bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD11c-expressing B cells from aged and autoimmune mice localize at T cell B cell border in spleen and present antigen more efficiently than Follicular B cells (BA3P.104)
Authors: Rubtsov Anatoly , Rubtsova Kira , Kappler John , Marrack Philippa 
Published: January 2023

Abstract
Besides secretion of antigen-specific antibodies, B cells may play an important role in the generation of immune responses by efficiently presenting antigen to T cells. We and others recently described a subpopulation of CD11c+ B cells (Age/autoimmune associated B cells, ABCs) which appear with age, during virus infections and at the onset of some autoimmune diseases and which participate in autoimmune responses by secreting autoantibodies. Here we assessed the ability of these cells to present antigen and activate antigen-specific T cells. We demonstrated that ABCs present antigen to T cells, in vitro and in vivo, better than follicular B cells (FO cells) do. Our data indicate that ABCs express higher levels of the chemokine receptor, CCR7, and have higher responsiveness to CCL21 and CCL19 than FO cells and are localized at T/B cell border in spleen. Using multiphoton microscopy we have shown that in vivo, CD11c+ B cells form significantly more stable interactions with T cells than Follicular B cells. Together these data identify a previously undescribed role for ABCs as potent antigen-presenting cells and suggest another potential mechanism by which these cells can influence immune responses and/or the development of autoimmunity.
Concordance of increased B1 cell subset and lupus phenotypes in mouse and human dependent on BLK expression genotypes (BA3P.114)
Authors: Wu Ying-Yu , Georg Ina , Díaz-Barreiro Alejandro , Varela Nieves , Lauwerys Bernard , Kumar Ramesh , Bagavant Harini , Castillo-Martín Mireia , Salem Fadi , Marañón-Lizana Concepción , Alarcón-Riquelme Marta 
Published: January 2023

Abstract
Polymorphisms in the BLK gene have been associated with autoimmune diseases, including systemic lupus erythematosus (SLE), with risk correlating with reduced expression of BLK. How reduced expression of BLK causes autoimmunity is unknown. Using Blk+/+, Blk+/-, and Blk-/- mice, we show that aged female Blk+/- and Blk-/- mice produced higher anti-dsDNA IgG antibodies and developed immune complex-mediated glomerulonephritis, compared to Blk+/+ mice. Starting at young age, Blk+/- and Blk-/- mice accumulated increased numbers of splenic B1a cells, which differentiated into class-switched CD138+IgG-secreting B1a cells. Increased infiltration of B1a-enriched cells into the kidneys was also observed in aged Blk+/- and Blk-/- mice. In human, we found that healthy individuals had BLK genotype-dependent levels of anti-dsDNA IgG antibodies as well as increased numbers of a B1-like cell population, CD19+CD3-CD20+CD43+CD27+, in peripheral blood. Furthermore, we describe the presence of B1-like cells in the tubulointerstitial space of human lupus kidney biopsies. Taken together, our study reveals a previously unappreciated role of reduced BLK expression on extraperitoneal accumulation of B1a cells in mice, and the presence of IgG autoantibodies and B1-like cells in human.
MH7A human synovial cell migration is dependent on the expression of B cell activating factor (BA3P.115)
Authors: Lee Jiyoung , Lee Jiyoung , Park So-Jin 
Published: January 2023

Abstract
Rheumatoid arthritis (RA), one of autoimmune diseases, is characterized by histological hallmarks such as hyperplasia of the synovial lining cells, mononuclear cell infiltration, angiogenesis, and osteoclastic bone destruction. Synoviocyte migration and invasion play a role in the chronicity and severity of RA. B cell activating factor (BAFF) which plays a role in maturation and maintenance of B cells is also associated with autoimmune RA. However, limited information is available on the effect of BAFF on synoviocyte migration. Here, we investigated whether BAFF could regulate synoviocyte migration in the presence of absence of TNF-a, the main pro-inflammatory cytokine in RA using MH7A synovial cells that is established by transfection with the SV40 T antigen (Miyazawa et al., 1998). Cell migration was assessed using Boyden chamber assay. When BAFF expression was inhibited with BAFF-siRNA, MH7A cell migration was decreased gradually in the presence of absence of TNF-. While BAFF expression was dependent on p38 kinase activation, no changes in BAFF expression was detected by the inhibition of JNK phosphorylation. In conclusion, our results suggest for the first time that migration of synovial cells in RA could be controlled by BAFF expression.
Differential responses to autoantibody deposition in CD45E613R BALB/c and C57Bl/6-129 F1 glomeruli is mediated by alterations in the autoantibody repertoire and inflammatory renal mononuclear cells (BA3P.110)
Authors: Hai Si-Han , Hermiston Michelle 
Published: January 2023

Abstract
The mechanisms regulating the transition from autoreactivity to autoimmunity are poorly understood. CD45E613R mice express a point mutation in the juxtamembrane wedge that results in dysregulated phosphatase activity. Despite similar hyperactive ITAM-mediated signaling in multiple cell lineages, the phenotypic consequences of this mutation vary with genetic context. While both BALB/c and C57Bl/6-129 F1 CD45E613R mice develop similar high titer anti-nuclear antibodies (ANA) and immune complex deposition in the glomeruli, only F1 mice develop glomerulonephritis (GN). Here, we leverage these distinct phenotypes in the context of a common point mutation to define mechanisms leading to GN. BALB/c ANA are primarily anti-dsDNA while F1 ANA are anti-dsDNA, -H3 and -Sm/RNP. Consistent with this profile, F1 B cells have increased TLR7/9 signaling. Treatment of RAW264.7 cells with aged F1 but not BALB/c CD45E613R sera induces an inflammatory phenotype. Additionally, IL-10, IL-12, MCP-1 and RANTES are elevated in aged F1 CD45E613R mice. Analysis of young, autoantibody-free mice shows increased CD11b+ CD11c+ F4/80+ renal mononuclear cells surrounding F1 but not BALB/c CD45E613R glomeruli. Upon stimulation, only F1 CD45E613R renal mononuclear cells produce TNFα and IFNγ. Together, these data support a model in which an expanded antibody repertoire combined with altered kidney immune cell subsets and altered cytokines mediate the transition from autoreactivity to autoimmunity.
Anti-insulin B cells in T-independent germinal centers (BA3P.111)
Authors: Williams Jonathan , Henry-Bonami Rachel , Hulbert Chrys , Thomas James 
Published: January 2023

Abstract
Previous studies demonstrate B cells that harbor IgM-restricted anti-insulin transgenes are anergic. However, most pathogenic insulin autoantibodies are IgG, suggesting that class switch recombination (CSR) is an important biological process in mediating loss of tolerance and autoimmunity. This breach in the B cell compartment is widely assumed to be T cell-dependent (TD). To examine peripheral tolerance for CSR-competent anti-insulin B cells, we generated C57BL/6 mice that harbor anti-insulin VDJH-125 site-directed to the native Ig H chain locus (VH125SD). Contrary to expectations, anti-insulin B cells fail to produce IgG anti-insulin Abs following TD immunization of VH125SD mice with heterologous insulin. In contrast, anti-insulin B cells produce both IgM and IgG2a anti-insulin Abs following immunization of VH125SD mice with a T cell-independent (TI) conjugate of insulin to Brucella abortus ring test antigen (BRT). Anti-insulin B cells undergo clonal expansion in vivo and emerge as IgM+ and IgM- GL7+ Fas+ germinal center (GC) B cells following immunization with insulin-BRT, but not BRT alone. These data demonstrate that combined BCR/TLR-4 stimulation reverses anergy in anti-insulin B cells and promotes antigen-specific GC responses and production of IgM and IgG insulin autoantibodies. Environmental factors that lead to infection and inflammation could play a critical yet under-appreciated role in driving loss of tolerance and autoimmune disease.
Generation of NOD mice carrying R619W mutation in PTPN22 gene by CRISPR/Cas9-mediated genome engineering (BA4P.134)
Authors: Lin Xiaotian , Pelletier Stephane , Gingras Sebastien , Marquardt Kristi , Bottini Nunzio , Green Douglas , Sherman Linda 
Published: January 2023

Abstract
An allelic variant of protein tyrosine phosphatase nonreceptor 22 (PTPN22), PTPN22R620W, is strongly associated with type 1 diabetes (T1D) and several other autoimmune diseases in human, and increases the risk of developing T1D by 2-4 fold. Non-obese diabetic (NOD) mouse is an ideal spontaneous T1D model that shares with human many genetic pathways contributing to T1D; however, it has not previously been possible to introduce targeted mutations into NOD mice due to a lack of germline competent NOD embryonic stem cells. To overcome this technological hindrance, we used CRISPR/Cas9 technology to precisely mutate PTPN22 gene in its endogenous locus in NOD zygotes, and successfully produced 2 independent lines of NOD mice expressing PTPN22R619W variant, a murine ortholog of the human autoimmune-associated allele. We have also obtained a line carrying PTPN22 null allele. We have demonstrated the mutated PTPN22 genes in all lines have successfully germline transmitted. We have found a significant increase of T1D incidence in the heterozygous NOD PTPN22R619W mice, suggesting its strong effect in heterozygosity, which is very relevant to human T1D since PTPN22R620W is most commonly carried in heterozygosity. This is the first time the autoimmune-associated PTPN22 allele has been introduced into a spontaneous model of T1D and, to the best of our knowledge, the first time CRISPR/Cas9 technology has been used to genetically modify NOD mice.
Lupus nephritis develops in a RIP3-independent manner for both males and females (BA4P.136)
Authors: Corradetti Chelsea , Jog Neelakshi , Caricchio Roberto 
Published: January 2023

Abstract
Lupus nephritis is one of the most serious manifestations of lupus and is characterized by severe inflammation and necrosis, and if untreated leads to renal failure. Although lupus occurs more often in women, both sexes develop glomerulonephritis, which in males develops earlier and is more severe than in females. Previous work by our lab found that loss of Poly [ADP-ribose] polymerase 1 (PARP-1) activity, an enzyme involved in DNA repair and necrotic cell death, resulted in milder nephritis and higher survival rates among males only. A second necrotic pathway involves Receptor-Interacting Serine-Threonine Kinase 3 (RIP3), hence in this study we aimed to investigate the impact of RIP3 on nephritis and determine if this pathway is relevant in both sexes. To this end we used three inducible murine models of lupus nephritis: nephrotoxic serum (NTS)-induced nephritis, pristane-induced lupus and chronic graft versus host (cGVH) disease. We found that absence of RIP3 imparts no positive or negative effects on the development of lupus nephritis in all three models and in both male and female mice. In addition, in vivo experiments done in absence of both RIP3 and PARP-1 did not result in any additive effects between the two pathways, conferring protection only when PARP-1 was absent. Based on these studies, we conclude that RIP3 does not appear to play a significant role in lupus nephritis, nor does it appear to interact with PARP-1 to accelerate, worsen or ameliorate the disease.
STAT4 controls GM-CSF production by both Th1 and Th17 cells during EAE (BA4P.140)
Authors: Harrington Laurie , Nozell Susan , Rajbhandari Rajani , Benveniste Etty , McWilliams Ian 
Published: January 2023

Abstract
Factors mediating Th1 and Th17 CD4 T cell driven autoimmunity remain unclear. In the mouse model of Multiple Sclerosis, experimental autoimmune encephalomyelitis (EAE), mice genetically deficient in STAT4 are resistant to disease. In contrast, deletion or inhibition of the Th1-associated cytokines IL-12 or IFNγ, which act upstream and downstream of STAT4, respectively, does not ameliorate disease. This highlights a non-canonical role for STAT4 during EAE. Recent reports have indicated that GM-CSF, a cytokine produced by both Th1 and Th17 effector CD4 T cell subsets, is critical for EAE pathogenicity. Importantly, the requirement for STAT4 in mediating GM-CSF production has not yet been investigated. Herein, we demonstrate that STAT4 controls GM-CSF production by both Th1 and Th17 CD4 T cells during EAE, and this is CD4 T cell intrinsic. Furthermore, STAT4 mediates CD4 T cell derived GM-CSF production independently of the cytokines IL-12 and IL-23. Importantly, we show that STAT4 directly interacts with the Csf2 locus in effector CD4 T cells during EAE. Overall, these studies illustrate the previously unrecognized role of STAT4 to regulate GM-CSF production by both Th1 and Th17 effector cell subsets during EAE pathogenesis.
Induction of arginase-1 by TNF-α in oligodendrocytes is controlled by SHP-1 (BA3P.119)
Authors: Minchenberg Scott , LaRocca Daria , Massa Paul 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is a debilitating neurological disease characterized by inflammatory demyelination in the central nervous system (CNS), and tumor necrosis factor (TNF-α) has been implicated in its pathogenesis. TNF-α is toxic to the myelin forming oligodendrocytes and their connected myelin sheaths, and its expression is elevated in MS lesions. However, the precise mechanism for TNF-α-mediated toxicity to oligodendrocytes remains unknown. Moreover, the role of regulatory molecules, including SHP-1, in controlling both TNF-α signaling and susceptibility to TNF-α-mediated demyelinating diseases in the CNS is unclear. Here, we show that compared to normal littermate controls, oligodendrocytes of SHP-1-deficient mice (motheaten), which are highly susceptible to inflammatory demyelination, display a significant increase in inflammatory gene expression in response to TNF-α. Unexpectedly, the most highly induced of these inflammatory genes is arginase-1, an enzyme that converts arginine to L-ornithine with major effects on allergic-type immune responses. To elucidate the mechanism underlying this induction, we are determining how TNF-α induces transcriptional activation of the arginase-1 promoter in an SHP-1-dependent manner. Further, we are investigating if abnormal expression of arginase-1 in oligodendrocytes is responsible for the oligodendrocyte dysfunction and increased demyelination observed in SHP-1-deficient mice.
Chronic liver fibrosis triggers autoantibody production by B cells in a MyD88-dependent pathway (BA4P.126)
Authors: Thapa Manoj , Tedesco Dana , Elrod Elizabeth , Han-Hwan Jin , Grakoui Arash 
Published: January 2023

Abstract
Our recent finding suggests that B cells contribute to liver fibrosis through hepatic stellate cells (HSCs) activation. While antibodies produced by the activated intrahepatic B cells contribute to autoimmunity is not known, mixed cryoglobulinemia and other B cell disorders are the leading extrahepatic manifestations of advanced liver diseases in humans. Here, our aim was to determine whether increased antibody production during liver fibrosis could potentially lead to systemic autoimmunity by production of autoantibodies and immune complexes (IC). We studied the role of B cells in autoimmunity using a carbon tetrachloride (CCl4)-induced mouse model of fibrosis and a genetically engineered multidrug resistance gene 2 knock out (Mdr2-/-) mice, which constitutively develop progressive liver fibrosis. In both fibrosis models, the frequency of B cells in the liver was elevated and B cells exhibited increased state of activation as measured by CD44 and CD86 expressions, constitutive IgG production and elevated TNF-a production. By using antinuclear antibody (ANA) staining with HEp-2 stabilized substrate and C1q standard ELISA, we detected the presence of antinuclear antibodies (ANA) and IC respectively in the serum of C57BL/6 mice treated with CCl4 for six weeks and Mdr2-/- mice at 25 week. Interestingly, the targeted deletion of MyD88 signaling in B cells during liver fibrosis resulted in reduced ANA in the serum suggesting the critical role of MyD88 in systemic autoimmunity.
Regulation of chronic lung inflammation to house dust mite allergen by Wnt antagonist (CAM1P.144)
Authors: Bothwell Alfred , Chae WookJin , Teixeira Alexandra , Chan Pamela , Hao Liming , Rothlin Carla , Krause Diane 
Published: January 2023

Abstract
Recruitment of leukocytes in response to allergen is an important step in the initiation of airway inflammation. Here we demonstrate that the Wnt antagonist Dkk-1 regulates the formation of leukocyte-platelet aggregates (LPAs) following house dust mite (HDM) allergen challenge. We show that circulating Dkk-1 is elevated after recurrent HDM allergen challenges and the source is activated platelets. The increase of P-selectin glycoprotein ligand-1 (PSGL-1) expression in leukocytes was mediated by Dkk-1 while other cell adhesion molecules such as ICAM-1 or LFA-1 were only marginally affected. The formation of LPAs was markedly reduced in Dkk-1 hypomorphic doubleridge (Dkk-1d/d) mice that had a 90% reduction of circulating Dkk-1. Recurrent allergen challenges in Dkk-1d/d mice showed significantly reduced infiltration of leukocytes including neutrophils in the lung compared to wildtype mice. Importantly, we show that type 2 cytokines (e.g. IL-4, IL-5 and IL-13) from CD4 T cells in mediastinal lymph nodes (med LNs) of Dkk-1d/d mice were substantially reduced compared to wildtype mice following allergen exposure. Consistently, we observed Gata-3 and c-Maf induction in Th2 cells did not occur in med LN CD4 T cells from Dkk-1d/d mice. Therefore, our results demonstrate that the Wnt antagonist from activated platelets regulates the early stage of leukocyte infiltration in the challenged tissue, thereby regulating inflammation and type 2 immune responses.
A novel IL233 fusion cytokine in a therapeutic approach for acute kidney injury (CAM1P.155)
Authors: Stremska Marta 
Published: January 2023

Abstract
Since we found that interleukin (IL)-2 upregulated IL-33 receptor-ST2 on CD4+ T-cells, we hypothesized that IL-2 and IL-33 may cooperate to expand endogenous Tregs. Indeed Tregs expressed ST2 and IL-33 contributed to increase Treg-expansion. We designed a fusion cytokine (IL233) containing the activities of IL-2 and IL-33 for better targeting of Tregs. We expressed the cytokines in E.coli, purified them to homogeneity and tested them in mouse model of ischemia reperfusion injury (IRI). C57BL/6 male mice were pretreated (ip) with different doses of cytokines prior to 26 min of bilateral ischemia and 24 hours of reperfusion and kidneys were characterized for function (plasma creatinine) and acute tubular necrosis. IL-2 and IL-33 in combination or as IL233 fusion cytokine, but not alone protected mice from IRI, with IL233 being more efficient than the combination. Our data suggests that the IL233 treatment first increases splenic levels of Tregs followed by their mobilization. Since both IL-2 and IL-33 also promote T-helper (Th)-2 response, an increase in IL-4 and IL-5, and a decrease in inflammatory cytokines IFN-γ and TNF-α were observed, indicating a shift towards a protective Th2 response. Adoptive transfer studies revealed higher degree of protection in mice injected with Tregs obtained from mice pretreated with IL233. Thus, IL233 fusion cytokine attenuates kidney inflammation to protect from IRI and bears strong potential to be a therapeutic agent for acute kidney injury.
Inhibitory effect of serum amyloid A on receptor activation of nuclear factor κB ligand-induced osteoclast formation (CAM1P.160)
Authors: Bae Yoe-Sik , Oh Eunseo , Lee Ha Young , Kim Hak Jung , Park Yoo Jung 
Published: January 2023

Abstract
Serum amyloid A (SAA) is a major acute-phase protein produced from the liver. Since the level of SAA rapidly increases up to approximately 1,000-fold during inflammation, it has been known as a marker of several chronic inflammatory diseases such as atherosclerosis. In this study, we find that SAA strongly inhibits osteoclast formation induced by RANKL. SAA not only markedly blocked the expression of several osteoclast-associated genes including TRAF6 and OSCAR but also strongly induced the expression of negative regulators such as MafB and IRF-8. Moreover SAA decreased c-fms expression levels on the cell surface during osteoclastogenesis via shedding of the c-fms extracellular domain. SAA also restrained fusion of osteoclast precursors by blocking intracellular ATP release. This inhibitory response of SAA is not mediated by well-known SAA receptors such as formyl peptide receptor 2, Toll-like receptor 2, or Toll-like receptor 4. These findings provide insight into a novel inhibitory role of SAA in osteoclastogenesis, and suggest that SAA is an important endogenous modulator which regulates bone homeostasis.
Anti-inflammatory effects of essential oils extracted from Chamaecyparis obtusa in inflammatory models of mice and raw 264.7 cells (CAM1P.165)
Authors: Park Yujin , Yoo Seung-Ah , Kim Wan-uk , Cho Chul-Soo , Yoon Chong-Hyeon 
Published: January 2023

Abstract
Although the biological activities of essential oil extracted from Chamaecyparis obtusa (EOCO) are not yet fully understood, antimicrobial, antifungal and anti-inflammatory effects have been reported. In this study, we investigated the anti-inflammatory effects of EOCO in two inflammatory models of mice, carrageenan-induced paw edema and thioglycollate-induced peritonitis, and lipopolysaccharide (LPS)-stimulated Raw 264.7 macrophage cells. Levels of pro-inflammatory cytokines were analyzed by ELISA, expression of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were determined by western blotting, and nitrite concentration was measured using Griess reagent. The thicknesses of mice paw edema induced by carrageenan and interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels in paw homogenates were significantly decreased in the EOCO (5 and 10 mg/kg) group as compared with control group. In mice with thioglycollate-induced peritonitis, EOCO (5 and 10 mg/kg) treatment reduced the numbers of total cells and suppressed tumor necrosis factor (TNF)-α, IL-1β, and IL-6 levels in peritoneal fluid. EOCO also decreased nitric oxide (NO), TNF-α and IL-6 production and inhibited iNOS and COX-2 expressions in LPS-stimulated Raw 264.7 cells. These data showed that EOCO has anti-inflammatory effect both in vivo and in vitro, and this effect may be related with inhibition of inflammatory mediators. In conclusion, EOCO could be an effective therapeutic agent in inflammatory diseases.