bluuebunny/crossref_metadata_2025_split huggingface dataset:
B6.g7 mice reconstituted with BDC2.5NOD stem cells are protected against autoimmune diabetes. (P1392)
Authors: Rajasekaran Narendiran , Wang Nan , Hang Yan , Macaubas Claudia , Rinderknecht Cornelia , Beilhack Georg , Shizuru Judith , Mellins Elizabeth 
Published: January 2023

Abstract
In BDC2.5NOD mice, a spontaneous model of Type 1 diabetes, CD4+T cells express a transgene-encoded T-cell receptor with reactivity against a pancreatic antigen. This leads to a massive infiltration and destruction of the pancreatic islets and subsequent diabetes. When we reconstituted lethally irradiated lymphocyte-deficient B6.g7 (I-Ag7+) Rag-/- mice with BDC2.5NOD (ckit+Lin-Sca-1hi) hematopoietic stem and progenitor cells (HSPC), the recipients exhibited hyperglycemia and succumbed to diabetes. Surprisingly, lymphocyte-sufficient B6.g7 mice reconstituted with BDC2.5NOD HSPCs were protected from diabetes. In this study we investigated the factors responsible for attenuation of diabetes in the B6.g7 recipients. Analysis of chimerism in the B6.g7 recipients showed that although B cells and myeloid cells were 98% donor-derived, the CD4+ T cell compartment contained ~50% host-derived cells. These host-derived CD4+ T cells were enriched for conventional Tregs (CD25+Foxp3+) and also for host-derived CD25-Foxp3- CD4+ T cells that express markers of suppressive function, CD73, FR4 and CD39. Though negative selection did not eliminate donor-derived CD4+ T cells in the B6.g7 recipients, these cells were functionally suppressed. Thus host-derived CD4+ T cells that emerge in mice following myeloablation exhibit a regulatory phenoytpe and likely attenuate autoimmune diabetes. These cells may provide new therapeutic strategies to suppress autoimmunity.
Differential role of Bcl-2 in CD8+ and CD4+ T cell homeostasis (P1336)
Authors: Hildeman David , Raynor Jana , Tripathi Pulak , Sholl Allyson , Chougnet Claire 
Published: January 2023

Abstract
Regulated induction of apoptosis is critical for normal functioning of the adaptive immune system. Both TCR/MHC interactions and by soluble cytokines maintain T cells via their ability to modulate Bcl-2 family members, although the dependence of CD4+ and CD8+ T cells upon individual anti-apoptotic molecules remains unclear. Here, we used mice with a conditional allele of Bcl-2 crossed with dLck-Cre mice to assess the role of Bcl-2 in CD4+ and CD8+ T cell survival. We found that dlck-Cre-driven deletion of Bcl-2 resulted in a massive loss of naïve CD8+ T cells, but only a modest loss of naïve CD4+ T cells. We next tested if Bcl-2 was important for effector CD4+ T cell survival after infection with lymphocytic choriomeningitis virus (LCMV). Interestingly, the majority of LCMV-specific CD4+ T cells did not require Bcl-2 to survive contraction of the response. Further, treatment of mice with ABT-737, a Bcl-2, Bcl-xL, and Bcl-w inhibitor, only modestly increased the loss of effector CD4+ T cells, suggesting that neither Bcl-2, Bcl-xL, nor Bcl-w are required for survival of effector CD4+ T cells. Instead, we find that the Bcl-2 family member, A1 is increased in effector CD4+ T cells and we are currently investigating the role of A1 in the survival of naïve and activated CD4+ T cells. As the pro-apoptotic Bcl-2 family member Bim is critical to control naïve and effector CD4+ T cells, A1 may be an important Bim antagonist that promotes effector CD4+ T cell responses.
Host response in a vector transmission model of monocytotropic ehrlichiosis (P3353)
Authors: Saito Tais , Walker David 
Published: January 2023

Abstract
Ehrlichioses are emerging infectious diseases affecting veterinary and human hosts. Studies reveal potential pathogenic mechanisms of infection; however, not much is known about the role of the vector on disease establishment, as demonstrated by a needle-challenged vaccine but failed in field challenge by infected ticks. We have developed a tick transmission model for ehrlichial infection to study the vector-pathogen-host interaction. We infected mice by intravenous (IV) and intradermal (ID) routes and compared the subsequent infection with tick transmission of the newly described E. muris-like agent. We collected samples during the course of infection to evaluate bacterial levels in tissues and antibody production to characterize the differences induced by the route of infection. Our results showed bacterial levels and distribution patterns comparable between mice experimentally infected by the ID or IV route. Higher IgM antibody levels were observed in the early stage of infection in mice infected by tick transmission compared to ID and IV routes. Moreover, IgG antibody levels in tick-transmitted ehrlichiosis were higher than in ID inoculated mice and much higher than in IV infection, which showed only low levels during both early and later phases of infection. These initial results suggest important differences in host processing and presentation of ehrlichial antigen due to the route of infection, which will be further explored in our vaccine development research.
Identification of novel autoantibodies in systemic autoimmunity using 10,000-antigen proteome arrays (P4006)
Authors: Li Yajuan , Wang Ling , Zhou Jinchun , Mayeux Jessica , Lian Yun , Mohan Chandra , Li Quanzhen 
Published: January 2023

Abstract
Systemic erythematosus lupus (SLE) is an autoimmune disease characterized by presence of autoantibodies against a broad spectrum of self-antigens. In order to identify novel serum autoantibodies associated with SLE, we utilized a protoarray bearing 10,000 antigens to screen IgG and IgM autoAbs in sera of SLE patients. 446 IgG and 1218 IgM autoAbs were significantly increased in SLE compared with health controls (p&lt;0.05), in which 367 autAbs overlapped in IgG and IgM. Except the autoAbs previously identified in SLE patients, such as anti-DNA, Ro, La, etc., the protoarray also uncovered a group of novel autoAbs against a broad range of antigens. The increased expression of 16 novel autoantibodies in SLE, including IgG autoantibodies against APEX1, AURKA, BAG3, CSNK1G1, EIF2C1, HMGB1, IFIT5, MAPKAPK3, PADI4, PKRKA, RALGPS1, RGS3, SRP19, STIP1, UBE2S and VRK1 were further confirmed by ELISA. Gene expression analysis revealed that some of the self-antigens also significantly upregulated on transcription level. In addition to uncovering ~500 novel autoantibody specificities, the novel arrays also shed light on some of the pathogenic pathways leading to disease in systemic lupus. In conclusion, protoarray provide a powerful tool for identification of novel autoantibodies in system autoimmune diseases.
Antigen cross-presentation via Sec61 is indispensable for the development of lupus nephritis: a novel ‘self-organized criticality theory’ explaining the cause of SLE (P4014)
Authors: Tsumiyama Ken , Shiozawa Shunichi 
Published: January 2023

Abstract
Objective: In our ‘self-organized criticality theory’ of autoimmunity, CTL develops via antigen cross-presentation and causes lupus tissue injuries. Here we show an essential role of a translocon Sec61 for antigen cross-presentation and lupus nephritis. Methods: Bone marrow-derived dendritic cell (BMDC) from BALB/c mice was cultured with fluorescent-labeled OVA. Early endosome antigen 1 (EEA1), calnexin and Sec61 were detected by immunofluorescent staining. For in vivo study, BALB/c mice were repeatedly immunized with OVA to induce SLE. The splenic DC (spDC) was also cultured with fluorescent-labeled OVA to examine localization of engulfed antigen. Further, exotoxin A was co-immunized with OVA to inhibit Sec61 in vivo, and IFNγ-producing CD8+ T cell and proteinuria were examined. Results: In BMDC, OVA was co-localized with an endosomal marker EEA1, and subsequently separated from EEA1. OVA was not co-localized with an ER marker calnexin. Instead, OVA was co-localized with Sec61. The same result was obtained using spDC, suggesting that OVA could be exported from endosome to cytoplasm via Sec61. After repeated immunization with OVA, the amount of Sec61 localized in endosome was increased compared with control. Further, treatment with a Sec61 inhibitor exotoxin A inhibited the generation of CTL and lupus nephritis. Conclusion: Direct export of antigen from endosome to cytoplasm via Sec61 is essential for antigen cross-presentation and the development of lupus nephritis.
Plasmacytoid dendritic cells correlate with fibrosis in patients with systemic sclerosis and contribute to fibrosis in a murine model (P3133)
Authors: Kafaja Suzanne , Divekar Anagha , Valera Isela , Khanna Dinesh , Saggar Rajan , Furst Daniel , Singh Ram 
Published: January 2023

Abstract
Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis in multiple organs including skin and lung. Mechanisms that drive fibrosis in SSc remain unclear. Here, we report the role of dendritic cells (DC) in SSc, with a focus on plasmacytoid DC (pDC) subset. We found that whereas pDC were reduced in the blood of patients with SSc as compared to healthy controls, these cells accumulated in target organs such as skin and lungs. In the lung lavage fluid (BAL), pDC frequency correlated with CT scores of SSc-interstitial lung disease (ILD) and with a set of proteins involved in inflammation, leukocyte migration, and wound repair. Importantly, pDC were reduced in the BAL of SSc patients after treatment with imatinib mesylate that has been previously reported to improve, albeit modestly, skin score and lung function. To directly investigate the role of pDC in vivo, we depleted pDC in mice injected with bleomycin that induces SSc-like disease. pDC-depleted mice had reduced skin and lung disease compared to controls. Thus, we provide several lines of evidence in humans and mice supporting a role of pDC in SSc pathogenesis. Ongoing studies will investigate mechanisms that lead to increased migration of pDC from blood to target organs and/or their local expansion in humans and mice with SSc.
Neurokinin-1 receptor stimulated dendritic cell vaccines downregulate IL-10 and license host inflammatory dendritic cells to bias Th1 immunity (P4236)
Authors: Sumpter Tina , Janelsins Brian , Rojas-Canales Darling , Tkacheva Olga , Shufesky William , Morelli Adrian , Larregina Adriana 
Published: January 2023

Abstract
Substance-P (SP) is a neuropeptide that stimulates immunity by binding to the neurokinin-1 receptor (NK1R). As professional Ag presenting cells, dendritic cells (DC) are unique activators of naïve T cells. DC express functional NK1R, indicating that NK1R agonists may influence their T cell-stimulatory function. Relevant studies analyzing the relationship between neuropeptides and DC function are lacking. Here, we demonstrate that conditioning bone marrow derived DC (BMDC) with NK1R agonists leads to maturation and abrogation of IL-10 without inducing secretion of IL-12p70. Mechanistic analyses in vitro demonstrated that NK1R signaling blocks the CREB1/TORC2 pathway to inhibit the Il-10 promoter. Following subcutaneous administration as a cellular vaccine, Ag-loaded and NK1R-signaled BMDC homed to skin draining lymph nodes (sDLN), initiating inflammation and Th1 and Tc1 immunity in vivo. This type-1 immunity required IL-12p70 secreted in sDLN by host DC, including LN-resident, and recruited inflammatory DC. Utilizing CCR7KO BMDC and IL-12p35KO mice, we elucidated that endogenous DC subsets were activated in situ by NK1R-signaled-BMDC homing to sDLN. Moreover, reduced levels of IL-10 by NK1R signaled BMDC licensed IL-12 secretion by host DC and mediated type-1 immunity. Our data demonstrate that NK1R-signaling promotes immunostimulatory DC and provide a relevant insight into the mechanisms employed by neuromediators to regulate the outcome of cellular immune responses.
Development of genetically engineered TCR-transduced T cells for immunotherapy of chronic hepatitis B and C virus infections (P4222)
Authors: Park Su-Hyung , Park Heiyoung , Pan Warren , Rehermann Barbara 
Published: January 2023

Abstract
More than 350 million people worldwide are chronically infected with hepatitis B (HBV) or hepatitis C virus (HCV). While vigorous CD8 T cell responses are required to spontaneously clear acute HBV and HCV infections, these virus-specific T cells are typically diminished in number and functionally impaired in chronic HBV and HCV infections. Here, we set out to use adoptive therapy with TCR-engineered T cells to restore HBV and HCV-specific immune responses. Three HBV-specific and four HCV-specific TCRs with high avidity were cloned from patients and chimpanzees that cleared the respective infection, expressed in retroviral vectors and used to transduce lymphocytes of chimpanzees with chronic HBV and HCV infection. The transduced T cells gained a broad spectrum of antigen-specific effector functions (IFN-γ secretion, cytotoxicity and proliferation). They were expanded to high numbers via short-term anti-CD3 and IL-15/IL-21 stimulation, which resulted in a preservation of their differentiation status and a lower frequency of regulatory T cells than the conventional expansion method with anti-CD3 and IL-2. Collectively, we show that retroviral TCR gene transfer followed by short-term expansion in the presence of IL-15/IL-21 efficiently redirects the antigen specificity of resting primary T cells and generates a large number of functional effector T cells. These will be used in adoptive T cell therapy protocols to restore T cell function in chronic viral hepatitis.
Antigen-specific natural killer cell cytokine production in chronic hepatitis C infection (P6062)
Authors: Bowman Kathryn , Holz Lauren , Rehermann Barbara 
Published: January 2023

Abstract
Chronic hepatitis C virus (HCV) infection results in an inflammatory liver disease leading to fibrosis and cirrhosis. The progression of liver disease is thought to be immune-mediated because HCV itself is non-cytopathic. Given that HCV-specific T cells are diminished in number and functionally exhausted in chronic HCV infection, it remains unclear which cell population drives disease pathogenesis. Here, we investigated the function of natural killer (NK) cells, the major innate immune cell population in the liver. PBMCs from 15 patients with chronic HCV infection were stimulated for 8 hours in a whole blood activation assay with pools of overlapping 18-mer peptides comprising HCV structural (E1, E2) and nonstructural (NS3) proteins. Cytokine production by NK cells and T cells was assessed by multicolor flow cytometry. The frequency of IFN-γ+ NK cells was 5-10 fold greater than the frequency of IFN-γ+ T cells. A minority of IFN-γ+ NK cells co-produced TNF-α. NK cell responses to HCV peptides varied between subjects, but did not correlate with T cell responses or viremia. This study demonstrates that NK cells are activated in an antigen-specific manner in chronic HCV infection and respond to both structural and nonstructural HCV proteins. The mechanism of antigen-specific NK cell activation is currently under investigation.
An analysis of expression of lupus-associated miRNAs in splenic lymphocytes from male and female NZB/WF1 mice (P4074)
Authors: Dai Rujuan , McReynolds Savannah , Heid Bettina , Ahmed S.Ansar 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) is a prototype, female predominant autoimmune disease. The functional importance and pathogenic contribution of microRNAs (miRNAs) to lupus are now highly recognized. We reported recently a common set of lupus-associated miRNAs including miR-182-96-183 cluster, miR-155, miR-31, and miR-127 in three spontaneous murine lupus models including NZB/WF1. To understand whether miRNAs are differentially expressed in males versus females, and thereby contribute to increased susceptibility to lupus in female NZB/WF1, we analyzed the expression of above miRNAs in splenic lymphocytes from male and female NZB/WF1 mice at different disease stages. Our data indicated that there is no significant difference in the expression of above lupus-associated miRNAs between female and male NZB/WF1 at 17-18wk old, an age before the onset of disease. However, by 23 weeks of age (when NZB/WF1 begin to develop mild lupus disease) the expression levels of miR-182, miR-183, miR-127, and miR-31 were significantly up-regulated in female NZB/WF1 mice compared to age-matched male controls. The expression levels of these miRNAs were further increased in 30 wk old females, following the progression of lupus disease. There was also a slight increasing trend of the above miRNAs in 30 wk old male NZB/WF1 mice when compared to 23 wk old male mice, however it was not significant. Taken together, our study further suggested the association of these miRNAs with lupus development.
Contribution of novel microRNAs in the pathogenesis of rheumatoid arthritis (P4078)
Authors: Singh Ram , Hahn Bevra , Hossain Awlad 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a systemic inflammatory autoimmune disease with progressive joint destruction, lifelong disability, functional deterioration, and high mortality. The major effector cells in RA are T cells and synovial fibroblasts. Defects in T cells and synovial fibroblasts lead to activation of proinflammatory cytokines, which induce Th1 and Th17 cells and chronic inflammation. Recent studies provide further evidence of dysregulation of candidate microRNAs in inflammation and autoimmunity. In our preliminary studies we have found dysregulated candidate microRNA (miRNA21, miR25 miR95, miR124a, miR146a and miR186) expression in RA patients compared to healthy controls. Furthermore proinflammatory cytokines such as IL-6, IL-12, IL-17 and IL-21 are significantly increased in RA patients compared to healthy controls, and expression of these cytokines positively correlates with the expression of candidate microRNAs such as miR21 and negatively correlates with the expression of miR146a. Furthermore we found that miR21 and miR124a positively correlate, and miR146a negatively correlates, with DAS28 score in RA patients. Based on these preliminary data, we hypothesize that dysregulated microRNAs play an important role in RA pathogenesis. This study has translational potential and may identify novel therapeutic targets in patients with RA.
TNF receptor superfamily members drive thymic Treg development (P4430)
Authors: Mahmud Shawn , Manlove Luke , Schmitz Heather , Hogquist Kristin , Farrar Michael 
Published: January 2023

Abstract
CD4+FOXP3+ regulatory T cells (Tregs) express several TNF receptor superfamily (TNFRSF) members, but what role these molecules play in thymic Treg development is unclear. We found that CD25+FOXP3- Treg progenitors and CD25+FOXP3+ thymic Tregs highly overexpress OX40, GITR, and TNFR2 relative to conventional CD4SP thymocytes. Utilizing a novel Nur77GFPBac reporter mouse, which reports TCR signal strength via GFP, we found that OX40, GITR, TNFR2 and CD27 expression by Treg progenitors positively correlates with TCR signal strength. Importantly, the ligands for these receptors are expressed by APCs in the thymic medulla. Antibody-blockade of TNFRSF members in thymic organ cultures reduced expression of CD25 and FOXP3 by Tregs. Conversely, stimulation of sorted Treg progenitors with TNFSF agonists enhanced Treg differentiation ~3-4 fold and elevated CD25 and Bcl-2 expression. In vivo, GITR- or OX40-deficiency reduced Treg development by ~50%. Finally, dominant negative retroviral bone marrow chimeras designed to knock out up to three TNFRSF members simultaneously showed a dose dependent (up to ~6 fold) reduction in Treg development indicating that these receptors function redundantly. Our data support a model in which TNFRSF expression driven by TCR signaling enhances IL2 sensitivity and promotes survival in Treg progenitors to enhance thymic Treg differentiation.
A clinical, epidemiological and immunological characterization of a dengue cohort in a metropolitan city in Malaysia (P3027)
Authors: Sekaran Shamala Devi , Rathakrishnan Anusyah , Azliyati Azizan , Sathar Jameela , Sanchez-Anguiano Aurora , Kumari Santha 
Published: January 2023

Abstract
Dengue has neither licensed drugs nor vaccines and Its immunopathogenesis is elusive. The disease is widespread and imposes healthcare, economic and social burden on endemic countries like Malaysia. This study aims to understand and identify clinical and immunological markers of disease. Dengue suspected patients were recruited in 2 hospitals where 77% of the study population was categorized as dengue with warning signs. Diagnosis based solely on clinical symptoms is insufficient. Clinically, platelet counts of dengue patients were low and together with increased vWF and prolonged clotting time, retain thrombocytopenia in dengue as an excellent marker for disease progression but not as a differentiator for patients with and without warning signs. Liver enzymes, ALT and AST, were good indicators for dengue disease progression. Immunogenetically, HLA alleles A*24 and B*57 and A*03 were associated with Chinese, Indians and Malay dengue patients, respectively. Three non-structural proteins were found to be immunodominant via the IFN-γ T cell ELISpot assay while cytokines, TNF-β, MIF, SDF-1α and HGF had differential levels in patients with and without warning signs. Although we did observe any discriminative clinical markers, we detected differential levels of 4 cytokines between the groups of patients, which may, with further studies, serve as predictive markers for dengue patients developing warning signs.
MicroRNA miR-146a is a critical suppressor of arthritis in the mouse model of Lyme Disease (P1275)
Authors: Lochhead Robert , O'Connell Ryan , Ma Ying , Zachary James , Weis John , Weis Janis 
Published: January 2023

Abstract
Lyme Disease is caused by infection with the spirochete Borrelia burgdorferi, and is the most common vector-borne disease in the United States. Often, infection leads to acute arthritis in humans. Our lab and others have exploited the mouse model system to elucidate the mechanical mechanisms of Lyme Disease development. Because miRNAs have been shown to be important regulators of inflammatory and immune responses, we performed a microarray screen for expression of miRNAs in joint tissue from three infected mouse strains. This screen identified upregulation of miR-146a, a key negative regulator of NF-κB signaling. Others and we have previously demonstrated that Toll-like receptor 2/MyD88 dependent activation of NF-κB is a major regulator of immune response in mice and humans, but determining its role in arthritis development has remained elusive. We show that miR-146a plays a critical nonreduntant role in modulating Borrelia-induced NF- κB response to infection, and mice lacking this miRNA develop more severe arthritis without compromising immune response. Furthermore, miR-146a-/- mice have elevated transcript levels of NF-κB-regulated cytokines and chemokines in joint tissue, as well as increased myeloid cell infiltration, linking NF-κB dysregulation to arthritis development. Together, these data show that miR-146a is required for suppression of Lyme Arthritis, and provides a much-needed model for studying the role of NF-κB activation in arthritis development.