bluuebunny/crossref_metadata_2025_split huggingface dataset:
Utilizing dual-specific CD4+ T cells in combination with oncolytic virotherapy for treatment of cancer (P2118)
Authors: VanSeggelen Heather , Hammill Joanne , Bassett Jennifer , Rabinovich Brian , Bramson Jonathan 
Published: January 2023

Abstract
Engineering T cell specificity to direct them against tumor cells utilizing chimeric antigen receptors (CARs) has emerged as an exciting technique for adoptive T cell therapy. The effectiveness of adoptive transfer therapies can be limited by a number of factors, including ability of T cells to engraft, the degree to which they infiltrate the tumor, and the immunosuppressive nature of the tumor. Oncolytic viruses (OVs) are capable of overcoming these limitations, and can serve as effective booster vaccines. In an effort to capitalize on these features, we have generated dual-specific T cells that bear TCRs specific for the OV, and CARs specific for the tumor. Using murine tumor models, we have observed significant boosting capacity of adoptively transferred dual-specific CD4+ or CD8+ T cells to tumor-bearing hosts in response to infusion with OV. Boosting with OV also dramatically influenced the distribution of the CAR-T cells, driving the cells to proliferate throughout the body. We now seek to examine the utility of dual-specific CD4+ T cells in adoptive transfer therapies. We have found that CAR-engineered CD4+ T cells are capable of mediating anti-tumor immunity, and are investigating the combination of these cells with OV to further enhance their anti-tumor efficacy. Given the promising clinical outcomes of both CAR T cells and OVs, we believe our combinatorial approach will provide a clinically feasable strategy to maximize the therapeutic potential of both therapies.
ICOS signaling in CD8+ CTL contributes to the antitumor efficacy (P2122)
Authors: Fu Tihui , Peng Weiyi , He Qiuming , Hwu Patrick , Sharma Padmanee 
Published: January 2023

Abstract
The expression of Inducible costimulator (ICOS) on CD8+ CTL and its role in antitumor immunity remains largely unknown. To address this issue, we crossed ICOS-deficient mice with pmel-1 transgenic mice to generate pmel1/ICOS-/- mice. The loss of ICOS in pmel-1CD8+ CTL cells did not affect the expression of other T cell activation markers (e.g. CD25 and CD69) but led to a significant decrease in IFN-gamma production and killing of target tumor cells. Similarly, adoptive transfer of pmel-1 cells into ICOS-ligand knockout mice showed impaired cell proliferation and decreased expression of IFN-gamma and CD107a. Most importantly, when activated ICOS- / - pmel-1 cells were adoptively tranfered into tumor-bearing mice they displayed poor proliferation and diminished antitumor activity as compared to ICOS-intact pmel-1 cells. These results provide direct evidence for the important role of ICOS in CD8+ CTL mediated antitumor immunity and have implications in the development of novel immunotherapy for cancer by manupilation of ICOS pathway.
Combined blockade of IL-4R and IKKβ inhibits melanoma growth in vivo (P2032)
Authors: Hawkins Oriana , Horton Linda , Martin Tyesha , Ayers Gregory , Richmond Ann 
Published: January 2023

Abstract
Constitutive activation of the nuclear factor kappa B (NF-κB) is common in melanoma, promoting tumor growth and metastasis. Therefore, targeting the NF-κB pathway is a possible therapeutic option. However, NF- κB activity is critical for the development and function of immune cells. Melanoma bearing C57BL/6 mice received BMS-345541, a selective inhibitor of the inhibitor of κB kinase (IKKβ). No difference was observed in tumor size. Tumors were analyzed by flow cytometry and intracellular cytokine staining revealed an early induction of Interleukin-4 (IL-4) within the leukocyte compartment of the tumor, with drug treatment. The production of IL-4 leads to an early switch to a tumor promoting T helper 2 (Th2) driven immune response, counteracting the anti-tumor effect of IKKβ inhibition. IL-4 receptor (IL-4R) expression on human melanoma cells has been documented; however the effects of IL-4 on these cells have generated contradictory reports. In vitro IL-4 delivery did not alter either the proliferative capacity of IL-4R expressing melanoma cell lines or their sensitivity to BMS-345541. In nude mice bearing an IL-4 transduced human A375 melanoma , BMS345541 was given in combination with IL-4R blocking antibody. This combination inhibited tumor growth compared to BMS-345541 treatment alone, similar to previous studies in C57BL/6 mice. IL-4R antagonists are in clinical trials for the treatment of asthma, making the translational prospects of this study extraordinarily viable.
Human effector memory T lymphocytes express pre-existing hypoxia-adaptation genes and are resistant to hypoxia (P2036)
Authors: Xu Yang , Metelissa Leonid , Savoldo Barbara , Dotti Gianpietro 
Published: January 2023

Abstract
Hypoxia that is a hallmark of solid tumors is considered an inhibitory mechanism that hampers the clinical benefits of T-cell-based immunotherapy in cancer patients. Studies have shown indeed that circulating human peripheral blood (PB) T cells activated in hypoxia have reduced proliferation and survival. However, antigen-specific T cells generated ex vivo and adoptively transferred in cancer patients are mostly (&gt;80%) effector memory T cells (TEM) while this subset represents &lt;20% of total circulating PB T cells. How hypoxia selectively influences TEM cells has not been studied. In sharp contrast with T cells from PB, we found that proliferation, survival and cytotoxic functions of ex vivo expanded T cells are augmented in hypoxia. Importantly, we found that TEM cells directly isolated from PB show properties similar to those of expanded T cells when compared to isolated naïve and central memory cells, suggesting that the resistance to hypoxia is an intrinsic property of TEM. Finally, resistance to hypoxia is associated with an increased hypoxia-adaptation genes. Expression of HIF-1a and downstream glycolytic genes are indeed higher in TEM cells, suggesting a genetic “pre-adaptation status” of TEM to hypoxia. Together, we show that primary TEM and ex vivo generated antigen-specific T cells are resistant to hypoxia, and this finding has relevant implication for the clinical translation of adoptive immunotherapy in cancer patients.
Nardilysin-dependent proteolysis of cell-associated VTCN1: A novel pathway and biomarker of type 1 diabetes development. (P4117)
Authors: Savinov Alexei , Radichev Ilian , Maneva-Radicheva Lilia , Amatya Christina , Parker Camille , Ellefson Jacob , Wasserfall Clive , Atkinson Mark , Burn Paul 
Published: January 2023

Abstract
Hyper-active T cell responses towards pancreatic β-cells highlight type 1 diabetes (T1D). V-set domain-containing T cell activation inhibitor-1 (VTCN1) protein, a B7-like negative co-stimulatory molecule, acts inhibiting T cell activation. Here, we unveil the general defect within VTCN1 pathway, common for diabetes-prone mice and subset of T1D patients. Specifically, gradual loss of VTCN1 from antigen-presenting cells and pancreatic islets progressed alongside natural T1D development and compromised VTCN1 functionality, potentiating hyper-proliferation of diabetogenic T cells. Mechanistically, loss of membrane-tethered VTCN1 relied on proteolytic cleavage mediated by the metalloproteinase nardilysin (NRD1). Cleaved soluble VTCN1 fragment (sVTCN1) was detected at high levels in peripheral blood of 47% of pediatric T1D patients, compared to 5% of healthy subjects. Blood sVTCN1 levels correlated with the disease aggressiveness and duration, highlighting potential use of sVTCN1 as the new T1D biomarker. Discovered VTCN1-NRD1 axis identifies NRD1 as a novel target for future anti-diabetic therapeutics.
Chronic inflammatory liver disease in mice expressing a CD28-specific ligand (P4099)
Authors: Corse Emily , Gottschalk Rachel , Park Joon Seok , Sepulveda Manuel , Loke P'ng , Sullivan Timothy , Johnson Linda , Allison James 
Published: January 2023

Abstract
Inflammation of the normally tolerant liver microenvironment precedes the development of chronic liver disease. Study of the pathogenesis of autoimmune liver diseases, such as autoimmune hepatitis (AIH), has been hampered by a lack of autochthonous chronic animal models. Through our studies of T cell costimulation, we generated transgenic mice expressing a ligand specific for the CD28 receptor, which normally shares ligands with the related inhibitory receptor CTLA-4. The mice spontaneously develop chronic inflammatory liver disease with several pathologies found in AIH, including elevated serum aminotransferases in the context of normal alkaline phosphatase and bilirubin levels, lymphocytic inflammation, focal necrosis, oval cell hyperplasia, and fibrosis. The prevalence of IFN-γ-producing CD8(+) T cells in the livers of transgenic mice suggests a role for autoimmune cytotoxicity in the chronic disease state. The CD28 ligand-specific transgenic mice will facilitate evaluation of CD8(+) T cell function in liver disease pathologies found in AIH.
Regulation of pathogenic GL7-expressing helper T cells by Tregs and CD137 (P4111)
Authors: Gauld Stephen , Leonardo Steven , De Santis Jessica 
Published: January 2023

Abstract
Dissecting the mechanisms that support autoantibody production is essential to our understanding of autoimmune diseases such as lupus. Recent work from our laboratory has shown that regulatory T cells (Tregs) play a key role in suppressing autoantibody production by helping to maintain B cell anergy. Our long-term goal is to understand how Tregs regulate B cell anergy. Our studies indicate that helper T cells play a key role as an intermediate between Treg and anergic B cells. Specifically, the absence of Tregs rapidly (within 4 days) drives the expansion of CD4+GL7+ helper T cells. This gross population can be further divided into 2 populations: germinal center follicular T cells (GC-TFH) and a novel non-TFH subset termed TAN cells. These GL7+ helper T cell subsets deliver potent signals to naïve and anergic B cells, inducing significant antibody production from both populations. Recent studies have begun to identify the signals that drive the expansion of GC-TFH and TAN cells in the absence of Tregs. We have shown that treatment of Treg-deficient mice with an agonistic anti-CD137 antibody reduces the expansion of CD4+GL7+ helper T cells (both GC-TFH and TAN cells). This treatment also reduced the expansion of class-switched B cells and abolished autoantibody production. Collectively, our studies indicate that CD137 signaling plays a key role in the expansion of pathogenic helper T cell subsets that support autoimmunity in the absence of Tregs.
Qa-1b has antigen presentation and immunoregulatory roles during aerogenic Mycobacterium tuberculosis infection (P3296)
Authors: Bian Yao , Shang Shaobin , Shanmuganad Sharmila , Siddiqui Sarah , Wang Chyung-Ru 
Published: January 2023

Abstract
The mouse MHC Ib molecule Qa-1b plays roles in both antigen presentation and immune regulation in different disease models. Further, the human homologue of Qa-1b, HLA-E, can present Mycobacterium tuberculosis (Mtb) peptides to CD8+ T cells. In viral infections, Qa-1b suppresses NK and CD8+ T cell function by interacting with inhibitory CD94/NKG2A molecules. In addition, Qa-1b can activate CD8+ T regulatory cells, which suppress CD4+ T cell responses in EAE. However, the role of Qa-1b in Mycobacterium tuberculosis (Mtb) infection is unknown. In this study, we find Qa-1b expression is upregulated in a mouse model of low-dose, aerogenic infection with Mtb. We find that Qa-1b, like HLA-E, can bind a number of Mtb peptides; a subset of these peptides are recognized by CD8+ T cells and induce IFN-γ production during aerogenic Mtb infection. To determine the overall contribution of Qa-1b to Mtb infection, we infected Qa-1b-deficient and wild-type mice. Qa-1b-deficient mice had increased production of pro-inflammatory cytokines compared to wild-type, indicating Qa-1b plays a regulatory role in Mtb infection. Infected Qa-1b-deficient mice also show increased expression of CD94/NKG2A on CD8+ T cells, indicating a potential mechanism by which Qa-1b controls immune responses in Mtb infection. Our data suggest that during Mtb infection, Qa-1b is uniquely able to both activate immune responses through antigen presentation as well as suppress immune responses via regulatory mechanisms.
T cell-intrinsic MyD88 signaling regulates Th1 polarization and IFN-γ mediated immunopathology (P3316)
Authors: Yarovinsky Felix 
Published: January 2023

Abstract
Activation of Toll-like receptors (TLRs) by pathogens triggers cytokine production and T cell activation, immune defense mechanisms that can cause immunopathology. Previous studies have established that Myd88-/- mice, which cannot signal through IL-1-family receptors and most TLRs, including TLR11, are acutely susceptible to infection with a protozoan parasite Toxoplasma gondii. Myd88-/- mice fail to activate T cells and produce IFN-γ essential for host resistance to the parasite. Using a comprehensive panel of cell type-specific MyD88-deficient mice, we show that T cell-intrinsic MyD88 signaling, rather than MyD88 activation in antigen-presenting cells, had a major role in Th1 polarization. T cell-specific elimination of MyD88 reduced IFN-γ secretion by CD4+ T cells. We also show that IFN-γ production by CD4+ Th1 cells during mucosal responses to T. gondii resulted in dysbiosis and the elimination of Paneth cells. Paneth cell death led to loss of antimicrobial peptides and occurred in conjunction with uncontrolled expansion of the Enterobacteriaceae family of Gram-negative bacteria. The expanded intestinal bacteria were responsible for the parasite-induced intestinal pathology. The investigation of cell type-specific factors regulating Th1 polarization during T. gondii infection identified the T cell-intrinsic TLR pathway as a major regulator of IFN-γ production in CD4+ T cells responsible for Paneth cell death, dysbiosis and intestinal immunopathology.
The inhibition of allergic airway inflammation by regulatory T cells induced with Trichinella spiralis infection. (P3322)
Authors: Kang Shin Ae , Cho Min Kyoung , Park Mi Kyung , Yu Hak Sun 
Published: January 2023

Abstract
In several recent reports, helminthic infections could reduce various allergic response and immune response. In previous study, we demonstrated that Trichinella spiralis infection could ameliorate allergic airway inflammation and CD4+CD25+ Foxp3+ T (Treg) cells were recruited in the lung. In order to know function of recruited T reg cell of T. spiralis infected mice, we elicited allergic airway inflammation with OVA-alum and injected Treg cell by intra-venous which was isolated from T. spiralis infected mice. Treg cells induced with T. spiralis infection inhibit OVA-induced airway inflammation. A significant reduction of immune cell infiltration and the concentrations of IL-4 and IL-13 were lower than those of control. The gene expression levels of various important homing receptors and activated Treg marker in Treg cells of each group were analyzed via real-time PCR. As a result, the expression of CTLA-4, CCR5 and CCR9 was significantly increased in Treg cells by T. spiralis infection. These data identify Treg cells induced by T. spiralis infection as capable of regulating allergic airway responses by migration to inflamed tissues and activation.
Identification of PE/PPE epitopes of Mycobacterium leprae recognized by peripheral blood mononuclear cells in leprosy (P3332)
Authors: Choi Aaron , Legaspi Anna , Hou Wanqiu , Modlin Robert 
Published: January 2023

Abstract
Leprosy, a chronic disease caused by the intracellular pathogen Mycobacterium leprae (M leprae), is a major health concern. Mycobacterial PE/PPE proteins may play a role in the interface of host-pathogen interaction and disease development. However, it remains unclear how the immune system recognizes those proteins and their components. Here, we have employed an Internet online-based tool called immune epitope database analysis resource (http://tools.iedb.org) to predict and analyze epitopes of M. leprae PE/PPE proteins. Thirteen candidate T cell epitopes were identified with a potential to bind a broad range of MHC class II haplotypes. Using ELISA, we found that 7/13 of predicted epitopes stimulated IFN-γ production in peripheral blood mononuclear cells (PBMCs) of tuberculoid or reversal reaction leprosy patients but not healthy donors. Among them, ML0588 (7-21) exhibited the broadest stimulatory capacity, eliciting IFN-γ production by PBMCs in 4/7 patients. Taken together, we have utilized the immune epitope database to identify seven peptides encoded by M. leprae PE/PPE proteins as strong candidate T cell epitopes for the induction of IFN-γ production in leprosy patients. These findings suggest that T cell recognition of these proteins may contribute to host defense against leprosy and be of potential use in a diagnostic test for leprosy.
The role of LIGHT a member of the TNFSF in regulation of secondary immunity to Leishmania major (P3344)
Authors: Okwor Ifeoma , Xu Giu-Lian , Uzonna Jude 
Published: January 2023

Abstract
We previously showed that LIGHT, (lymphotoxin like, exhibits inducible expression and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes) and its receptors are critically important for initiation of CD4+ T helper (Th1) cell response and development of primary immunity to Leishmania major infection in mice. Here, we further characterized the contributions of this molecule in the activation of dendritic cells (DCs), L. major-induced IL-12 production and secondary anti-Leishmania immunity. We show that blockade of LIGHT significantly impaired DC maturation and expression of costimulatory molecules including, CD40 and CD86. In addition, blockade of LIGHT signaling lead to impairment in L. major-induced IL-12p40 production by DCs in vivo and a concomitant reduction in the number of cells recruited into the draining lymph nodes as well as early IFN-γ production by CD4+ T cells following L. major infection. Furthermore, we show that LIGHT is required at both priming and maintenance stages of anti-Leishmania immunity. Interestingly, blockade of LIGHT had no effect on delayed-type hypersensitivity (DTH) response and control of parasite burden following secondary L. major challenge. Collectively, these results show that LIGHT is critical for IL-12 production by DCs and subsequent priming and maintenance of IFN-γ response by CD4+ T cells during primary but is dispensable during secondary anti-Leishmania immunity.
Production of monoclonal camelid antibodies by means of hybridoma technology (P3376)
Authors: Holzloehner Pamela , Schliebs Erik , Maier Natalia , Füner Jonas , Micheel Burkhard , Heilmann Katja 
Published: January 2023

Abstract
Here we describe the first monoclonal immunoglobulins produced by camelid hybridomas. Hybridomas were generated by using the CBF7 heteromyeloma line to fuse peripheral blood lymphocytes from New World camels (llamas, alpacas) and Old World camels (Bactrian camels). The hybridomas produced immunoglobulins of IgG1, IgG2 and IgG3 subclasses as defined by both enzyme immunoassays and PCR and subsequent gene sequencing. The subclass-specific murine monoclonal antibodies were also produced in the experiments described here. They could be used to detect conventional four-chain camelid antibodies (of subclass IgG1) and heavy chain-only antibodies (both of subclasses IgG2 and IgG3). The different monoclonal heavy chain-only immunoglobulins and the producing hybridomas are of special interest since they can be used to produce further secondary reagents (murine monoclonal antibodies) for the characterization of different camelid antibodies and to solve some as yet unresolved problems in the molecular biology of heavy chain antibody-producing cells. Most importantly, the technique described here can be the basis for the production of full-length heavy chain-only monoclonal antibodies of camelids directed against specific antigens.
Multiplex gene expression analysis for low abundance intracellular mRNAs in individual cells by RNA flow cytometry (P3381)
Authors: Park Emily , Lomas Woodraw , Hanley Mary , Mittar Dev , Su Nan , Luo Yuling , Maino Vernon 
Published: January 2023

Abstract
A variety of RNA analysis technologies is available for the detection of RNA transcripts from bulk cell populations. However, the techniques for RNA detection from single cells have been limited due to lack of sensitivity in detecting specific mRNAs present at low abundance in individual cells. By designing novel target-specific probes and adapting an in situ signal amplification method based on the RNAscope® detection platform, we demonstrate the specific and sensitive detection of intracellular RNAs in single cells by flow cytometry (RNAFlow). This method had sufficient sensitivity to distinguish cells containing low abundance RNA transcript. Furthermore, multiple distinct RNA targets were simultaneously detected with a high specificity in single cells without interference. The method can quantify the frequency of cells expressing specific RNA as well as the number of RNA copies in each expression-positive cell. We will present quantitative data to characterize RNA target-specific signal detection, demonstrated in applications such as HIV infection and immune activation. The RNAFlow assay, independently or as combined assay with co-protein detection, represents a valuable research tool for the specific and sensitive detection of multiple RNA transcripts in individual cells in heterogeneous biological specimens. Overall, this method will be useful for the analysis of functionally important RNA species from single cells, even at very low copy numbers.
Simplified T & B lymphocyte enumeration on a novel cell analyzer (P3390)
Authors: Clor Julie , Gillis Katherine , Tyagarajan Kamala 
Published: January 2023

Abstract
Characterization and enumeration of T and B cell lymphocyte sub-populations is crucial in immunological research applications for understanding of immune status and response. This analysis has typically been performed using sophisticated single platform flow cytometry or double platforms, to which many researchers may have limited access. In this study, we evaluated performance of a novel, low-cost, small footprint platform, the Muse™ Cell Analyzer which is based on microcapillary cytometry and utilizes guided touchscreens, for immunophenotyping analysis. Whole blood and peripheral mononuclear blood cell samples a were analyzed using simplified, no wash, novel two color strategies and results for CD4 T cells, CD8 T Cells and B cells count and percentage of lymphocytes were compared against well-established predicate methods. Comparison of results demonstrates good correlations and low bias for each strategy. Precise results were obtained for all assays tested as shown by the low coefficient of variation (CV&lt;10%). Low operator to operator variability with inter-operator CV’s of &lt;10% was also observed for the analysis. Further, T or B lymphocyte characterization on samples is performed using low number of cells. The accurate and precise enumeration and evaluation of T and B lymphocytes in a simple, accessible system can empower researchers to obtain information on critical T& B lymphocyte populations of interest with ease.