bluuebunny/crossref_metadata_2025_split huggingface dataset:
Liposomal vaccine formulations incorporating CpG enhance multiple parameters within the lymphatic system that improve immunity (VAC8P.999)
Authors: Neeland Melanie , Meeusen Els , de Veer Michael 
Published: January 2023

Abstract
Liposomal vaccine formulations incorporating stimulants that target innate immune receptors have been shown to significantly increase vaccine induced immunity. Following vaccination, innate cell populations carry antigen from the injection site, via the afferent lymphatic vessels, into the local lymph node. Antigen-specific and activated lymphocytes then leave the lymph node via the efferent lymphatics to perform their effector function. Using unique in vivo models of ovine lymphatic cannulation, this study investigates the immune response occurring in afferent and efferent lymph after vaccination of liposomal delivery systems incorporating the innate stimulator CpG. We show that the addition of CpG to liposomal vaccine formulations enhances DC associated antigen transport and uniquely increases MHC II expression on antigen carrying monocytes. RNA sequencing experiments revealed that 72h after injection of CpG, signatures associated with inflammation, interferon signaling and Th17 cell differentiation are significantly increased in cells migrating to the local lymph node. CpG also promotes the persistence of antigen specific antibodies and significantly increases IFNγ production by CD8 T cells. The results of this study demonstrate the real time in vivo kinetics of the peripheral contribution to liposome based delivery systems and add to our understanding of the immunogenic function of poly(I:C) and CpG when employed as adjuvants within liposomal vaccine formulations
Intranasal vaccination with live attenuated influenza vaccine increases diversity of the human nasal microbiome (VAC8P.1001)
Authors: Lambert Nathaniel , Salk Hannah , Kennedy Rick , Chia Nicholas , Grill Diane , Jacobson Robert , Poland Gregory 
Published: January 2023

Abstract
Pre and post-licensure studies have supported the safety of the intranasally administered live, attenuated influenza vaccine (LAIV). The objective of this study was to measure changes to the nasal microbiome in subjects who received LAIV. We hypothesized that administration would result in only a transient alteration of the nasal microbiome. To test this hypothesis, we swabbed the anterior nares just before vaccination (Day 0), and on Day 7 and Day 28. We performed next-generation sequencing on amplified 16s rRNA genes. There was a significant increase in alpha diversity (Observed, CHAO, and ACE) from Days 0 to 7 (p = 0.02, 0.00, 0.01; respectively) and Days 0 to 28 (p = 0.05, 0.03, 0.05; respectively). When measuring differences in specific observed taxanomic units (OTUs), we identified a significant increase in Flavobacteriaceae and Enterobacteriaceae after vaccination. These data suggest that LAIV may increase the ability of these less abundant OTUs to establish a community niche and, therefore, could be clinically relevant. We have begun to investigate the second objective of this study, which is to correlate microbiome composition with mucosal immunity to influenza virus. To our knowledge, this is the first study to investigate LAIV-induced alterations to the human microbiome. These data may lead to a better understanding of the influences of the host microbiome on the response to vaccine, and perhaps infection.
Enhanced anti-retroviral cytotoxic T cell response by PD-1 blockade (VIR4P.1015)
Authors: Li Wen , Green William 
Published: January 2023

Abstract
LP-BM5 retrovirus induces a complex disease featuring an acquired immunodeficiency syndrome termed murine AIDS (MAIDS) in susceptible strains of mice, such as C57BL/6 (B6). Our lab has previously shown that a cytotoxicT lymphocyte (CTL) response to MAIDS-associated epitopes that are shared by MAIDS-associated terminal B-cell lymphomas can be detected in uninfected, MAIDS-associated tumor cell primed B6 mice. We further confirmed that B6 mice infected with LP-BM5 retrovirus can generate equally robust anti-MAIDS tumor CTLs compared to the B6 mice primed with tumor cells. To determine if the anti-tumor CD8 T-cell responses can be enhanced by interrupting the down-regulatory PD-1 pathway specifically in the CD8 T-cell compartment, thus to treat MAIDS-associated B-cell lymphomas, we determined whether adoptive transfer of PD-1KO CD8 T cells with or without total CD4 T cells, was able to protect Rag-1KO mice from B6-1710 tumor challenge. Rag-1KO recipients that received B6.PD-1KO.CD8 T cells showed delayed tumor growth following B6-1710 tumor challenge compared to Rag-1KO mice that received equal numbers of w.t B6.CD8 T cells. This enhanced anti-tumor activity was similar to what was observed in mice that received combined PD-1 blockade in CD8 T cell compartment and depletion of CD4 T regulatory cells. Our data indicate that anti-tumor CD8 T cell response can be increased by interrupting the down-regulatory, CD8 T-cell mediated PD-1 pathway.
CNS stromal cells modulate antiviral CD8 T cell immunity (VIR4P.1009)
Authors: Swanson Phillip , McGavern Dorian 
Published: January 2023

Abstract
Lymphocytes are essential for control of neurotropic viruses such as HSV, HIV, and JC virus. Understanding the mechanisms involved in the regulation of these cells is essential for the development of therapeutic interventions to treat central nervous system (CNS) infections. Using two-photon microscopy, we observed that CD8 T cells responding to a viral infection interact with meningeal stromal cells (MSCs), a population of specialized fibroblasts that comprise a dense, collagenous fiber network within the lining of the brain. Little is known about the immunological functions of MSCs. To gain novel insights into these cells, we conducted genomic analyses of purified MSCs at different time points following CNS infection with lymphocytic choriomeningitis virus (LCMV). Our analyses revealed that MSCs responded to virus infection through induction of a robust immunological program that included genes associated with antigen presentation, chemotaxis, immune regulation, among others. We also observed in vivo that MSCs were able to present viral-peptide MHC I complexes, which corroborated our intravital imaging studies showing direct, stable interactions between MSCs and anti-viral T cells. MSCs possess the capacity to regulate these interactions through expression of multiple immune regulators such as PD-L1, galectin-9, and PVR. Our data demonstrate that MSCs are a novel CNS-resident antigen presenting cell with the ability to modulate antiviral T cells following infection.
Anti-viral effects of CGRP in mixed glial cells infected with LP-BM5 virus (VIR5P.1022)
Authors: Grlickova-Duzevik Eliza , Malon Jennifer , Cao Ling 
Published: January 2023

Abstract
LP-BM5 retrovirus infection is an established murine model used to study HIV infection in humans due to that susceptible strains of mice (including C57BL/6(B6) mice) develop immunodeficiency syndrome (termed as MAIDS) following LP-BM5 infection. Previously, we observed that calcitonin gene related protein (CGRP), a neuropeptide produced primarily by sensory neurons, reduced LP-BM5 viral loads in infected mixed glial cells (primarily astrocytes and microglia). The mechanism through which CGRP decreases the viral loads is still not known. To elucidate the role of CGRP in glial LP-BM5 infection, we first determined that CGRP did not induce an increased cell death post-infection or directly decrease the viral infectivity. Then, we investigated whether CGRP could affect viral infectivity by enhancing the glycosylation of the cell receptor for viral attachment, murine cationic amino acid transporter 1 (mCAT-1). Our data indicate that blocking glycosylation by Tunicamycin did not affect CGRP’s anti-viral effects. Further, we tested the involvement of p38 MAPK pathway in CGRP-induced inhibitory effects. Preliminary experiment using a p38 inhibitor, SB203580, suggests a p38 mediated mechanism. Future studies will further elucidate the role of p38 and examine the role of other CGRP downstream pathways on LPBM5 infection.
Macrophage depletion ameliorates nephritis induced by pathogenic antibodies (BA12P.111)
Authors: Chalmers Samantha , Chitu Violeta , Stanley E. Richard , Putterman Chaim 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by the production of autoantibodies and inflammatory damage in target organs. Kidney involvement affects up to 60% of lupus patients, and is responsible for significant morbidity and mortality. Previous studies using methods to reduce macrophage number have reached conflicting conclusions regarding the role of macrophages in lupus nephritis (LN). Moreover, “off target” effects occur in mice congenitally deficient in macrophages. In this study, we investigated the role of macrophages in mice receiving anti-glomerular antibodies, or nephrotoxic serum (NTS), an experimental model which closely mimics the immune complex mediated disease seen in LN. GW2580, an oral kinase inhibitor monospecific for the CSF-1 receptor, was used as a novel and highly selective method for macrophage depletion. We found that GW2580-treated NTS challenged mice did not develop the significant increases in levels of proteinuria, serum creatinine or serum urea, seen in control treated mice. Further, GW2580-treated mice were protected from the robust kidney expression of inflammatory cytokines associated with LN seen in control treated mice, including RANTES, IP-10, VCAM1, MCP-1 and IL-6. IBA-1 staining confirmed profound depletion of macrophages in glomeruli of GW2580-treated mice. Our results support an important role of macrophages in LN, and suggest targeting this cell type as a promising approach to the treatment of LN.
Pathogenesis of cell death in lupus bone marrow (BA12P.112)
Authors: Zhuang Haoyang , Han Shuhong , Xu Yuan , Li Yi , Yang Lijun , Reeves Westley 
Published: January 2023

Abstract
Pristane-treated mice develop lupus mediated by TLR7-driven production of IFNα (autoantibodies, nephritis) and TNFα (anemia). Anemia is IFN-independent, is caused by TNFα produced by bone marrow (BM) neutrophils, and is associated with a striking accumulation of dead BM niche cells and erythroid precursors in the BM. TNFα and dead cells also were found in BM from anemic SLE patients. The dead cells may drive TNFα/IFNα production by releasing TLR7 ligands. Fas-deficient (B6/lpr) mice were completely refractory to induction of pristane-lupus implicating cell death in its pathogenesis. Interestingly, accumulation of dead cells was abolished in BM from B6 TNFα-/- mice, suggesting that they were killed by TNFα. Signaling through both Fas and the TNF receptor I can activate caspase-3 and processing of the BCL-2 family member BID, which is involved in a mitochondrial amplification loop for Fas-mediated apoptosis. However, pristane injection in BID-/- mice induced TNFα and IFNα production as well as autoantibody production comparable to wild type B6 mice. We conclude that the requirement for Fas signaling in pristane-lupus involves the death of “Type I” cells, which undergo Fas-mediated apoptosis without a requirement for truncated BID and its crosstalk with the mitochondrial cell death pathway.
Autoreactive B cell tetramer analysis reveals a type I interferon-mediated marginal zone precursor B cell defect in autoimmune BXD2 mice (BA12P.102)
Authors: Hamilton Jennie , Li Jun , Wu Qi , Yang PingAr , Luo Bao , Bradley John , Taylor Justin , Randall Troy , Mountz John , Hsu Hui-Chen 
Published: January 2023

Abstract
In human systemic lupus erythematosus and rheumatoid arthritis, development of self-reactive B cells has been associated with defective B-cell checkpoints prior to somatic hypermutation and class-switch recombination in germinal centers (GCs). To identify mechanisms leading to checkpoint defects in the periphery, sera from BXD2 autoimmune mice were screened on a PEPperPRINT Microarray. Highly specific autoepitopes were narrowed down via Discotope epitope analysis and high-throughput ELISA and ELISPOT analyses. Three peptides La, snRNP, and fibromodulin-citrulline (fib-cit) were used to make tetramers (tet) for characterization of B cell subsets in the spleen. Analysis of fib-cit tet+ B cells revealed an age-dependent 5 fold increase in tet+ B cells, especially within the marginal zone-precursor (MZ-P) (10.6% vs 4.85%) and GC (7.3% vs 0%) subsets, in BXD2 vs B6 mice. While the total frequency of La and snRNP tet+ B cells was not different in BXD2 vs B6, there was also an increase in the MZ-P (7.66% vs 1.51%) phenotype. Sorted tet+ B cells exhibited a &gt;100-fold enhanced ability to secrete specific autoAb compared to tet- B cells. Additionally, there was significantly higher CD69 expression (75.5% vs 5.71%), a type I interferon (IFN)-induced protein in MZ-P B cells in BXD2 vs B6 mice. Together, these results establish a general method for characterization of autoAg specific B cells and suggest a checkpoint defect at the MZ-P stage as a result of type I IFN stimulation.
Redox regulation of a new autoimmune mouse model, glucose-6-phosphate isomerase peptide induced arthritis in mice (BA12P.108)
Authors: Yang Min , Holmdahl Rikard 
Published: January 2023

Abstract
Human glucose-6-phosphate isomerase (hG6PI) peptide-induced arthritis is established by immunizing C57Bl/6N.Q (B6N.Q) mice with single hGPI325-339 peptide emulsified in complete freund’s adjuvant. Immunized B6N.Q mice start to develop arthritis on day 10-12, reach to the peak value of severity on day 14-20 and then recover during the next 7-10 days. This peptide-induced arthritis mouse model represents an acute arthritic disease progression. By taking advantage of μMT mice (B cell deficient) and TCRβ-/- (T cell deficient) mice, we have demonstrated that this peptide-induced arthritis is T cell- and B cell- dependent. In addition, by using Ncf1 mutant mice, which are lack of oxidative burst, we have found that reduced reactive oxygen specifics aggravate and prolong the peptide-induced arthritis. Phenotypic analysis results show that increased frequency and absolute numbers of Th17 cells, accompanied by significant increased CD11b+cells, are observed in Ncf1 mutant mice comparing with wild-type mice; moreover, IL-17 levels were dramatically increased in isolated splenic cells or lymph nodes cells from immunized Ncf1 mutant mice when restimulated with hGPI325-339 peptide. Moreover, in vitro experiments suggest that APCs from Ncf1 mutant mice show higher capability to present hGPI325-339 peptide to stimulate hGPI325-339 peptide specific T cell hybridoma, which suggest that enhanced antigen-presentation capability may aggravate the arthritic disease in Ncf1 mutant mice.
TNF receptor superfamily costimulation couples TCR signal strength to thymic Treg differentiation (BA13P.128)
Authors: Farrar Michael , Mahmud Shawn , Manlove Luke , Schmitz Heather , Xing Yan , Wang Yanyan , Owen David , Schenkel Jason , Boomer Jonathan , Green Jonathan , Yagita Hideo , Chi Hongbo , Hogquist Kristin 
Published: January 2023

Abstract
Regulatory T cells express several TNF receptor superfamily (TNFRSF) members, but their role in thymic Treg development is undefined. We demonstrate that Treg progenitors express high levels of the TNFRSF members GITR, OX40, and TNFR2. Expression of these receptors requires TAK1 and CD28 and correlates directly with TCR signal strength. Neutralizing TNFSF ligands markedly reduced Treg development. Conversely, TNFRSF agonists enhanced Treg differentiation by augmenting IL2R/STAT5 responsiveness. Importantly, GITR-L led to a dose-dependent enrichment of lower-affinity cells within the Treg repertoire. In vivo, GITR- or OX40-deficiency modestly attenuated Treg development; in contrast, combined inhibition of GITR, OX40 and TNFR2 abrogated Treg development. Thus TNFRSF expression on Treg progenitors translates strong TCR signals into molecular parameters that specifically promote Treg differentiation and shape the Treg repertoire.
Regulation of organ-specific inflammation by CXCR5 during systemic autoimmunity (BA13P.131)
Authors: Sharma Rahul , Abaya Christian , Sharma Poonam , Stremska Marta , Jose Sheethal 
Published: January 2023

Abstract
Mice and humans deficient in the Foxp3+ T regulatory cells (Tregs) develop systemic autoimmunity affecting multiple organs and die prematurely. Mice deficient for interleukin (IL)-2 (IL2KO) have reduced Tregs and develop inflammation mainly in liver, colon, pancreas and salivary glands. We found that the CD4+ T-cells in the secondary lymphoid organs IL2KO mice highly express CXCR5, a chemokine receptor and a marker for T-follicular helper (TfH) cells. We hypothesized that CXCR5 expression may regulate inflammation by regulating T-cell recruitment and organ-specific autoantibody production. We found that CXCR5+CD4+ T-cells were highly enriched in these inflamed organs of IL2KO mice. Rendering the IL2KO mice deficient for CXCR5 led to prolonged lifespan compared to CXCR5-sufficient IL2KO mice and protection against inflammation especially in liver and colon and partial protection against inflammation in pancreas and salivary glands. This was accompanied with a reduction of activated B and T cells in the secondary lymphoid organs as well as reduced production of inflammatory cytokines by the T-cells. However, no significant correlation was observed between CXCR5 expression and antibodies to liver and pancreatic auto-antigens. The data suggests that CXCR5 expression regulates organ-specific inflammation during Treg-deficiency -induced systemic autoimmunity, through regulation of T-cell trafficking, with an undetectable contribution to autoantibody production.
Novel function of ECM1 on inhibiting Th17 differentiation and infiltration in EAE development (BA13P.116)
Authors: Sun Bing , Su Pan , Zhou Haiyan , Zhang Yaguang 
Published: January 2023

Abstract
Our previous work has demonstrated that Extra-cellular Matrix Protein 1 (ECM1) is a specific secreted protein by Th2 cells (Nature Immunology, 2011). In this study, we demonstrated that ECM1 protein is capable of inhibiting Th17-type cell differentiation during EAE initiation and then prevents EAE development. The mechanisms study showed that ECM1 suppresses TGF-b maturation and in turn to inhibit Th17-type cell differentiation. It is also observed that ECM1 can inhibit Th17 cell infiltration into neuron central system through targeting MMP9. Our results indicate that ECM1 has a novel function to suppress Th17 cell differentiation and infiltration in EAE mice model.
Disruption of maternal tolerance during pregnancy leads to Treg repopulation of the antigenic UPI (BA13P.123)
Authors: Collier Ai-ris , Snead Andrea , Cook Heather , Yesayan Maria , Kahn Daniel 
Published: January 2023

Abstract
Background Regulatory T cells (Treg) are essential for normal pregnancy outcome. In vivo, Treg depletion during pregnancy leads to autoimmunity that results in total pregnancy loss. The paucity of Treg and effector cells in normal pregnancy uteroplacental interfaces (UPI) renders the mechanism unclear. Methods Low-dose (0.1ug) diphtheria toxin was given on GD11 to female C57-Bl6 FoxP3-DTR under a syngeneic pregnancy. On day 17, pup weight and gender was determined and UPI were removed and subject to FACS and confocal immunofluorescence. Results Subtle Treg depletion during pregnancy resulted in effector T cell influx preferentially to UPIs of antigenic (male) fetuses resulting in a reduction of fetal weights. Surprisingly, we also observed an increase of Treg numbers in the antigenic UPIs. Preliminary results indicate a local interaction between antigen-presenting cells, T cells and Treg in the previously depleted UPI. Conclusions Systemic interruption of Treg tolerance during pregnancy allows for maternal attack on antigenic fetuses. In the unperturbed UPI, the absence of Treg suggests that maternal tolerance is mediated elsewhere. In disrupted maternal tolerance, the repopulation of Treg to the UPI provides insight into Treg cellular interactions that may contribute to the re-establishment of tolerance under physiologic conditions.
The type 1 diabetes resistance locus B10 Idd9.3 impairs B cell development and implicates microRNA-34a as a regulator of T1D pathogenesis (BA14P.200)
Authors: Waldner Hanspeter , Berry Gregory , Budgeon Lynn , Cooper Timothy , Christensen Neil 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune disease resulting from the T cell-mediated destruction of insulin-producing pancreatic beta cells. Multiple insulin-dependent diabetes (Idd) loci confer T1D susceptibility or resistance in both humans and nonobese diabetic (NOD) mice, but the genes and mechanisms by which they control disease development are largely unknown. NOD.B10 Idd9.3 mice contain the Idd9.3 locus from T1D-resistant C57BL/10 mice that encodes 19 genes, including microRNA (miR)-34a, and confers partial T1D protection. B cells have been shown to play a critical role in priming autoantigen-specific CD4+ T cells in NOD mice. We show that early B cell development is impaired in NOD.B10 Idd9.3 mice, resulting in significant reduction of splenic B cells compared to NOD mice. Molecular analysis revealed that miR-34a expression was significantly elevated in B cell progenitors and marginal zone B cells of NOD.B10 Idd9.3 mice compared to NOD mice. Furthermore, miR-34a expression inversely correlated with levels of Foxp1, an essential regulator of B cell lymphopoiesis, which is directly repressed by miR-34a. Importantly, peripheral B cell paucity in NOD.B10 Idd9.3 mice was associated with impaired proliferation of islet-antigen-specific CD4+ T cells in response to endogenous autoantigen in pancreatic lymph nodes. These data suggest that miR-34a negatively regulates B cell development in NOD.B10 Idd9.3 mice and thereby may contribute to their T1D protection.
Induced pancreatic inflammation is controlled in mice by extrapancreatic insulin expression and CD8 T cells limiting autoimmune attack (BA14P.209)
Authors: Girtman Megan , Pavelko Kevin , Pease Larry 
Published: January 2023

Abstract
Type 1 diabetes can be induced by virus expressing SIINFEKL in Rat Isulin Promoter (RIP)-OVA transgenic mice harboring adoptively transferred naïve antigen-specific OT1 T cells. By titrating the number of transferred OT-1 cells, disease outcome can be manipulated. Mice receiving high numbers of OT-1 cells develop autoimmunity and rapidly succumb to disease. Mice receiving limiting numbers develop hyperglycemia transiently and then over 10 days recover glucose homeostasis. Recovered mice exhibit restored normal resting blood sugar levels and are able to regulate glucose during a stress test. Histological examination of the pancreas showed extensive pancreatic destruction with no or very few remaining insulin-positive islets. Using RIP-OVA mice expressing luciferase under the mouse insulin promoter, pancreatic insulin promoter activity was extinguished, while liver expression increased in recovered animals, suggesting an ectopic source of insulin can support glucose homeostasis in mice with extensively damaged islets. Pancreatic infiltrates during peak inflammation contained host-derived CD8 T cells in addition to the infiltrating OT-1 cells. These host CD8 T cells were absent in CD8 deficient RIP-OVA mice, and diabetes occurred with fewer transferred OT-1 cells and was quickly fatal. Reconstitution of the CD8 compartment protected these animals. These findings indicate the importance of host CD8 T cells in limiting autoreactive T cell-mediated destruction of islets.