bluuebunny/crossref_metadata_2025_split huggingface dataset:
Contribution of VH Replacement Products to the Generation of Autoreactive Antibodies in Systemic Lupus Erythematosus (129.4)
Authors: Fan Run , Su Kaihong , Majors Ashley M , Yu Zhihong , Zhao Evan , Kimberly Robert P , Zhang Zhixin 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is characterized by the production of high affinity autoantibodies against various nuclear antigens. To gain insight into the mechanisms of autoantibody generation during the course of SLE, we performed single cell PCR analysis and obtained about 200 IgH genes with paired Igκ or Igλ genes from the plasma cells (CD19lowCD27high) and naïve B cells (CD19+CD27−) of three SLE patients. The SLE derived IgH genes have longer CDR3 regions with enriched Arg residues, which are the common features of autoantibodies. SLE plasma cells derived IgH and IgL genes are highly mutated, indicating their positive selection and active involvement in the abnormal immune responses. Interestingly, analyses of the IgL and IgH genes derived from SLE patients showed signs of excessive secondary recombination when compared with those from healthy donors, such as elevated usages of downstream Jκ4, Jκ5, and Jλ3 genes in the IgL genes and concurrently, significantly elevated frequencies of VH replacement products in the IgH genes derived from SLE patients. All the VH replacement “footprints” preferentially contribute charged amino acids into the IgH CDR3 regions. Initial testing of 12 recombinant antibodies derived from SLE plasma cells showed seven of them recognize peri-nuclear antigens. Importantly, the identified VH replacement products directly encode autoantibodies. These findings revealed for the first time that excessive secondary recombination occurred on both IgL and IgH genes in SLE patients and the enriched VH replacement products in SLE plasma cells directly encode autoantibodies.
IGRP-reactive T cells can be detected directly ex vivo in the blood of type 1 diabetes patients (129.45)
Authors: Jarchum Irene , Nichol Lynn , Trucco Massimo , DiLorenzo Teresa P. 
Published: January 2023

Abstract
Both CD4 and CD8 T cells are required for the development of type 1 diabetes (T1D) in NOD mice and several antigens are known to be involved in the development of the disease. Among them is IGRP (islet-specific glucose 6 phosphatase catalytic subunit related protein), which has emerged in recent years as an important antigen in NOD mice. IGRP-reactive CD8 T cells constitute up to 30–40% of islet infiltrating T cells and these cells can be detected directly ex vivo in islet infiltrates and peripheral blood. To determine whether IGRP is a CD8 T cell antigen in humans, we created a panel of 4 HLA-A2 restricted IGRP epitopes (IGRP211-219, IGRP215-223, IGRP222-230, and IGRP265-273) to be screened directly ex vivo by IFN-γ ELISPOT. We performed the study with unfractionated PBMC from 21 recent-onset T1D patients (mean age 9.2 ± 4.3 years old) and 12 healthy controls (29.7 ± 5.7 years old) with the candidate peptides. Our results show that 66% of patients and 8% of healthy controls are positive for at least 1 IGRP peptide and 57% of patients versus 0% of controls are positive for 2 IGRP peptides. These data provide evidence that IGRP is a CD8 T cell antigen in humans, and that IGRP-reactive T cells can be detected ex vivo in the blood of T1D patients. This contributes to the understanding of the underlying disease process in humans as well as to future directions for diagnosis and intervention in T1D patients.
The role of Lipocalin 2 in the pathogenesis of Lupus Nephritis (LN) (129.7)
Authors: Pitashny Milena , Qing Xiaoping , Zhao Zeguo , Putterman Chaim 
Published: January 2023

Abstract
There is strong experimental evidence that anti DNA antibodies are instrumental in LN pathogenesis. We found that exposure of murine mesangial cells to pathogenic anti DNA mAbs induces overexpression of Lipocalin 2, a protein known to be upregulated following kidney insult. In this study we explored the upregulation of Lipocalin 2 in the context of murine lupus nephritis.
Kidney mesangial and tubular cells from MRL/lpr lupus prone mice were treated with the pathogenic anti DNA mAb 1A3F; consistent upregulation of Lipocalin 2 was detected. Similarly, IL1β, an inflammatory cytokine secreted by epithelial cells and macrophages, and agonists of TLR2 and TLR4 significantly stimulated the expression of Lipocalin 2. Interestingly, we found that 1A3F binds to HMGB1, an endogenous ligand for TLR 2/4.
Old MRL/lpr mice displayed higher kidney RNA expression of Lipocalin 2 than BALB/c mice (p&lt;0.01). In addition, serum Lipocalin 2 was elevated in old as compared to young MRL/lpr mice (p&lt;0.01), and compared to old MRL/+ and BALB/c mice (p&lt;0.05). Importantly, kidney and serum Lipocalin 2 significantly correlated with anti DNA titers.
We conclude that Lipocalin 2 is upregulated in lupus kidneys, although whether this is only due to injury or plays a role in pathogenicity is yet unknown. Nevertheless, antibody deposition mediates the upregulation of Lipocalin 2 in intrinsic kidney cells with possible involvement of TLR 2/4 signaling.
Heritability plots of levels of autoantibodies in thyroiditis and type 1 diabetes using ROMP (129.8)
Authors: Outschoorn Ingrid Maive , Burek C. Lynne , Talor Monica V. , Hoffman William H , Rowley Merrill J , Mackay Ian R , Jones Timothy W , Rose Noel R 
Published: January 2023

Abstract
Few simple methods exist to study the genetic control of autoimmunity. We plotted ROMP (Regression of Offspring on Midparent) for hundreds of trios and compared affected and unaffected siblings. Heritability of autoantibodies (AAbs) to thyroglobulin and thyroperoxidase in chronic lymphocytic thyroiditis (CLT) and thyrotoxicosis (TT), and to GAD65 in type 1 diabetes (T1D) were assayed. AAbs were significantly correlated in TT (R 2 = 0.569, p = 0.001), but not in CLT or T1D. Positive results in TT were found in patients of African American origin, suggesting which quantifiable parameters may determine the sequential appearance of more than one type of antibody, as in T1D, in predicting disease onset in other family members. Parental effects can be determined separately, and data extended to epitope spreading and disease progression. Such a screening method requiring small patient numbers enable extensive studies to be rapidly carried out simultaneously and retrospectively on various AAbs.
PKCδ nitrosylation impairs ERK pathway signaling in lupus T cells (129.9)
Authors: Gorelik Gabriela , Wu Ailing , Richardson Bruce 
Published: January 2023

Abstract
T cells from lupus patients or treated with hydralazine have defective PKC-ERK pathway signaling, causing decreased DNA methyltransferase levels, DNA hypomethylation and overexpression of methylation sensitive genes contributing to autoimmunity. We asked what causes this defect. Immunoblotting mapped the defect to impaired PMA stimulated PKCδ phorphorylation in lupus and hydralazine-treated T cells (49.6 ± 4.3% decrease, p≤0.001 and 28.3± 3.2% decrease, p≤0.05 respectively). Total PKCδ levels were not affected. Rottlerin, a selective PKCδ inhibitor, decreased PMA-induced ERK pathway signaling and increased CD70 expression by demethylating its promoter similar to lupus and hydralazine treated T cells. PDK-1, which activates PKCδ, was not modified in lupus or hydralazine treated cells, and lupus serum did not inhibit PKCδ phosphorylation. This suggested an intrinsic PKCδ defect. Tyrosines in lupus proteins are aberrantly nitrosylated, so PKCδ phosphorylation was studied in peroxynitrite treated cells. Peroxynitrite caused a dose-dependent decrease in PKCδ phosphorylation (p&lt;0.05). Similarly, phospho-PKCδ was barely detectable in anti-nitrotyrosine immunoprecipitates of PMA stimulated lupus T cells, suggesting prior nitrosylation. These results suggest that defective PKCδ activation due to oxidative damage contributes to the development of lupus.
This work was supported by grants from NIH and Veterans Administration
Adoptive transfer of regulatory T cells can delay disease progression in a congenic murine model of SLE (130.12)
Authors: Cuda Carla Marie , Morel Laurence 
Published: January 2023

Abstract
Previous studies have shown that adoptive transfer of regulatory T cells (Tregs) can prevent disease onset in murine organ-specific autoimmune disease models. Our lab studies a congenic murine model of systemic lupus erythematosus containing three susceptibility loci from the NZM2410 spontaneous model bred onto a normal B6 background, called the B6. Sle1Sle2Sle3 triple congenic strain. This strain presents with a decreased number of Tregs, yet on a per-cell basis, these Tregs are equally as capable of suppressing effector T cell (Teff) proliferation as those isolated from normal controls. To determine whether the deficiency in Treg number contributed to disease, we adoptively transferred six million isolated CD4+CD25+ Tregs or CD4+CD25− Teffs from triple congenic donors into young unmanipulated triple congenic recipients. A delay in the production of anti-nuclear antibodies as well as a decrease in spleen weight, proteinuria level, and activation of effector populations were observed four months post-transfer in those mice which received Tregs. Here we show that a single injection of unmanipulated Tregs is sufficient to ameliorate observed disease phenotypes in a systemic autoimmune disease model.
This research is supported by NIH R01 AI1045050, T32 AR007603
Genetic dissection of Sle1c (130.16)
Authors: Perry Daniel James , Morel Laurence 
Published: January 2023

Abstract
The Sle1c locus, derived from the NZW mouse strain, is a ~6.5Mb interval located at the telomeric end of chromosome 1. Previous work has shown that when Sle1c is congenically bred onto a B6 background to yield B6. Sle1cw, a mild lupus-like disease occurs in aged mice (&gt;8mo.), indicated by low to moderate levels of anti-nuclear antibodies (ANA) and low incidence of glomerulonephritis (GN). Further phenotypic analyses of this strain reveal decreased T-dependant (TD) immune responses, abnormal germinal centers (GC), increased T cell activation and proliferation, amplification of chronic graft-versus-host disease (cGVHD), and decreased number of regulatory T cells (Tregs).
Cr2 is located on the telomeric end of Sle1c. It encodes Complement receptor 1 (CR1) and 2 (CR2) thorough alternative splicing has previously been described as a candidate gene for Sle1c. It was shown with subcongenic strains of Sle1c that cr2w results in the abnormal GC and decreased TD immune response phenotypes. However, cr2w does not account for the other phenotypes. We have created a series of subcongenic strains on the centromeric end of Sle1c that map the T cell activation phenotype to &lt;1Mb. Screening is ongoing for the enhanced cGVHD. This should lead to the identification of the corresponding gene(s).
This research is supported by NIH R01 AI1045050
CD8+ T Cell- mediated Suppression of Autoimmunity in a Murine Lupus Model of Peptide-Induced Immune Tolerance Depends on Foxp3 Expression (130.19)
Authors: Singh Ram P , La Cava Antonio , Wong Maida , Tamayo Alejandro M , Ebling Fanny M , Hahn Bevra H 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease caused by autoantibodies including IgG anti-DNA. NZB/NZW Fl female (BWF1) mice, a model of spontaneous polygenic SLE, tolerized with an artificial peptide (pCONSENSUS, pCons) based on anti-DNA IgG sequences containing MHC Class I and Class II T cell determinants, develop regulatory CD4+CD25+T cells and inhibitory CD8+T cells (CD8+ Ti), both of which suppress autoAb production. In the present study, using various cellular and molecular approaches, we show that inhibitory function of CD8+ T cells from tolerized mice is sustained for up to 8 weeks and at all times depends on expression of Foxp3. Both CD28− positive and –negative CD8+ T cells contain inhibitory cells, but the expression of Foxp3 and of mRNA for TGFb is higher and lasts longer in the CD28− subset. In vitro addition of TGFb (in the presence of IL-2) induces Foxp3 expression in a dose response manner. Gene inhibition or blockade with siRNA of Foxp3 abrogates the ability of the CD8+ Ti to inhibit anti-DNA production and the proliferation of CD4+ helper T cells. Moreover, we found a significant correlation between expression of Foxp3 and ability of CD8+ Ti cells to secrete TGFb. Therefore, CD8+ Ti in this system of tolerance is depend on expression of Foxp3, and there may be a bi-directional Foxp3/TGFb autocrine loop that determines the ability of the CD8+ T cells to control autoimmunity.
Supported by NIH grants AI63515,AI 63515, AR53239 and Tina C Foundation.
Activation of NKT cells exacerbates environmentally induced autoimmunity (130.23)
Authors: Vas Jaya , Mattner Jochen , Bendelac Albert , Howell Amy , Richardson Stewart , Ndonye Rachel , Monestier Marc 
Published: January 2023

Abstract
Administration of subtoxic doses of mercuric chloride to genetically susceptible mice induces an autoimmune syndrome with anti-nucleolar antibody production, polyclonal B cell activation and nephritis. We investigated the role of natural killer T (NKT) cells in the modulation of this autoimmune model. Activation of NKT cells by synthetic variants of α-galactosyl ceramide worsened manifestations of mercury-induced disease. Exposure to heat killed Sphingomonas capsulata which contains NKT cell ligands in its cell wall also worsened disease manifestations. We used this model to further elucidate the interplay between NKT cells and other immune cell types. Mice can be tolerized to mercuric chloride by administration of a very low dose of the chemical. Tolerance establishment requires antigen presentation in the steady state by dendritic cells to elicit CD4+ CD25+ regulatory T cells. Activation of NKT cells by a synthetic ligand was able to completely prevent induction, but was by itself unable to break already established tolerance. Dual stimuli (NKT cell activation and TLR 9 ligation) were required to break established tolerance. This study demonstrates that NKT cell activation worsens chemically- induced autoimmunity and also shows a synergy between NKT and TLR ligands in potentiating disease manifestations.
Supported by a NIH grant.
Invariant NKT cells exacerbate liver injury in a TGF-β receptor II dominant-negative mouse model of primary biliary cirrhosis (130.24)
Authors: Chuang Ya-Hui , Lian Zhe-Xiong , Yang Guo-Xiang , Shu Shang-An , Moritoki Yuki , Gershwin M.Eric 
Published: January 2023

Abstract
Primary biliary cirrhosis (PBC) is an organ-specific autoimmune liver disease characterized by the presence of anti-mitochondrial antibodies (AMA) and the destruction of small intrahepatic bile ducts with portal inflammation. In previous studies, we reported that both CD1d expression and the frequency of invariant natural killer T (iNKT) cells were increased in the liver of patients with PBC. To define a specific role of iNKT cells in the pathogenesis of PBC, particularly early events, we investigated the function of hepatic iNKT cells in our TGF-β receptor II dominant-negative (dnTGFβRII) mouse model of PBC. We report here that these dnTGFβRII mice demonstrate a massive increase of hyperactive iNKT cells within the hepatic tissues. We generated CD1d−/−and CD1d+/− dnTGFβRII mice and performed a comparative study of liver immunopathology, demonstrating that the CD1d−/−dnTGFβRII mice, lacking iNKT cells, exhibit significantly decreased hepatic lymphoid cell infiltrates and milder cholangitis compared to CD1d+/−dnTGFβRII mice. Interestingly, there was a significant increase in the production of IFN-γ in hepatic iNKT cells activated by α-galactosylceramide (α-GalCer) in young but not older dnTGFβRII mice, suggested an age dependent role of iNKT cells. These data demonstrate that iNKT cells in dnTGFβRII mice are a critical factor in liver injury.
INCREASED EXPRESSION OF IFN-ALPHA AND IL-6 BY HYPERACTIVE PLASMACYTOID DENDRITIC CELLS IN BXD2 AUTOIMMUNE MICE (130.25)
Authors: Wang John Hsien , Hsu Hui-Chen , Yang PingAr , Wu Qi , Job Godwin , Chen Jian , Timares Laura , Mountz John D. 
Published: January 2023

Abstract
BXD2 is a mouse model which exhibits several features of human SLE and arthritis. IFN-α is a known cytokine which promotes B-cell maturation and antibody CSR and SHM. ELISA results of in vitro culture of spleen cells demonstrates that IFN-α expression reaches a peak at 2 months of age (4-fold increase compared to nonautoimmune mice; p&lt;0.005), suggesting that IFN-α can serve as an early trigger for B-cell autoimmunity. Immunofluorescent staining for CD11c+ antigen-presenting cells identifies an increased population (2–4 fold; p&lt;0.005) of plasmacytoid dendritic cells (pDCs). Flow cytometry results demonstrate splenic pDCs are the primary cells which express IFN-α (&gt;80%) and IL-6 (&gt;96%) in BXD2 mice. In vivo treatment of 2 m-old BXD2 mice with DOTAP/CpG resulted in a 200-fold increase in IFN-α compared to a 4-fold increase in B6 mice. This is associated with high levels of autoantibody titers and CSR. In conclusion, BXD2 mice exhibit a significantly increased population of pDCs that express IFN-α at a young age which is associated with increased humoral response.
CD8+ T cells are critical for collagen induced arthritis disease progression (130.46)
Authors: Bucks Christine M , Katsikis Peter D 
Published: January 2023

Abstract
Despite extensive study of T cells in Rheumatoid Arthritis (RA), relatively little is known regarding the role of CD8+ T cells in RA progression. Here, we use the collagen induced arthritis (CIA) model, and deplete, prior to disease onset, CD8α+ T and dendritic cells (DCs) or CD8β+ T cells in collagen immunized DBA mice. Beginning 23 days post immunization, mice received repeated injections of 100μg anti-CD8α(53-6.7), or anti-CD8β(H35-17.2) or IgG2a isotype. Depletion of CD8αor CD8β cells did not affect the prevalence of disease; suggesting that CD8+ cells are not necessary for disease initiation (isotype=87%, anti-CD8α=78%,and anti-CD8β=78%, n=9/group). However, there was a significant reduction in arthritis duration with CD8α depletion (anti-CD8α= 4.9±2.0 vs. control= 8.6±1.1 arthritic days, p&lt;0.01) and with CD8β depletion (anti-CD8β= 8.1±1.7 vs. control=13.5±1.9 arthritic days). Disease severity was also reduced in CD8α and CD8β depleted mice. Depleted mice had maximum mean paw scores of 2.4±1.1 (CD8α depleted) and 2.5±1.0 (CD8β depleted) compared to 4.5±1.8 in controls, p&lt; 0.03. This was accompanied by reduced mononuclear cell infiltrate, pannus formation and joint destruction, as observed by H&E staining. We have shown that CD8+ T cells are necessary for CIA progression and are critical contributors to disease severity.
HLA-DQ6 (DQB1*0601) molecule modulate PLPp91-110 induced EAE in DR3 (DRB1*0301) mice through apoptosis. (130.6)
Authors: Mangalam Ashutosh K. , Rodriguez Moses , David Chella S 
Published: January 2023

Abstract
The HLA class II region on chromosome 6p21 accounts for majority of familial clustering in MS and is by far the major susceptibility locus. However due to multiple class II genes, high polymorphism and linkage disequilibrium, it has been difficult to understand the role of individual class II genes and interaction between class II genes in the disease process. Previously, we have shown that PLP91-110 induced classical EAE in DR3.Abo but not in DQ6.Abo transgenic (tg) mice, suggesting that DR3 is a disease susceptible gene in the context of PLP. In this study, we have generated double (tg) mice expressing HLA-DR3/DQ6 to simulate heterozygous haplotype of MS patients. Introduction of DQ6 onto DR3 tg mice led to decrease in disease incidence on immunization with PLP91-110 suggesting a protective role of DQ6. We further characterized that this protective effect was due to expression of high level of IFNγ by DQ6 restricted T cells, which suppressed proliferation of encephalitogenic DR3-restricted T cells by inducing apoptosis in antigen specific T cells. Our IFNγ blocking experiments confirmed an anti-inflammatory role of IFNγ. Our study suggests that DQ6 modifies the PLP91-110 specific T cell response in DR3 through IFNγ mediated activation induced cell death. Thus our double tg mouse model provides a novel tool to study linkage disequilibrium and explain how HLA-DQ molecule might modulate disease in MS-susceptible individual.
The role of T cell memory and apoptosis in the resistance of Drak2-deficient mice to autoimmunity (130.8)
Authors: Hernandez Jeniffer B. , Ramos Stephanie J. , Walsh Craig M. 
Published: January 2023

Abstract
Autoimmune disease often occurs via the escape of autoreactive T cells from apoptosis, implying a breakdown of central or peripheral tolerance mechanisms. We have recently demonstrated that thymocytes and T cells lacking the serine/threonine kinase DRAK2 have a diminished activation threshold, suggesting that this kinase serves as a quiescence factor to maintain tolerance to weak self-antigens. Drak2−/− T cells exhibit enhanced sensitivity to TCR mediated stimulation and produce higher levels of the cytokines IL-2, interferon gamma and IL-4. Surprisingly, Drak2−/− mice are resistant to experimental autoimmune encephalomyelitis (EAE). EAE is a disease induced in mice by immunization with MOG35-55 peptide, which elicits multiple sclerosis-like symptoms due to T cell activation and recruitment into the CNS. We have observed enhanced apoptosis in T cells derived from Drak2−/− mice, suggesting that DRAK2 serves as both a quiescence factor in naïve T cells and a survival factor in effector T cells. In order to understand the mechanism of resistance of Drak2−/− mice to EAE, we have employed adoptive transfer strategies to evaluate the T cell intrinsic requirements for DRAK2 in maintaining the survival of MOG-specific effector and memory populations during the course of EAE. These results demonstrate that DRAK2 is a key factor that regulates both central and peripheral tolerance.
Suppression of autoimmune diabetes by peptide-MHC class II dimers (131.13)
Authors: Li Li , Goudy Kevin , Pham Alexander , Tisch Roland 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune disease mediated by pathogenic beta cell-specific T cells. The use of antigen-specific based immunotherapies has been one strategy to selectively target beta cell-specific T cells leaving the remainder of the immune system intact. Administration with peptides derived from beta cell autoantigens such as glutamic acid decarboxylase 65 (GAD65) to NOD mice prevents T1D. However, as beta cell autoimmunity progresses the conditions for peptide treatment to suppress T1D become highly stringent. To enhance the efficacy of peptide treatment, recombinant IAg7-immunoglobulin (Ig) dimers covalently linked to GAD65 peptides (217–236, 286–309) or the mimetic BDC2.5 epitope (sBDC) were tested. Twelve week-old NOD female mice with established beta cell autoimmunity received a short course of IAg7-Ig dimers intravenously. Surprisingly, treatment with IAg7-Ig:sBDC accelerated the onset of diabetes in NOD mice due to expansion of sBDC-specific CD4+ Th1 cells. In marked contrast, the majority of NOD mice treated with IAg7-Ig:GADp217 or IAg7-Ig:GADp286 remained diabetes-free. Protection correlated with an increased frequency of IL-10 secreting immunoregulatory CD4+ T cells that blocked diabetes in a co-adoptive transfer model. These results demonstrate that treatment with a short-course of IAg7-Ig-GAD65 peptide dimers is an effective approach to suppress T1D.