxically, FADD has also been shown to be crucial for lymphocyte development and activation. Using mice expressing a T cell specific transgene encoding the death domain of FADD (FADDdd), we have found that FADD participates in T cell clonal expansion. Based on BrdU uptake assays, we have found that FADD signaling is required for transition between the late G1 and S-phase of cell cycle. The defective progression through cycle is marked by a defect in cyclin E expression and CDK2 activation. We have also identified a defect in the maintenance of S6K1 activity and ribosomal S6 phosphorylation, two essential intermediaries of proliferative signaling in T cells. We have found that caspase-8 is essential in this FADD dependent process, suggesting that FADD promotes the assembly of a non-apoptotic caspase-8 complex that serves to relieve inhibition of cell cycle progression during the G1/S transition. In line with this hypothesis, we have found that caspase-8 activity is induced in T cells in a cell-cycle dependent manner in normal T cells. The potential that this signaling axis may modulate autophagy is presently under investigation. Since TNF receptor family members play key roles in regulating adaptive immunity, these results suggest that FADD and caspase-8 serve multiple functions to maintain immune homeostasis.
From a single viral peptide epitope to effective lysis of target cells by CTL (87.50)
Authors: Sykulev Yuri , Anikeeva Nadia , Beal Allison , Vasiliver Gaia , Varma Rajat , Norris Philip J , Mattoussi Hedi , Dustin Michael L 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTL) possess a unique ability to destroy virus-infected cells by releasing cytolytic granules that upon delivery to target cells induce apoptosis. It has been proved that perhaps even a single peptide presented on a target cell renders this cell susceptible for killing by CTL. Using a novel biosensor containing quantum dots (QD) carrying controlled number of cognate and non-cognate peptide-MHC (pMHC) proteins, we have shown that pMHC attached to QD cooperate and that the presence of a single cognate pMHC with all others being non-cognate is sufficient to induce a robust CTL response.
To understand how triggering of CTL response leads to delivery of effective lethal hit to target cells, we examined granule polarization and release as well as segregation of immune receptors at interface between CTL and planar bilayer containing cognate pMHC and ICAM-1. We show that exceptionally tight ring junction formed at CTL-bilayer interface functions as a gasket precluding granule escape, which are concentrated in the secretory domain of immunological synapse. We propose that efficient target cell lysis requires 2 essential manipulations of cytolytic granules: release and effective targeting.