bluuebunny/crossref_metadata_2025_split huggingface dataset:
Homogeneity of the adaptive immune repertoire of primary and metastatic ovarian tumors (P2205)
Authors: Sherwood Anna , Emerson Ryan , Desmarais Cindy , Tewari Muneesh , Guenthoer Jamie , Carlson Chris , Drescher Charles , Robins Harlan 
Published: January 2023

Abstract
The presence, abundance, population, and diversity of Tumor Infiltrating Lymphocytes (TILs) have been identified as prognostic indicators in several cancers. Emerging cancer therapeutics including immunomodulators and adoptive T-cell therapy highlight the need to better understand and track this population of T cells. We have developed a multiplex PCR amplification assay for the rearranged T-cell receptor Beta (TCRB), followed by deep sequencing to characterize the adaptive immune response spatially within ovarian tumor. Specifically, we used our assay to characterize the heterogeneity of ovarian tumor TIL populations. We sequenced 10 primary and metastatic ovarian tumors collected from 5 patients. Each tumor was divided into a grid pattern with 8-22 sections (tumor size dependent). For each tumor section we collected data on the number, diversity, and the unique CDR3 sequences carried by the TILs, to characterize the intra-tumor heterogeneity of TIL count, diversity, and T cell clone overlap. In addition, we sequenced the immune repertoire from a paired peripheral blood sample to identify if the immune response was tumor specific. We found that the immune response within peripheral blood was significant and quantifiably different than the tumor repertoire. TIL repertoire within sections of a tumor showed high similarity to each other. These data indicate that the TCR repertoire of the ovarian tumor environment is tumor specific and homogeneous.
NLRP3 inflammasome independent release of IL-1β by tumor cell lysates stimulated dendritic cells (P2215)
Authors: Wang Minan , Gollnick Sandra 
Published: January 2023

Abstract
Photodynamic therapy (PDT) has long been known for capable of killing tumor cells and induces anti-tumor immunity. The mechanism of how PDT can enhance anti-tumor response is not clear. We tested the hypothesis that bone marrow derived dendritic cells (BMDCs) activated by PDT-generated tumor cell lysates (PDTTCL) can induce anti-tumor response in Lewis lung carcinoma (LLC), a poorly immunogenic tumor model. We found that LLC tumor growth was retarded by immunization with BMDCs pulsed by PDTTCL. Furthermore, therapeutic effects are observed in preexisting LLC tumor when PDT is combined with PDTTCL treated BMDCs and R848 (TLR7/8 ligand) therapy. This study aims to understand how PDT enhances tumor immunogenicity and provides insight to improve both in situ PDT and PDT vaccine efficacy in combating secondary disease. We found that stimulation of BMDCs with PDTTCL leads to dramatic increase in pro-inflammatory cytokine production, including IL-1β, which plays a critical role in PDT efficacy. Previous studies have shown that production and activation of IL-1β require both TLR signaling and activation of NLRP3 inflammasome. IL-1β production induced by PDTTCL in BMDCs is MyD88 dependent, but the activation of IL-1β appears to be dependent upon elastase and proteinase 3 rather than NLRP3 inflammasome. Together, our findings reveal a novel mechanism of IL-1β activation in BMDCs that does not require NLRP3 inflammasome in response to danger signals from tumor cell lysates.
Withaferin A, a potent and abundant component of Withania somnifera root extract, reduces myeloid-derived suppressor cell function (P2103)
Authors: Sinha Pratima , Ostrand-Rosenberg Suzanne 
Published: January 2023

Abstract
Myeloid cells play a crucial role in growth and metastasis of malignant tumors. Tumor infiltrating myeloid cells include myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages. These cells infiltrate into tumor and suppress tumor immunity by their inherent immune suppressive activity which is enhanced by interactions with each other (cross-talk). The root extract of the plant Withania somnifera (WRE) has been reported to reduce tumor cell proliferation and angiogenesis. We hypothesize that WRE or its constituents impact tumor infiltrating myeloid cells and thereby boost anti-tumor immunity. HPLC and MS analysis revealed that Withaferin A (WA) is the most abundant constituent of WRE. A prominent effect of MDSC is their production of IL-10 which increases upon cross-talk with macrophages, promoting type-2 immunity. WA reduces constitutive and cross-talk induced IL-10 secretion from MDSC. Macrophage secretion of IL-6 and TNFa cytokines that are characteristic of M1-type macrophages and that also increase MDSC accumulation and function, are also reduced by WA. Much of the T cell suppressive activity of MDSC is due to MDSC production of reactive oxygen species (ROS), and WA significantly reduces MDSC production of ROS. Thus adjunctive treatment with WA has the potential to concomitantly reduce myeloid cell mediated immune suppression, to polarize immunity towards a tumor-rejecting type I phenotype, and to facilitate the development of anti-tumor immunity.
IFN-γ activates p300 to over-ride silencing effects of DNA methylation to activate GPR109A and SLC5A8 transcription (P2105)
Authors: Bardhan Kankana , Martin Pamela , Browning Darren , Ganapathy Vadivel , Robertson Keith , Liles Georgia , Lee Jeffery , Liu Kebin 
Published: January 2023

Abstract
Short-chain fatty acids, metabolites produced by colonic microbiota from fermentation of dietary fiber, act as anti-inflammatory agents in the intestinal tract to suppress inflammation-mediated diseases such as inflammatory bowel disease and colorectal cancer. GPR109A and SLC5A8, receptor and transporter, respectively, for short chain fatty acids, are often silenced in human colon carcinoma cells. We demonstrated that the GPR109A and SLC5A8 promoter DNA is hypermethylated in human colon carcinoma cells in vitro and in vivo. Strikingly, IFN-γ, a cytokine secreted by activated cytotoxic T lymphocytes, effectively activated GPR109A and SLC5A8 transcription without altering their promoter DNA methylation levels. Analysis of the IFN-γ signaling pathways indicated that IFN-γ-activated pSTAT1 binds to the promoter of immediate early target gene p300 to rapidly activate p300 transcription. p300 then binds to the GPR109A and SLC5A8 promoters and induces chromatin hyperacetylation, resulting in chromatin remodeling in the hypermethylated GPR109A and SLC5A8 promoter regions. pSTAT1 then directly binds to the remodeled but still hypermethylated GPR109A and SLC5A8 promoters to activate transcription of secondary response genes GPR109A and SLC5A8. Taken together, our data suggest that the host immune system might use IFN-γ to counteract DNA methylation-mediated silencing of tumor suppressor genes as a mechanism to suppress tumor development.
Cytolytic capacity of KIR2DS1+ decidual natural killer cells in the fetal-maternal interface (P3203)
Authors: Crespo Angela , Tilburgs Tamara , Strominger Jack 
Published: January 2023

Abstract
Decidual natural killer cells (dNK) are the most abundant lymphocytes at the fetal-maternal interface. dNK have been shown to be less cytotoxic but to secrete more cytokines and growth factors that may help trophoblast invasion than peripheral blood NK cells (pNK). Previous studies have shown that mothers expressing the activating Killer Immunoglobulin-like Receptor-S1 (KIR2DS1) are protected from pregnancy complications when carrying a fetus expressing more HLA-C2 group alleles than them (Hiby et al, 2010). However, no experimental studies have addressed the mechanisms involved in this beneficial effect. In this study, we demonstrate that a significantly higher percentage of dNK express KIR2DS1 and KIR2DL1 than pNK cells. dNK and pNK express similar levels of perforin and granzyme B but dNK contain increased granulysin levels. Although pNK are more cytotoxic than dNK against HLA negative targets, the total pNK cytotoxicity is efficiently inhibited by HLA-C1+ and HLA-C2+ targets. In contrast, HLA-C2+ targets only inhibit the cytotoxicity of total dNK from KIR2DS1- but not KIR2DS1+ mothers. Assessment of single KIR2DL1+ and KIR2DS1+ pNK and dNK shows that HLA-C2 efficiently inhibits KIR2DL1+ but not KIR2DS1+ cell cytotoxicity. These results suggest a dominant role for KIR2DS1+ cells in the total dNK population. Future studies will focus on the influence of the fetal/maternal HLA-C genotype on KIR2DS1+ cell function and the identification of cytokines secreted by KIR2DS1+ dNK.
Antigen persistence augments maintenance of effector-like non-lymphoid T cells but may impair protective efficacy (P3207)
Authors: Beura Lalit , Anderson Kristin , Schenkel Jason , Locquiao Jeremiah , Fraser Katy , Vezys Vaiva , Masopust David 
Published: January 2023

Abstract
The innovative concept of developing persistent vaccines is predicated on the hypothesis that tonic antigen stimulation may maintain abundant effector-like memory CD8 T cells in tissues. We wished to test the impact of persistent antigen stimulation on tissue localized CD8 T cell distribution, differentiation, and function in a reductionist model. To this end, we compared T cell responses in mice infected with strains of LCMV that cause acute versus persistent infections. In contrast to previous reports in this model, persistent infection resulted in the maintenance of ~30 fold more virus-specific lung CD8 T cells relative to acute infection. This interpretation was dependent on the use of refined in vivo labeling techniques that allowed discrimination between T cells present within tissue parenchyma and abundant populations that fail to be removed from vascular compartments despite perfusion. Persistent infection also maintained higher granzyme B expression in lung and greater accumulation of virus-specific T cells in the large intestine, liver, kidney, and female reproductive tract. Surprisingly, quantity and granzyme B expression did not correlate with protective capacity against a local respiratory infectious challenge. In fact, memory CD8 T cells established via acute infection afforded better protection. These data illustrate that protective efficacy cannot be predicted based solely on effector-like CD8 T cell quantity in tissues and phenotype.
Defective induction of regulatory B cells by decidua stromal cells underlies the pathogenesis of preterm birth (P3213)
Authors: Wang Zhao-Yuan , Mathiarasu Aditya , Hong Peng , Xu Yi , Li Aiqun , Bluth Martin , Lum Lawrence , Huang Bihui , Romero Roberto , Cerutti Andrea , Chen Kang 
Published: January 2023

Abstract
Preterm birth is the leading cause of mortality and morbidity of newborns. Many maternal autoimmune conditions that predispose to preterm birth involve the dysregulation of autoreactive B cells that contribute to disease pathogenesis. Despite the fundamental roles of B cells as effectors and regulators of immunity, their functions in pregnancy are poorly understood. We found that human B cells undergo activation, class switching, memory and plasmacytoid differentiation in the decidua in situ. Preterm decidua harbors increased numbers of total B cells and B-1 cells but diminished numbers of IL-10-producing regulatory B cells in vivo. Furthermore, stromal cells in preterm decidua, as compared to those in term decidua, stimulate increased B cell activation and the differentiation of potentially autoreactive B cells, but are defective in inducing IL-10 expression by B cells. These results demonstrate that preterm birth involves abnormal B cell functions at the maternal-fetal interface, whereby decidua stromal dysfunction engenders unique populations of B cells with increased autoreactivity but diminished regulatory functions, and suggest that regulatory B cells are important for the maintenance of normal pregnancy. These studies provide insights into the functions of B cells at the maternal-fetal interface during pregnancy and inform the design of B cell-targeted therapies to treat preterm birth and other pregnancy complications involving B cell dysfunction.
Persistent enteric murine norovirus infection is associated with functionally suboptimal virus-specific CD8 T cell responses (P3187)
Authors: Tomov Vesselin , Osborne Lisa , Dolfi Douglas , Sonnenberg Gregory , Monticelli Laurel , Mansfield Kathleen , Virgin Herbert , Artis David , Wherry E. 
Published: January 2023

Abstract
Norovirus (NV) gastroenteritis is a major contributor to global morbidity and mortality, yet little is known about immune mechanisms leading to NV control. Previous studies using the murine norovirus (MNV) model have established a key role for T cells in MNV clearance. Despite these advances, important questions remain regarding the magnitude, location, and dynamics of the MNV-specific T cell response. To address these questions, we identified MNV-specific MHC class I immunodominant epitopes by conducting an overlapping peptide screen. Using MHC class I-peptide tetramers, we tracked MNV-specific CD8 T cells in lymphoid and mucosal sites during infection with two strains with distinct biological behavior: acutely-cleared CW3 and persistent CR6. Here, we show that enteric MNV infection elicited robust T cell responses primarily in the intestinal mucosa, and that MNV-specific T cells dynamically regulated the expression of surface molecules associated with activation, differentiation, and homing. Furthermore, compared to MNV-CW3, chronic infection with MNV-CR6 resulted in fewer and less functional CD8 T cells. Finally, MNV-specific CD8 T cells were capable of reducing viral load in persistently infected Rag1-/- mice, consistent with a crucial role for these cells in vivo. Collectively, these data provide fundamental new insights into the adaptive immune response to NV and bring us closer to understanding the correlates of protective antiviral immunity in the intestine.
GM-CSF control of macrophage differentiation is a critical effector of Th17 responses in vivo (P4180)
Authors: Barin Jobert , Baldeviano G. , Talor Monica , Stickel Natalie , Bedja Djahida , Gabrielson Kathleen , Rose Noel , Cihakova Daniela 
Published: January 2023

Abstract
Like many models of tissue-localized autoimmune disease, experimental autoimmune myocarditis (EAM) is mediated by Th17-directed pathology. The composition of cardiac infiltrates in EAM is predominantly monocytic, indicating that these cells are likely to be major inflammatory effectors in this disease process. In common with other Th17 autoimmune diseases, IFNγ-deficient mice develop very severe disease. We observed enhanced GM-CSF production from autoantigen-specific IFNγ-/- CD4+ cells, derived from the heart or spleen, suggesting that the induction of GM-CSF by Th17 cells controls the involvement of monocytes and macrophages in disease. After generating IFNγ-/-IL17A-/- double-knockout mice, we determined that the production of GM-CSF in CD4+ is driven by IL17 signaling. Towards this end, we generated bone marrow-derived macrophages (BMMϕ) in vitro, in the presence of GM-CSF, rather than M-CSF. Following adoptive transfer, recipients of GM-CSF BMMϕ developed more severe disease than controls. This severe disease was accompanied by enhanced fibrosis and compromised cardiac function. This novel report links GM-CSF control of macrophage differentiation to the functional consequences of Th17-mediated autoimmunity.
Identification of posttranslational modified autoantigens in Type 1 diabetes. (P4152)
Authors: Yang Mei-Ling , Connolly Sean , Wen Li , Herold Kevan , Mamula Mark 
Published: January 2023

Abstract
Autoimmune diseases such as lupus, rheumatoid arthritis, multiple sclerosis, and type 1 diabetes (T1D) frequently target self proteins that undergo posttranslational protein modifications. Oxidative stress, which is implicated in the pathogenesis of T1D, leads to various protein modifications including isoaspartyl (isoAsp) and carbonyl formation. We have previously identified that isoAsp protein modification can trigger T cell autoimmunity in both human and murine models of lupus. However, the role of oxidative stress-induced posttranslational modifications in T1D is still poorly understood. Herein, we demonstrate that isoAsp modification and oxidative carbonylation accumulate in pancreas and/or pancreatic islet proteins from spontaneous diabetes-prone NOD mice. Specifically, isoAsp content were elevated in both target organ (pancreas) and peripheral erythrocytes of diabetic NOD mice compared to pre-diabetic NOD mice. Moreover, we identified the beta subunit of prolyl-4-hydroxylase (P4hb) as a protein that is abundantly carbonylated in pancreatic islet. Sera from NOD mice and early onset diabetic patients contain antibodies that recognized isoAsp- and/or carbonyl-modified pancreas proteins including P4hb protein. These studies provide a novel insight into the mechanism of breaking immune tolerance and the diagnostic and/or therapeutic targets associated with isoAsp and carbonylated posttranslational protein modifications in T1D.
The critical role of Th1 cytokine polarization in murine primary biliary cirrhosis. (P4154)
Authors: Tsuda Masanobu , Zhang Weici , Yang Guo-Xiang , Ando Yugo , Kawata Kazuhito , Tanaka Hajime , Leung Patrick , Lian Zhe-Xiong , Gershwin M. Eric 
Published: January 2023

Abstract
Primary biliary cirrhosis (PBC) is a female predominant organ specific autoimmune disease with specific destruction of intrahepatic small biliary epithelial cells (BECs). Patients develop a multilineage response to the E2 component of pyruvate dehydrogenase (PDC-E2) involving autoantibody. We previously reported that IL-12p40 deficiency eliminated biliary disease in dnTGFβRII mice, suggesting that IL-12 and IL-23 are important in the development of biliary disease. IL-12 induced Th1 and IL-23 induces Th17, which express both IFN-\#933; and IL-17, but the role of IL-12p40 in PBC remains unidentified. To further investigate the role of the IL-12p40 in this murine model of autoimmune biliary disease, we immunized IL-12p40-/-, IFN-\#933;-/- and IL-17-/- mice with a xenobiotic mimic of PDC-E2 , 2-octynoic acid (2-OA), coupled to bovine serum albumin. The PBC-related autoepitope of PDC-E2 contains lipoic acid, and previous work has demonstrated that mimics of lipoic acid following immunization of mice lead to a PBC-like disease. Importantly IL-12p40-/- and IFN-\#933;-/- mice immunized with 2-OA did not manifest autoimmune biliary disease. In contrast to IL-12p40-/- and IFN-\#933;-/- mice, IL-17-/- mice developed autoimmune biliary disease. These data suggest that signaling by way of Th1 polarizaion via IL-12p40 is an essential requirement for the development of autoimmune cholangitis. These data have implications for the biologic therapy of patients with primary biliary cirrhosis.
The development of tertiary lymphoid organs in the central nervous system facilitates determinant spreading of the MP4-specific T cell response (P4164)
Authors: Kuerten Stefanie , Schickel Achim , Kerkloh Christian , Recks Mascha , Addicks Klaus , Ruddle Nancy , Lehmann Paul 
Published: January 2023

Abstract
Accumulating evidence indicates that B cells aggregate into tertiary lymphoid organs (TLOs) in the central nervous system (CNS) in the human autoimmune disease multiple sclerosis (MS). In patients with secondary-progressive MS the presence of ectopic B cell aggregates has been linked to more severe clinical disease and cortical pathology. Here we demonstrate the formation of TLOs in myelin basic protein (MBP)-proteolipid protein (PLP) fusion protein MP4-induced experimental autoimmune encephalomyelitis (EAE) of C57BL/6 mice. TLOs were a characteristic feature of the chronic disease stage and most prevalent in the cerebellum, while also being present in the spinal cord and cerebrum, but absent in the brain stem. TLOs were characterized by B and T cell compartmentalization, the presence of high endothelial venules and germinal center formation. An association with TH17, but not TH1 cells was evident. In addition, results obtained from ELISPOT analysis of the antigen-specific T cell response suggest that TLOs facilitate determinant spreading of the MP4-specific T cell response to myelin oligodendrocyte glycoprotein (MOG). Our data add novel insights into the contribution of TLOs to disease progression in the context of CNS autoimmunity. We suggest that the blockade of TLO development is a therapeutic strategy that needs to be carefully considered in future studies.
Inflammation regulates the differentiation of CD4 T cells with cytolytic potential (P6088)
Authors: Brown Deborah , Workman Aspen , Vogel Alexander , Condon Shirley 
Published: January 2023

Abstract
Cytolytic CD4 T cells have been described during viral infections; however, the factors necessary for driving the cytolytic phenotype have not been identified. Our data indicate IL-2 regulates granzyme B (GrB) expression in CD4 T cells but IL-12 and IL-6 enhance GrB expression and promote perforin-mediated cytotoxicity in CD4 effectors. To further dissect the role of inflammation in CD4 CTL generation in vivo, we used an adoptive transfer system and infection with highly pathogenic influenza virus PR8, compared to x31 with lower pathogenicity. At relatively low doses (1000 EID50), Ova specific CD4 cells expressing GrB differentiated in PR8/Ova infected mice but not x31/Ova infected mice. This pattern was observed at 10-fold higher dose of x31/Ova compared to PR8/Ova. In addition, CD4 T cell influx to the lung and IFN-γ expression was increased in mice infected with PR8/Ova compared to x31/Ova. Importantly, these differences were seen in the endogenous, polyclonal population of CD4 T cells. Inflammatory mediators including IL-6, CXCL-9, CXCL-10 and MIP1β were 3-5 fold higher in PR8/Ova infected lungs at day 4 post infection compared to x31/Ova infected lungs. Whether these inflammatory cytokines are responsible for inducing CD4 CTL is being investigated. These data suggest that CD4 CTL are induced in vivo by strong inflammatory responses and may provide an extra layer of protection against highly pathogenic IAV infection.
Dynamic regulation correlates with variable inhibitory function of CD244 on antiviral CD8 T cells (P6117)
Authors: Pacheco Yovana , McLean Anna , Rohrbach Janine , Porichis Filippos , Kaufmann Daniel , Kavanagh Daniel 
Published: January 2023

Abstract
Antigen-experienced T cells express a wide variety of costimulatory molecules, some of which can contribute to activating, inhibitory, or exhaustion pathways depending on context. In particular 2B4 (CD244/ SLAMf4) has been shown to be capable of either inhibitory or enhancing effects upon engagement of its ligand CD48 (SLAMf2). We examined the phenotypes of CD8 T cell lines and clones derived from HIV-infected or -uninfected subjects, and specific for HIV or for respiratory syncytial virus. Antiviral CTL expressed varying levels of PD-1, 2B4, and TIM-3. Cognate peptide induced rapid downregulation of both 2B4 and TIM-3, but not PD-1 on cultured CD8 T cells. 2B4 downregulation also occurred directly ex vivo and required simultaneous signaling via both TCR and 2B4, as downregulation was not observed in the absence of antigen or in the presence of CD48-blocking antibody. 2B4 downregulation was not induced by PMA or prevented by PI3-kinase inhibition, implicating a TCR-proximal signaling mechanism. Downregulation occurred rapidly upon stimulation with a low dose of peptide, concomitant with internalization of CD3 and CD8. The degree of 2B4 downregulation varied among CD8 T cells of different specificities and HLA restrictions, and the degree of downregulation correlated with the respective enhancement or suppression of antiviral IFNγ production in response to CD48 blockade. Our results indicate a role for 2B4 downregulation in modulation of antiviral CD8 T cell responses.