bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD27 co-stimulation controls tissue-specific memory CD8 T cell formation. (MUC3P.949)
Authors: Sowell Ryan , Rogozinska Magdalena , Marzo Amanda 
Published: January 2023

Abstract
Memory CD8 T cells comprise niches of circulating cells and non-circulating tissue residents. The localization of memory cells defines phenotypic and functionally distinct populations. However, the mechanisms that direct tissue-specific T cell responses remain unclear. The co-stimulatory molecule CD27 is expressed on naïve cells and memory cells in secondary lymphoid and peripheral tissues. In response to virus infection, CD27 signals enhance the magnitude of effector CD8 T cells and their capacity to respond to subsequent pathogen challenge. Moreover, CD27 may actively maintain memory CD8 T cells in secondary lymphoid tissues. We previously reported that memory cells downregulate CD27 upon entry into peripheral tissues and acquire increased lytic potential. Thus our aim was to characterize the importance of CD27 signals during the formation of tissue-resident CD8 T cells. We determined that CD27 is critical for the generation of memory cells in secondary lymphoid tissues, however it is dispensable for tissue-resident memory cells. The defect was partially dependent on mTOR because administration of rapamycin during effector differentiation rescued memory formation in absence of CD27, despite no detectable effect on the magnitude of the effector response. Analysis of CD27low cells revealed functions distict from CD27hi memory cells. Downregulation of CD27 upon entry into the periphery may be an important event that enables CD8 T cells to take up tissue residence.
Contribution of MyD88-dependent PKD1 to the development of arthritis (THER2P.954)
Authors: Yi Ae-Kyung , Yoon Tae Won , Cho Hongsik , Brand David , Kim-Hoehamer Young-In , Park Jeoung-Eun , Hasty Karen , Stuart John 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) may be initiated in part by signaling through Toll-like receptors (TLRs). Disruption of TLR signaling may provide an opportunity to halt the RA process in the earliest stages. We investigated whether protein kinase D1 (PKD1), which is essential for proinflammatory responses mediated by MyD88-dependent TLRs, plays a role in the development of experimental arthritis. The presence of TLR ligands in joint cavities induces development of reactive arthritis that is characterized by accumulation of activated inflammatory cells in the joints. Deletion of PKD1 in myeloid lineage cells prevented TLR-mediated accumulation of inflammatory cells in joints, thus protecting mice from developing reactive arthritis. Arthritis spontaneously occurring in IL-1R antagonist-deficient (IL-1rn-/-) mice was dependent on MyD88, and suppression of PKD1 expression resulted in inhibition of TLR-mediated cytokine production in IL-1rn-/- macrophages. Daily treatment with a PKD inhibitor substantially reduced the incidence and severity of arthritis in IL-1rn-/- mice. In addition, a PKD inhibitor inhibited the synergistic effect of T cell receptor and TLR2 on IL-6 production in splenocytes, and significantly reduced the incidence and severity of collagen-induced arthritis that closely mimics the pathophysiologic process of human RA. This implies that PKD1 might be one of the key factors that modulate proinflammatory responses in RA, and can be a therapeutic target for RA.
Desmoglein 3 chimeric autoantibody receptor T cells: a novel strategy for immunotherapy of pemphigus vulgaris (THER2P.958)
Authors: Ellebrecht C , Cho Michael Jeffrey , Mao Xuming , Bhoj Vijay , Tsai Ching-Yi , Nunez-Cruz Selene , Milone Michael , Payne Aimee 
Published: January 2023

Abstract
Pemphigus vulgaris (PV) is a blistering skin disease caused by autoantibodies (autoAbs) to desmoglein (Dsg) 3. PV therapy relies on general immunosuppression, risking severe infection with chronic use. Recently, chimeric antigen receptor (CAR) T cells have revolutionized immunotherapy. To evaluate this potent approach for PV, we engineered a series of Dsg3 chimeric autoAb receptors (CAARs), consisting of various Dsg3 extracellular (EC) domains fused to either T or NK receptor signaling domains. In vitro, Dsg3 CAAR T cells specifically kill target cells expressing surface anti-Dsg3 EC1, EC2, or EC3 autoAbs, with the most broad and potent cytolysis by the Dsg3 EC1-4 CAAR using the T cell CD137CD3ζ signaling domain (p=0.0001). Dsg3 CAAR T cells do not kill Fc-receptor+ cells coated with Dsg3 autoAbs or keratinocytes whose surface proteins might interact with the Dsg3 CAAR. TIRF microscopy reveals that autoAb binding by the Dsg3 CAAR forms supramolecular activation complexes, with segregation of the CAAR away from CD45, a regulator of T cell receptor activation. In a preclinical PV NSG mouse model, bioluminescent AK23 (anti-EC1 surface IgG+) hybridoma cells were efficiently killed by Dsg3 CAAR (p=0.0015), resulting in prolonged survival of CAAR-treated versus control mice (p=0.016) without skin toxicity. CAAR T cells represent a novel strategy for PV therapy that specifically targets autoantigen-specific B cells and that could readily be applied to other autoAb-mediated diseases.
CoenzymeQ10 suppresses Th17 cells and osteoclast differentiation and ameliorates experimental autoimmune arthritis mice (THER3P.1000)
Authors: Seo Hyeon-Beom , Cho Mi-La , Jeong Jeong-Hee 
Published: January 2023

Abstract
Coenzyme Q10 (CoQ10) is a lipid-soluble antioxidant synthesized in human body. This study aimed to investigate the effect of CoQ10 on inflammation and Th17 cell proliferation on an experimental rheumatoid arthritis (RA) mice model. CoQ10 or cotton seed oil as control was orally administrated once a day for seven weeks to mice with zymosan-induced arthritis (ZIA). Histological analysis of the joints was conducted using immunohistochemistry. Germinal center (GC) B cells, Th17 cells and Treg cells of the spleen tissue were examined by confocal microscopy staining. mRNA expression was measured by real-time PCR and protein levels were estimated by enzyme-linked immunosorbent assay (ELISA). Flow cytometric analysis (FACS) was used to evaluate Th17 cells and Treg cells. CoQ10 mitigated the severity of ZIA and decreased serum immunoglobulin concentrations. CoQ10 also reduced RANKL-induced osteoclastogenesis, inflammatory mediators and oxidant factors. Th17/Treg axis was reciprocally controlled by CoQ10 treatment. Moreover, CoQ10 treatment on normal mouse and human cells cultured in Th17 conditions decreased the number of Th17 cells and enhanced the number of Treg cells. CoQ10 alleviates arthritis in mice with ZIA declining inflammation, Th17 cells and osteoclast differentiation. These findings suggest that CoQ10 can be a potential therapeutic substance for RA.
Unique potential of 4-1BB agonist antibody to promote durable regression of HPV+ tumors when combined with an E6/E7 peptide vaccine (TUM2P.1016)
Authors: Bartkowiak Todd , Singh Shailbala , Yang Guojun , Galvan Gloria , Allison James , Sastry Jagannadha , Curran Michael 
Published: January 2023

Abstract
Antibody modulation of T cell co-inhibitory (e.g. CTLA-4) or co-stimulatory (e.g. 4-1BB) receptors promotes clinical responses to a variety of cancers. Therapeutic cancer vaccination, in contrast, has produced limited clinical benefit. The E6 and E7 onco-proteins of human papilloma virus drive the majority of genital and oropharyngeal cancers. We discovered 15-19 amino acid peptides from HPV16 E6/E7 for which T cell immunity correlates with disease-free survival in patients treated for high-grade cervical neoplasia. We report that intra-nasal vaccination with these peptides and the adjuvant alpha-galactosylceramide elicits systemic and mucosal T cell responses leading to reduced HPV+ TC-1 tumor growth and prolonged survival in mice. We hypothesized that the inability of these T cells to fully reject established tumors resulted from suppression in the tumor which could be ameliorated through checkpoint modulation. Combining this vaccine with checkpoint blockade produced only modest benefit; however, co-administration with a 4-1BB agonist antibody promoted durable regression of 100% of established genital HPV+ tumors. The vaccine/α4-1BB combination promoted the highest CD8+ versus regulatory FoxP3+ T cell ratios, elicited much higher infiltration by E7-specific CTL, and evoked higher densities of highly cytotoxic TcEO CD8 (&gt;70-fold) and ThEO CD4 (&gt;17-fold) phenotype T cells. Activation of 4-1BB may have a unique capacity to potentiate therapeutic cancer vaccines.
A monomeric fab fragment capable of inducing CD3 conformational change increases T cell receptor reactivity to poorly immunogenic antigens (TUM2P.1020)
Authors: Hoffmann Michele , Parks Christopher , Hansen Michael , Rajagopalan Govindarajan , Pease Larry , Schrum Adam , Gil Pages Diana 
Published: January 2023

Abstract
TCR interactions with agonist pMHCs induce a conformational change in the tails of the associated CD3 complex (CD3Δc) that contributes to efficient TCR/CD3 triggering and productive T cell responses. In contrast, TCR engagement by antagonistic pMHCs does not induce either CD3Δc or full TCR/CD3 signaling in mature T cells. Here, we have used a monovalent Fab fragment (Mono-Fab) specific for the murine CD3 complex to provide induction of CD3Δc to mature T cells engaging pMHC ligands. This Mono-Fab does not interfere with TCR capacity to bind pMHC and fails to elicit signaling in the absence of TCR/MHC interactions. Using OT-I Tg T cells and a collection of altered peptide ligands derived of the agonist peptide OVA, we show that addition of CD3Δc to OT-I T cells using Mono-Fab enables T cell responses to antagonist ligands that fail to induce CD3Δc on their own. Moreover, using a mouse model of metastatic melanoma, we show that Mono-Fab promotes in vivo T cell dependent anti-melanoma responses that are antigen-specific. This Mono-Fab can also be combined with established therapies targeting signals 2 and 3 involved in T cell activation for a synergistic effect. Our data suggests that in-trans CD3Δc provision decreases the activation threshold of T cells. We propose that targeting the CD3 complex for the induction of CD3Δc could be exploited to achieve efficient TCR/CD3 triggering by poorly immunogenic antigens in the context of vaccination against cancer and infectious diseases.
Gamma-interferon-inducible lysosomal thiol reductase is upregulated in human melanoma (TUM2P.1038)
Authors: Hastings Karen , Nguyen Jennifer , Bernert Richard , Ko Christine , Sebastiao Noemi , Hu Chengcheng 
Published: January 2023

Abstract
T cell-mediated immunity has the ability to produce durable anti-melanoma responses resulting in improved survival of patients with advanced melanoma. Antigen presentation in the tumor microenvironment directs anti-melanoma T cell responses. Gamma-interferon-inducible lysosomal thiol reductase (GILT) is critical for MHC class II-restricted presentation of multiple melanoma antigens to CD4+ T cells. However, GILT expression in melanoma has not been defined. We evaluated GILT and MHC class II expression in human primary and metastatic melanomas and nevi using immunohistochemistry. Both GILT and MHC class II expression were increased in melanocytes of primary and metastatic melanomas compared with nevi. GILT expression was increased in antigen presenting cells of primary and metastatic melanomas compared to nevi, whereas MHC class II had equivalent high expression in antigen presenting cells of all melanocytic lesions. Keratinocytes in primary melanoma lesions had increased GILT and MHC class II expression compared with nevi. Increased GILT expression in melanocytes, antigen presenting cells and keratinocytes of melanoma demonstrates that changes in the antigen presentation pathway occur in the tumor microenvironment. GILT expression in melanoma is anticipated to result in improved presentation of melanoma antigens and more effective anti-melanoma T cell responses and may be a biomarker of immune recognition of melanoma.
Targeting deposited complement component C3d to potentiate monoclonal antibody cancer therapy (VAC3P.1057)
Authors: Skarzynski Martin , Butera Vicent , Lindorfer Margaret , Vire Berengere , Farooqui Mohammed , Rader Christoph , Taylor Ronald , Wiestner Adrian 
Published: January 2023

Abstract
Treatment-induced loss of targeted cell surface antigens through trogocytosis or internalization reduces efficacy of monoclonal antibody (mAb) therapy of cancer. However, cells that escape therapy mediated by complement-fixing mAbs carry covalently deposited complement activation fragments on their cell surfaces, in particular C3d. We hypothesized that cell-associated C3d constitutes a neoantigen that could be exploited to selectively retarget cells escaping from therapeutic mAbs. We generated an anti-C3d IgG1 human/mouse chimeric mAb specific for human C3d that is not competed by C3 in human serum. For proof of concept, we used cells from chronic lymphocytic leukemia (CLL) patients that had substantially reduced CD20 levels due to treatment with the anti-CD20 mAb ofatumumab (OFA). The chimeric anti-C3d mAb bound cell surface C3d on these CLL cells ex vivo (KD = 6.7nM), and mediated complement-dependent, and antibody-dependent cellular cytotoxicity and phagocytosis in vitro. CLL cells opsonized by C3d in vivo and reacted with the anti-C3d mAb in vitro were further C3d opsonized, resulting in an amplification loop that enhanced anti-C3d mAb binding capacity and killing of target cells. In vivo, the anti-C3d mAb was effective in reducing tumor burden in blood and spleens of NSG mice xenografted with human CLL cells. Our results identify anti-C3d mAbs as a means to specifically and potently augment the therapeutic efficacy of complement-fixing mAbs.
Mammalian target of rapamycin inhibitors are promising T cell lymphoma treatment agents that synergize with ERβ agonists in vitro but not in vivo (VAC3P.1061)
Authors: Liu Yang , Sareddy Gangadhara , Pandeswara Srilakshmi , Dao Vinh , Clark Curtis , Liu Aijie , Vadlamudi Ratna , Curiel Tyler 
Published: January 2023

Abstract
mTOR signals through 2 complexes: mTORC1 and mTORC2. Approved cancer mTOR inhibitor drugs inhibit mTORC1 and are poorly effective against carcinomas and B cell lymphomas. There is thought that dual mTORC1/2 inhibitors could have better clinical efficacy. As mTOR regulates important T cell functions, we tested mTOR inhibitors in mouse EL4 T cell lymphoma. The mTORC1 inhibitor rapamycin potently suppressed EL4 proliferation in vitro, and significantly reduced EL4 tumor growth in vivo. AZD8055, a dual mTORC1/2 inhibitor, also inhibited EL4 cell proliferation in vitro, decreased in vivo tumor burden and improved survival in EL4 challenged mice, but no better than rapamycin. EL4 tumor growth was slower in females versus males, suggesting hormonal effects. As estrogen receptor (ER)β agonists reportedly can treat lymphomas we tested them. We found that ERβ agonists altered EL4 mTOR signals and a novel ERβ/mTOR physical interaction (mTORC2&gt;&gt;mTORC1) in EL4 by GST pull-down. The ERβ agonist S-equol synergized with AZD8055 to inhibit EL4 cell proliferation in vitro but did not alter EL4 tumor growth in vivo nor boost AZD8055-mediated EL4 growth suppression or survival in vivo. EL4 tumors from AZD8055 treated mice had reduced viability and proliferation (Ki67) without affecting CD3+ T cell Ki67. AZD8055 treated ID8 ovarian cancer in vivo but rapamycin did not, suggesting T cell lymphomas could respond to mTOR inhibitors better than carcinomas. ERβ effects also deserve more study.
Infection-permissive immunity provided by NA- and M2e-based vaccines protects against influenza A virus challenge and allows the induction of heterosubtypic immunity during subsequent infections. (VAC5P.1127)
Authors: Schotsaert Michael , Ysenbaert Tine , Smet Anouk , Schepens Bert , Fiers Walter , Saelens Xavier 
Published: January 2023

Abstract
Licensed influenza vaccines protect by inducing neutralizing antibodies against the main viral surface antigens hemagglutinin (HA) and neuraminidase (NA). Influenza infection induces an antiviral T-cell response that correlates with protection against a subsequent infection with a virus of a different subtype (heterosubtypic immunity, HSI). Sterilizing immunity provided by classical vaccines however, may impair the induction of cellular HSI. We previously demonstrated in mice that vaccination with virus like particles containing the ectodomain of Matrix 2 protein (M2e-VLP) induce infection-permissive immunity that not only protects against influenza A viruses of different subtypes, but also allows the induction of cellular HSI. We could recently show that infection-permissive immunity provided by vaccination with recombinantly produced soluble tetrameric N1 NA, alone or combined with M2e-VLP, not only is protective against homologous challenge with pandemic 2009 H1N1 virus but also allows establishment of cellular HSI that protects against challenge with H3N2 virus. NA-immune animals also have an enhanced frequency of circulating functional antiviral memory CD8+ T-cells upon rechallenge with heterologous virus. In contrast, vaccination with recombinant soluble trimeric H1 HA derived from pandemic 2009 H1N1 virus induces strain-specific sterilizing immunity without infection-induced cellular HSI and fails to protect against challenge with H3N2 virus.
PTPN22 deficiency leads to clearance of chronic viral infection (VIR1P.1150)
Authors: Maine Christian , Teijaro John , Sherman Linda 
Published: January 2023

Abstract
Mutations in the phosphatase-encoding gene, PTPN22, are associated with a number of autoimmune conditions, including type I diabetes, rheumatoid arthritis and systemic lupus erythematosus. To study how PTPN22 functions in immune cells we have made extensive use of a PTPN22 deficient (KO) mouse. PTPN22 KO mice exhibit increased T cell numbers, activity and function as well as increased germinal center activity and antibody production with no autoimmune phenotype. As such we are currently investigating whether loss of PTPN22 would be of benefit to host immunity against chronic viral infections. To test this hypothesis we infected PTPN22 KO mice with the persistent lymphocytic choriomeningitis virus (LCMV) cl13 virus. LCMV cl13 infection in WT B6 mice leads to chronic infection lasting over 60 days characterized by dysfunctional ‘exhausted’ virus-specific T cells. PTPN22 KO mice were able to clear the cl13 infection whereas WT mice were not. Clearance is associated with increased viral-specific CD4 T cell cytokine production 8 days post infection, as well as increased numbers of Tfh and plasma cells. At later time points after infection, PTPN22 KO mice have increased numbers of functional CD8 T cells that produce IFNγ and TNFα, whereas CD8 cells in WT mice do not. These results suggest that inhibition of PTPN22 may prove beneficial in helping to clear chronic infection.
Longitudinal study of CD4 T cell responses following Japanese encephalitis virus vaccination (VIR2P.1165)
Authors: DeGottardi Maren , Balaji Hari , James Eddie , Mai Duy , Malhotra Uma , Kwok William 
Published: January 2023

Abstract
Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis worldwide. There are currently a number of vaccines shown to effectively prevent disease. In the United States, an alum adsorbed vaccine, which consists of a formaldehyde-inactivated, attenuated JEV strain is available. Vaccine protection has been shown to be mediated by neutralizing antibody responses. However, the CD4 T cell response associated with vaccine protection has not been fully characterized. In particular, the JEV model also presents an opportunity to study the kinetics of T cell responses elicited by a multi-dose vaccination in human subjects. In this study, we characterized the longitudinal response of JEV-specific CD4 T cells following vaccination using tetramer guided epitope mapping (TGEM), direct ex vivo MHC class II tetramer staining, and intracellular cytokine staining. A total of 67 epitopes restricted by 7 common HLA-DRB1 alleles and one DRB5 allele were identified. Vaccination elicited a robust CD4 T cell response, which was substantially boosted after the second dose of vaccine was administered. JEV-specific CD4 T cells exhibited a memory phenotype and expressed CCR4 and CXCR3. Consistent with this surface phenotype, JEV specific responses were polyfunctional, secreting IFN-g, IL-4 and sometimes IL-17a. These results indicate that JEV vaccination induces a robust, polyfunctional CD4 T cell response that is similar in magnitude to other highly effective vaccines.
Coordinated immune activation by distinct mononuclear phagocytes subtypes against the mismatch of minor antigens
Authors: Atif Shaikh M , Gibbings Sophie L , Gill Ronald G , Murphy Kenneth M , Grazia Todd , Torres Raul M , Henson Peter M , Randolph Gwendalyn J , Jakubzick Claudia 
Published: January 2023

Abstract
Recently, we identified that Batf3-dependent DCs were solely responsible for the rejection of minor antigen-mismatched grafts, and demonstrated in Batf3−/− female mice, lacking Batf3-dependent DCs, the complete acceptance of minor antigen-mismatched male cells, which are normally rejected in WT female mice. Intriguingly, this rejection occurs in the absence of PAMPs, leading to considerable uncertainty of how endogenous APC subtypes become activated and licensed to induce an adaptive immune response against male cells in female mice. Investigating this gap in knowledge is the main goal of this study. Rejection of minor antigen-mismatched grafts involves all branches of the immune system: adaptive immunity (T cells), humoral immunity (B cells) and innate immunity (DCs and antigen-presenting monocytes). Although we showed that depletion of one cell type dampens or omits the inflammatory outcome of minor antigen-mismatched graft rejection, here we show how lymphoid and myeloid cell subtypes act sequentially and in concert for the induction of minor antigen-mismatched graft rejection without the presence of PAMPs. In conclusion, we believe that understanding the mechanisms of action against minor antigen-mismatched graft rejection will also apply more broadly to other diseases such as autoimmunity and cancer, in which an immune response is elicited against self-associated minor antigens.
MHC-I molecules disulfide-linked to antigenic peptides offer a simple approach to stable MHC-I/peptide complexes
Authors: Jiang Jiansheng , Boyd Lisa F , Natarajan Kannan , Margulies David H 
Published: January 2023

Abstract
A variety of engineering approaches have been used to develop MHC/beta2m/peptide complexes with improved stability for structural studies and binding to T cell receptors and other MHC-I ligands. To generate MHC-I molecules covalently linked to antigenic peptides we considered X-ray structures of the H2-Dd molecule bound to the HIV IIIB derived peptide, P18-I10, RGPGRAFVTI. Computer analysis suggested that mutation of an H2-Dd residue to cysteine in the peptide binding groove would provide an MHC heavy chain that could be refolded with P18-I10 derived peptides substituted with cysteine at peptide position P5. Using standard E. coli expression as inclusion bodies and in vitro refolding protocols, we show that a set of cysteine-substituted peptides derived from P18-I10 including the decamer P18-C5-I10 (RGPGCAFVTI) and octamer P18-C5-V8 (RGPGCAFV) could be efficiently refolded with cys-mutated H2-Dd heavy chain and beta2-microglobulin. These molecules were analyzed by antibody binding studies and by X-ray crystallography. The P18-C5-I10/cys-mut-H2-Dd complex has a structure almost identical to that of the parental P18-I10/H2-Dd complex, while the complex with the octamer P18-C5-V8 repositions the peptide such that the C-terminal valine residue occupies the F pocket of H2-Dd. Such a strategy for producing covalently coupled MHC/peptide complexes offers a new approach to generate stable complexes for structural and imaging applications that should be widely applicable.