bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interferon alpha causes SLE by expanding CD4- CD8- double negative T cell (44.25)
Authors: Honda Eriko , Akiyama Chieri , Hashiramoto Akira , Shiozawa Shunichi 
Published: January 2023

Abstract
Objective: It has been hypothesized that high interferon alpha (IFNα) levels cause systemic lupus erythematosus (SLE), however, the direct proof for this is lacking. We have shown that sole increase of interferon alpha (IFNα) in doxycycline (Dox)-inducible transgenic (IFNα Tg) mice induces production of anti-dsDNA antibody and renal disease akin to human SLE. In this study, we have investigated how IFNα functions and to what extent IFNα operates in the pathogenesis of SLE. Methods: We made C57BL6/FVBN TetOp-mIFNα/EμSR-tTA transgenic female mice in which IFNα was expressed after the cessation of Dox. We transferred CD4+ (5x106), CD8+ (5x106) and CD4- CD8- double negative (DN) T (3x105) subsets derived from 30 weeks Dox-off IFNα Tg mice into naive recipient mice twice. In additional experiment, CD4+ (8x106), CD8+ (8x106) and DN T (9x105) subsets were transferred. Anti-dsDNA antibody, serum IC, proteinuria and renal lesion of recipient mice were evaluated. Results: Transfer of DN T (3x105) subset of Dox-off IFNα Tg mice induced IC-mediated mesangio-proliferative glomerulonephritis. However, there were no increases in anti-dsDNA antibody, serum IC nor proteinuria after the transfer of DN T (9x105) subset. It was, however, noted that alopecia, was developed by transfer of DN T (9x105) subset. Conclusion: DN T cell is responsible for the induction of lupus glomerulonephritis and alopecia.
DDE and diet-induced obesity alter thymocyte and splenocyte immune cell populations and renal expression of osteopontin and MCP-1 in a murine model (44.31)
Authors: Viselli Susan , McKenna Ryan , Hillhouse David 
Published: January 2023

Abstract
DDE, a metabolic breakdown product of the pesticide DDT, accumulates in adipose tissue and acts as an anti-androgen. Since androgens can protect against autoimmuity, DDE may impact autoimmmune developement. We predicted greater effects of DDE in mice with more adipose tissue. Male C57BL/6J mice were fed either a high-fat (60 kcal% fat) or control low-fat(10 kcal% fat) diet from weaning. At 8 weeks, mice were treated with DDE (200 mg/kg body wt)or vehicle by oral gavage. Mice were sacrificed at 10 weeks or 6 months. Body weights were at least 50% greater in high-fat diet mice. Total spleen and thymus cell numbers were not altered by diet or DDE. Fewer mature thymus T cells (CD3+) were found in the DDE high fat group compared with the DDE low fat group (25.22 +/- 2.76 % vs. 47.55 +/- 3.69%, p&lt;0.05) at 10 weeks. High fat diet, independent of DDE treatment resulted in more CD8+ single-positive thymocytes in high fat controls compared with low fat controls (2.04 +/- 0.9 % vs. 1.13 +/- 0.13%, p&lt;0.05) at 10 weeks and more CD4+ splenocytes in high fat compared with low fat controls (25.63 +/- 2.67% vs. 16.30 +/- 0.79%, p&lt;0.05) at 6 months. DDE caused a 10-15% increase in lymphocytic infiltrates in kidney glomeruli regardless of diet. High fat diet and DDE treatment increased renal expression of MCP-1 while DDE treatment increased OPN expression. In conclusion, diet and obesity influence some effects of DDE on the immune system. Study of additional immune parameters is warranted.
BLyS rescues RF producing B cells from activation induced death (44.34)
Authors: Oropallo Michael , Kiefer Kerstin , Marshak-Rothstein Ann , Cancro Michael 
Published: January 2023

Abstract
Rheumatoid factor (RF) autoantibodies are associated with Systemic Lupus Erythematosus and other autoimmune disorders. Previous research using AM14 mice, which have a transgene encoding a B cell antigen receptor (BCR) typical of pathogenic RF producing B cells, has demonstrated that these cells are induced to divide by chromatin immune complexes (chromatin-ICs). This work revealed a combined requisite for BCR and Toll-like receptor (TLR) signaling in the onset of RF production., We have recently found that although AM14 B cells stimulated with chromatin-ICs indeed divide, they die rapidly within 48-72 hr of the initial stimulus; by contrast, anti-IgM, CpG, or anti-IgM+CpG stimulated cells continue to proliferate within this time frame. Importantly, we have found that the cytokine BLyS (B lymphocyte stimulator) rescues survival of AM14 B cells proliferating in response to chromatin-ICs. We believe our observations may link the immune complex mediated activation of RF producing B cells with the finding that elevated BLyS levels are often seen in SLE. The ability of BLyS to rescue AM14 B cells may involve signaling through either TACI or BR3, as we have found that both of these BLyS binding receptors are upregulated after chromatin-IC stimulation. Finally, we have begun to examine the apoptotic pathway downstream of chromatin-IC stimulation, and have found that it is cell intrinsic and involves caspases 9, 3, and PARP.
Role of bone marrow-derived cells and inflammation in spontaneous cardiac dystrophic calcification (44.36)
Authors: Glass Aaron , Darwish Nedal , Taffet Steven 
Published: January 2023

Abstract
Dystrophic calcification (DC), or extra-osseous deposition of calcium phosphate, is associated with numerous cardiovascular diseases ranging from acute conditions such as myocardial infarction to chronic conditions including atherosclerosis. Development of DC in mouse models is multifactorial, with diet, age and genotype each contributing to the disease phenotype. Although DC has been characterized in a several mouse strains, the relationship between inflammation and DC has not been addressed and no autoimmune component has been identified. We identified a substrain of BALB/c mice (BALB/cByJ) that spontaneously develop epicardial DC lesions containing focal deposits of calcium and infiltrating mononuclear cells. Bone marrow chimera experiments using GFP-expressing donor bone marrow revealed that the lesions are strongly GFP positive, suggesting an inflammatory component of the disease. In particular, there was substantial GFP positive cell infiltration at the site of calcinosis. A characterization of the infiltrating cells by immunohistochemistry confirmed the presence of immune cells. Initiation of experimental autoimmune myocarditis in these animals led to the formation of a more pronounced lesion, where the calcinosis appeared to serve as a focus of the myocarditis. Further study of this model may help elucidate the dynamic interplay between inflammation and calcification in cardiovascular disease.
Antigenic targets in Aire-mediated ovarian autoimmunity (44.41)
Authors: Cheng Mickie , Fan Una , Johannes Kellsey , Anderson Mark 
Published: January 2023

Abstract
Ovarian autoimmunity is a poorly understood cause of premature ovarian failure and infertility among women. Diagnosis of autoimmune ovarian disease (AOD) is challenging due to the lack of specific and sensitive autoantibody tests, and the disease pathogenesis and antigens targeted in AOD remain largely unknown. We have established a novel mouse model of AOD using the Aire-deficient mouse. Like APS1 patients that lack AIRE, Aire-deficient mice develop disease of multiple organs, including the ovary, due to a breakdown in central tolerance. Aire-deficient female mice develop autoimmune oophoritis manifested by 100% incidence of histologic disease and frequent oligoclonal autoantibodies to ovarian tissue, presenting a valuable spontaneous disease model in which to identify ovarian antigens. Initial investigations have identified putative antigenic targets, including a 60 kd ovarian antigen that is targeted early in AOD. Analysis of ovarian infiltrates in Aire-deficient females indicates a central role for Th1 CD4+ T cells in mediating disease. Similar autoantibody profiles and immune infiltrates have also been reported in women with premature ovarian failure, suggesting that common ovarian antigens may be shared. Understanding of antigens and disease pathogenesis in Aire-mediated AOD will define the mechanism by which tolerance is broken in the ovary and will have potential for translation to patients in a significant cause of human ovarian failure and infertility.
Autoreactive Tbet-positive CD4 T cells develop independent of classic Th1 cytokine signaling during EAE (44.45)
Authors: Yeh Wen-I , Harrington Laurie 
Published: January 2023

Abstract
Many autoimmune diseases are associated with the presence of Th1 and Th17 effector CD4 T cells. Paradoxically, the principal Th1 cytokine IFNγ does not appear necessary for disease, but the key Th1-associated transcription factor Tbet is required. This conundrum propelled us to investigate the regulation of this transcription factor during autoimmunity. Following the onset of experimental autoimmune encephalomyelitis (EAE), we observed a preferential upregulation of Tbet by CD4 T cells within the CNS, but not the secondary lymphoid organs. These Tbet-positive CD4 T cells were capable of producing the cytokine IFNγ, and a proportion of these cells produced both IFNγ and IL-17A. Interestingly, these Tbet-positive cells were present in high frequencies during disease in IFNγ-deficient mice. Moreover, we found that CD4 T cells from IFNγ-deficient/IFNγ reporter mice upregulated the Thy1.1 reporter, indicating the presence of Th1 or “Th1-like”, Tbet-positive CD4 T cells even in the absence of the cardinal Th1 cytokine, IFNγ. These data support the concept that Th1 cells contribute to EAE in a Tbet-dependent, IFNγ-independent manner. We further examined the requirement of other Th1-associated molecules in controlling Tbet expression during EAE, and noted that STAT1, IL-12, and IFNγ were dispensable for the induction of Tbet in vivo. Hence, this study highlights the complex regulation of Tbet and the potential unrecognized role for Th1 cells during autoimmunity.
Activating transcription factor 3 promotes sublytic C5b-9-induced glomerular mesangial cells apoptosis through up-regulation of Gadd45α and KLF6 (44.5)
Authors: Wang Yingwei , Xu Kuanfeng , Zhou Ying , Liu Xin , Qiu Wen 
Published: January 2023

Abstract
The sublytic C5b-9 complexes can result in glomerular mesangial cells (GMCs) apoptosis, which involved in the initiation and development of rat Thy-1 nephritis, but the mechanism of sublytic C5b-9-mediated GMCs apoptosis remains unclear. Activating transcription factor 3 (ATF3) is an immediate early gene for cells to cope with a variety of stress signals, and our previous study revealed that ATF3 could promote GMCs apoptosis attacked by sublytic C5b-9. However, target genes of ATF3 in GMCs apoptosis triggered by sublytic C5b-9 stimulation have not been identified. In this study, the data showed that the expression of ATF3, growth arrest and DNA damage-45 alpha (Gadd45α), Krüppel-like factor 6 (KLF6) and proliferating cell nuclear antigen (PCNA) in the GMCs induced by sublytic C5b-9 for the indicated time was significantly increased, and ATF3 expression could lead to GMCs apoptosis through up-regulation of Gadd45α and KLF6, but not up-regulation of PCNA. Furthermore, Gadd45α was identified as a downstream target gene regulated by ATF3 directly, and KLF6 might be regulated by ATF3 in an indirect manner.
A critical role of B cells in biliary disease and sialadenitis in the NOD.c3c4 model of autoimmune cholangitis. (44.8)
Authors: Moritoki Yuki , Tsuda Masanobu , Tsuneyama Koichi , Zhang Weici , Yoshida Katsunori , Lian Zhe-Xiong , Yang Guo-Xiang , Ridgway William M. , Wicker Linda S. , Ansari Aftab A. , Gershwin M. Eric 
Published: January 2023

Abstract
The role of B cells in the pathogenesis of PBC in humans has been a controversial problem. On the one hand, the serologic hallmark of PBC are the presence of antimitochondrial antibodies, found in &gt;90% of patients. However, the antibodies do not correlate with disease severity and the titers do not change following therapeutic orthotopic liver transplantation. In addition, related work on a murine model of PBC, the dnTGFβRII mice, suggests that regulatory B cells may be an important contributor to disease pathogenesis. Indeed, the absence of B cells in this model exacerbates biliary pathology. To address this problem, we have taken advantage of the availability of both dnTGFβRII mice and NOD.c3c4 mice, to study the role of B cells in the natural history of disease. In particular, we generated genetically B cell deficient (Igμ-/-) NOD.c3c4 mice and compared the immunopathology of these mice to control B cell sufficient (Igμ+/+) NOD.c3c4 mice. Igμ-/- NOD.c3c4 mice not only had an amelioration of salivary gland inflammation, but also reduced numbers of inflammatory liver infiltrates, ameliorated liver inflammation, and a significantly lower prevalence of biliary cyst formation. B cell deficient mice demonstrated decreased number of non-B cells in the liver accompanied by reduced numbers of activated natural killer cells. In conclusion, B cells play a critical role in promoting liver inflammation and cyst formation as well as salivary gland pathology in autoimmune NOD.c3c4 mice.
FCRLA is an intracellular FcR-related protein that binds IgM, IgG and IgA isotypes in B cells (45.11)
Authors: Burrows Peter , Santiago Teresa , Kulemzin Sergei , Reshetnikova Evdokia , Chikaev Nikolai , Volkova Olga , Mechetina Ludmila , Zhao Meina , Davis Randall , Taranin Alexander , Najakshin Alexander , Hendershot Linda 
Published: January 2023

Abstract
Fc-receptor-like A (FCRLA) is a novel member of the extended Fc receptor (FcR) family. Unlike the classical FcRs and Fc receptor-like (FCRL) molecules, which are typically transmembrane receptors with extracellular ligand binding domains, FCRLA has no predicted transmembrane domain or N-linked glycosylation sites and is positioned intracellularly. We show by confocal microscopy and biochemical assays that FCRLA is a soluble, resident ER protein; however, it lacks a characteristic KDEL retention sequence at its C-terminus. Using a series of deletion mutants, we found that its ER retention is most likely mediated by the amino terminal domain. We have also recently identified ER localized immunoglobulin as an FCRLA ligand. These results indicate that FCRLA is unique among the large FcR family in that can associate with multiple immunoglobulin isotypes, including IgM, IgG, and IgA. To better understand the role of this protein in human disease, B cell chronic lymphocytic leukemia (CLL) samples were assayed for FCRLA expression by flow cytometry. Interestingly, the receptor was more highly expressed by the indolent form of CLL, which is distinguished by somatically hypermutated heavy chain variable region genes, than by the aggressive high-risk form, which features more germline B cell receptor characteristics. These results indicate that FCRLA may thus be a useful diagnostic marker for CLL.
Fast, easy and column-free procedure for the isolation of human memory B Cells (45.14)
Authors: Fairhurst Maureen , Yu Jessie , Thomas Terry 
Published: January 2023

Abstract
Human memory B cells have the unique ability to rapidly proliferate and differentiate when re-exposed to the same antigen. They can be distinguished from naïve B cells by the presence of somatic hypermutations in their Ig-V region sequences. CD27 is widely used as a marker of memory B cells because its expression correlates with the presence of such mutations. Several CD27+ memory subsets collectively make up 20-60% of peripheral B cells. Using a two-step EasySep™ method, memory CD27+ B cells are isolated from fresh or previously frozen peripheral blood nucleated cells. First, non-B cells are targeted for depletion with dextran-coated magnetic particles using a cocktail of tetrameric antibody complexes (TAC). Labeled cells are separated in an EasySep™ magnet without the use of columns and pre-enriched, unlabeled B cells are collected. Next, CD27+ B cells are selected from the pre-enriched fraction using TAC recognizing CD27 and dextran-coated magnetic particles. Labeled cells are separated and remain in the tube in the magnet while unwanted cells are poured off. The unwanted cell fraction may be used to obtain naïve B cells. The selection steps can be fully automated using RoboSep™. Starting with a frequency of 4.1 ± 1.2% CD19+CD27+ B cells, purities of 93.0 ± 3.4% (n=14) can be obtained. Isolation of memory B cells from human samples is increasingly important for the investigation of B cell signaling pathways and regulation mechanisms central to a robust immune response.
Sustained antibody responses are dependent on intrinsic CD28 function in plasma cells (45.15)
Authors: Rozanski Cheryl , Arens Ramon , Carlson Louise , Nair Jayakumar , Boise Lawrence , Chanan-Khan Asher , Schoenberger Stephen , Lee Kelvin 
Published: January 2023

Abstract
Sustained long-term antibody (Ab) levels are the cornerstone of protective immunity, yet it remains unclear how they are maintained. A predominant theory is this occurs via antigen-independent Ab production by a subset of long-lived plasma cells (LLPC) that survive within bone marrow (BM). However, the model's central tenets that BM LLPC are a separate subset defined by intrinsically distinct biology and contribute to long-term Ab titers, have not been demonstrated. We now report that long-term humoral immunity is dependent on plasma cell-intrinsic function of CD28, which selectively supports the survival of BM LLPC but not splenic short-lived PC (SLPC). LLPC and SLPC both express CD28, but CD28 signaling with enhanced survival occurred only in the LLPC. Addition of BMDC improved the survival of WT LLPC 2-fold over that seen with media alone and resulted in a significant increase in IgG production; in contrast to CD28-/- PC when cocultured with BMDC. This suggests a direct role for CD28 in PC-DC interaction. In vivo, even with sufficient T cell help the loss of CD28 or its ligands CD80/CD86 caused significant loss of the LLPC population, reduction of LLPC survival t1/2 from 426 to 63 days, and inability to maintain Ab titers long-term - with no effect on SLPC populations. These findings establish the existence of the distinct BM LLPC subset that is necessary to sustain Ab titers, and uncover a central role for CD28 function in the longevity of PC and humoral immunity.
Host mediated modulation of autoreactive B cell differentiation (45.5)
Authors: Kiefer Kerstin , Marshak-Rothstein Ann 
Published: January 2023

Abstract
Selection of B cells into the mature B cell compartments is based on the affinity of the B cell receptor (BCR) for self-ligands. Strongly self-reactive B cells undergo receptor editing or deletion during this process to avoid autoimmunity. Current dogma suggests that B cells expressing BCRs with low affinity to self become follicular (FO) B cells whereas B cells with intermediate affinity to self develop into marginal zone (MZ) B cells. We have examined this selection process using AM14 BCR gene-targeted mice on either a BALB/c or BALB/c x C57BL/6 F1 background. In BALB/c mice, all mature AM14 B cells move into the FO compartment, are fully functional and respond normally to BCR crosslinking. In contrast, on a BALB/c x C57BL/6 F1 genetic background, AM14 B cells move into the MZ as well as the FO compartment. Importantly, these AM14 B cells appear anergic and do not respond well to BCR crosslinking. In vitro, we found that BALB/c AM14 B cells proliferate in response to RAG-deficient C57BL/6 spleen cells, but not to C57BL/6 cells from the bone marrow or peritoneal cavity. These findings suggest that a ligand expressed by splenic non-T/non-B C57BL/6 cells can modulate the differentiation pathway of AM14 B cells. To further investigate which cell type expresses this self-ligand, we are comparing AM14 B cell engraftment of BALB/c and C57BL/6 F1 mice using mixed bone marrow chimeras. The results of these studies and the implications for MZ B cell selection will be discussed.
Activation induced cytidine deaminase is highly expressed on CD99+Faslow Ramos cells; implication in the early centroblast stage (45.8)
Authors: Lee Hyun Gyu , Park Eunsun , Lee Jae Myun , Park Jeon Han 
Published: January 2023

Abstract
Ramos cell line, derived from Burkitt’s lymphoma with centroblastic phenotype, has been chosen as a good model for studying somatic hypermutation. To delineate maturation stages of B cells in germinal center (GC) with an expression pattern of CD99 that has been reported to be down-regulated in GC, we sorted and subcloned the Ramos cells according to the expression patterns of CD99. CD99+ clones lost CD99 expression to become CD99- cells spontaneously. However, the reverse event has not happened in any cases. We observed that CD99+ clones have earlier centroblastic phenotypes than CD99- ones through experiments such as IgM-induced apoptosis and Fas expression. When the CD99+ clones were subcloned based on Fas expression levels, CD99+Faslow clones generated IgM- population in higher frequency than CD99+Fashi and CD99-Fashi did, then we evaluated the expression level of AID and found that CD99+Faslow clones are the cells at the stage where AID level is the highest. This high expression level of AID in the CD99+Faslow clones is pertinent to the Ramos cells sorted directly and also to different Burkitt’s lymphoma cell lines: Daudi, BJAB, and Raji. These results suggest that AID is differentially expressed between CD99+ and CD99- cells, implying CD99 might be a marker to delineate the maturation stages of B cells in GC. Based on this, we hypothesize that CD99+Faslow cells are immature cells and differentiate into CD99-Fashi with an intermediate step which has CD99+Fashi in GC.
CD99+Faslow Ramos cells, representative of early centroblasts, are sensitive to IgM induced apoptosis (45.9)
Authors: Park Kwang Hwan , Lee Hyun Gyu , Park Eunsun , Lee Jae Myun 
Published: January 2023

Abstract
Ramos cell line, derived from human Burkitt’s lymphoma, expresses a centroblastic phenotype in the early stage of B cell development in germinal center. In previous study, CD99 positive cells were scattered throughout the lymph nodes, but most cells in germinal center had low expression of CD99 in immunohistochemical staining. Here, we hypothesize that CD99+Faslow Ramos cells may be representative of early centroblasts and have sensitivity to IgM induced apoptosis. Using B cells from human tonsils, we have evaluated various markers of centroblastic stage. We found that the expression of Fas was low in the early centroblastic stage and gradually increased during differentiation. Ramos cells that have heterogenous expressions of CD99 were subcloned into CD99+ and CD99-. IgM induced apoptosis in CD99+ clones was higher than that of CD99- clones. CD99+ clones showed heterogeneous patterns of Fas expression from low to intermediate levels, while CD99- clones were homogenous in terms of intermediate expression of Fas. CD99+Faslow and CD99-Fashigh clones were subcloned, and the expression of Fas was gradually increased during the long term culture of CD99+Faslow clones. CD99+Faslow clones were the most sensitive cells for IgM induced apoptosis. All of these results indicate that CD99+Faslow cells may be representative of early centroblasts and sensitive to IgM induced apoptosis. we will investigate the molecular mechanism of this apoptotic signaling pathway in the early centroblast.