bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mice immunized with EBNA-1 protein produce antibodies that cross-react with double stranded DNA (129.5)
Authors: Spatz Linda Ann , Tran Hoa , Mumbey-Wafula Alice , Chandrasekhar Sharanya , Wei Hoi , Lewis Rita , Gottlieb Paul 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) is characterized by the production of antibodies to nuclear antigens, in particular antibodies to double stranded DNA (dsDNA). The etiology of these autoantibodies is unknown but one theory suggests that viruses may play a role. One virus that has been shown to have an association with SLE is the Epstein Barr Virus (EBV). We previously demonstrated that mice immunized with an EBNA-1 expression vector or with purified EBNA-1 protein develop antibodies to dsDNA as well as to the viral antigen. We have recently demonstrated that these antibodies are pathogenic and deposit in the kidney and induce moderate proteinuria. We hypothesized that since EBNA-1 is a dsDNA binding protein, it could complex with chromosomal dsDNA in order to render it immunogenic and elicit an anti-dsDNA response. To test this hypothesis we generated recombinant EBNA-1 protein that lacks the 3 known chromosomal binding sites and immunized mice with it. We observed that mice immunized with this mutated EBNA-1 protein still develop an anti-dsDNA response. We have now generated hybridomas producing anti-EBNA-1 antibodies from these mice and observed that 10 out of 18 monoclonal anti-EBNA-1 antibodies actually cross-react with dsDNA. These results suggest that a viral protein determinant can mimic chromatin and elicit an anti-dsDNA response that can lead to autoimmunity. Future studies will analyze the molecular basis of this cross-reactivity.
The CYP450 mouse model for autoimmune hepatitis: Breaking of self-tolerance in transgenic CYP2D6 and wildtype FVB-mice by viral infection (130.1)
Authors: Christen Urs , Holdener Martin , Hintermann Edith , Johnson Eric , Manns Michael , von Herrath Matthias 
Published: January 2023

Abstract
The etiology of autoimmune hepatitis (AIH) is poorly understood although the major autoantigen, cytochrome P450 2D6 (CYP2D6), has been identified. We generated an animal model for human AIH using the natural autoantigen CYP2D6. We infected transgenic mice expressing human CYP2D6 in the liver with an Adenovirus-CYP2D6 vector (Ad-2D6) to break self-tolerance. Upon infection with Ad-2D6 not only transgenic CYP2D6 mice but also wildtype FVB mice showed persistent features of severe liver damage associated with AIH (hepatic fibrosis, fused liver lobules, cellular infiltrations, elevated serum ALT levels, confluent necrosis). Ad-2D6-infected mice (CYP2D6 and FVB) generated high titers of anti-CYP2D6 antibodies. Epitope mapping revealed that anti-CYP2D6 antibodies predominantly recognized the same immunodominant linear epitope recognized by sera of AIH patients (AQPPRD aa265-270). In contrast, mice infected with a control Adenovirus expressing green fluorescence protein did neither develop liver damage nor generated anti-CYP2D6 antibodies. The severity of liver damage as well as antibody formation was enhanced in FVB mice compared to transgenic CYP2D6 mice indicating a stronger tolerance to human CYP2D6 in CYP2D6 mice. In FVB mice, due to the homology of the mouse isoenzymes of the CYP superfamily to human CYP2D6, autoimmune liver damage by Ad-2D6-infection was possibly induced via true molecular mimicry.
CD73-deficient mice are resistant to experimental autoimmune encephalomyelitis (130.30)
Authors: Bynoe Margaret S. , Mills Jeffrey , Thompson Linda F 
Published: January 2023

Abstract
To test the role of CD73-generated adenosine in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, we induced EAE in CD73−/− mice with standard protocols. CD73 (ecto-5’-nucleotidase) makes extracellular adenosine from AMP. Adenosine is anti-inflammatory in several settings via triggering cell surface adenosine receptors and CD73−/− mice exhibit a variety of pro-inflammatory phenotypes. However, CD73−/− mice were highly resistant to the development of EAE (mean EAE score of 0.5 vs 2.0 for WT mice). This was not because their T cells failed to be activated by EAE induction. When T cells from CD73−/− and WT mice with EAE were transferred to CD73+/+ TCRα−/− mice, disease activity was much more severe in recipients of CD73−/− T cells (mean EAE score 3.7 vs. 1.7 in WT mice). To determine whether CD73−/− T cells were defective in their ability to cross the blood brain barrier, immunohistochemistry was performed on the brains of CD73+/+ and CD73−/− mice with EAE. Consistent with the lower disease activity scores in the CD73−/− mice, there were many fewer leukocytes in their brains. The higher apparent blood brain barrier function in CD73−/− mice was confirmed with Evan’s blue dye extravasation studies. Thus, our results show that CD73is required for the entry of pathogenic T cells into the brain and suggest a novel role for CD73-generated adenosine in modulation of the blood brain barrier.
Induction of CD8 T cell-mediated EAE by viral infections in a CD8 TCR transgenic mouse model (130.31)
Authors: Ji Qingyong , Perchellet Antoine , Goverman Joan 
Published: January 2023

Abstract
Experimental autoimmune encephalomyelitis (EAE) is an animal model for multiple sclerosis (MS), an inflammatory demyelinating disease of the central nervous system (CNS). Besides CD4 T cells, CD8 T cells have recently been shown to mediate the pathogenesis of EAE. The cause of MS is not clear, but infections have been suggested. In this study, we investigated the roles of viral infections in the development of EAE on a CD8 T cell receptor (TCR) transgenic mouse model. Infection with wildtype vaccinia virus broke tolerance and triggered atypical EAE in a TCR transgenic mouse specific for myelin basic protein 79–87 (MBP79-87). CD8 T cells up-regulated activation markers and trafficked to the CNS in virus-infected mice. Interestingly, CD8 T cells from TCR-transgenic MBP deficient mice also displayed activation markers, and had cytolytic effector function similar to that from TCR transgenic MBP sufficient mice. T cell activation was not due to MBP TCR cross-reactivity against vaccinia virus. Furthermore, preliminary data showed that disease induction by vaccinia virus required endogenous TCRs. Ongoing experiments are extending these observations into another viral infection. In conclusion, the results suggest that viral infection may induce EAE by activation through dual TCR.
CD8 epitope MOG 37-46 induces attenuated form of EAE characterized by reduced CNS infiltration and myeloid activation (130.32)
Authors: Bettini Maria , Evavold Brian D 
Published: January 2023

Abstract
Multiple sclerosis is a predominantly CD4 T cell mediated disease, yet recent observations have indicated a potential contribution from CD8 T cells. We have previously identified MOG 37-46 in the context of H2-Db as the CD8 specific epitope and showed that most CD8 T cells in the CNS of MOG 35-55 immunized mice are specific to MOG 37-46. In an attempt to isolate the contribution of CD8 T cells in a murine model of MS, we immunized mice with MOG 37-46, a truncated version of MOG 35-55. MOG 37-46 does not contain the full T helper epitope (40–48) and shows minimal activation of CD4 T cells. The data presented here show that MOG 37-46 immunization induced disease characterized by lower disease scores, a decrease in CNS infiltration and microglial activation, and reduced demyelination and axon damage. Upon re-stimulation splenic-derived CD8 T cell lines from MOG 35-55 and MOG 37-46 immunized mice were IFNγ+ and TNFα+ (&gt;60%). CNS derived CD8 T cells of EAE mice produced IFNγ and TNFα in response to antigen after ex vivo re-stimulation. Our data indicate that CD8 T cells reactive to MOG 37-46 are pro-inflammatory, and suggest that CD4 T cells are required for myeloid activation in CNS and recruitment of CD8 T cells.
This research was supported by NIAID (R01 AI056017) and NMSS (RG 3683) grants.
Cyclophosphamide treatment of the NOD mouse reveals quasi-public dominant T cell clones (130.34)
Authors: Marrero Idania G , Dai Yang D , Sercarz Eli E 
Published: January 2023

Abstract
To define the major components of the autoreactive T cell repertoire that arise spontaneously in the NOD mouse, we used CDR3 length spectroscopy to
study the T cell repertoire in PLN from pre-diabetic NOD mice to identify public dominant clones; andto identify potentially pathogenic clones in islet-infiltrating cells that persist after Cyclophosphamide (Cyp) treatment.
We found that any public T cell expansions were identified in PLN of female NOD mice at 4 and 10 weeks of age. Two expansions in the rearrangements Vb5.2/Jb2.5 and Vb10/Jb1.5 were found in 60% and 50% of 4 week old NOD mice. These expansions can be useful in the study of how the T cell repertoire can be modified after islet transplantation. After Cyp-treatment, the Vb4, Vb5.2, Vb10 and Vb12 families showed dominant expansions. Interestingly, we found a unique Vb4-"195 nucleotide" peak that was found in 10 out of 11 (91%) Cyp-treated mice. In contrast, this peak was not detected in untreated NOD mouse islets. In the Vb12 family, a "200" peak was prominent in 6 out of 10 (60%) Cyp-treated mice. These expansions may represent the more pathogenic clones. Currently, we are sequencing these expansions to establish whether the Cyp-induced expansions represent truly unique clones. We believe that these Cyp experiments will provide the possibility of distinguishing between regulatory and aggressive clones.
This work was supported by JDRF 3-2005-336.
T cell receptors and I-Ad/peptide interactions in autoimmune gastritis (AIG) (130.40)
Authors: Revilleza Maria Jamela Roxas , Levin Ditza , Mage Michael , Teyton Luc , Shevach Ethan , Margulies David H , Natarajan Kannan 
Published: January 2023

Abstract
We have developed a murine model of autoimmune gastritis by generating two TCR transgenic lines expressing I-Ad restricted TCRs specific for two different peptide epitopes of gastric H/K ATPase. Both lines spontaneously develop AIG but with differing incidence, severity, and kinetics. The Tx23 line develops disease early, severely, with 100% incidence and a predominantly Th1 phenotype. The Tx51 line develops disease only after two months, only 50–80% of the mice are affected. and is predominantly of the Th2 type. To investigate the role of TCR affinity in the disease phenotypes in these mice, we generated recombinant, soluble forms of the Tx23 and Tx51 TCRs, and monomeric and multimeric forms of their cognate I-Ad/peptide complexes. Peptide dose response experiments suggest that the Tx51 TCR is of higher functional avidity than the Tx23 TCR, suggesting that decreased peptide availability in vivo may limit the activation of Tx51 T cells. Surface plasmon resonance (SPR) studies revealed specific and dose dependent binding when multimeric but not monomeric I-Ad/peptide complexes were offered to the TCR, consistent with the relatively weak affinities of these autoreactive TCRs. These experiments exploit strategies that may be employed to detect binding of low affinity TCRs to their MHC/peptide ligands.
This was supported by the Intramural Research Program of the NIAID.
Serum transfer of spontaneous polychondritis from HLA-DQ6αβ 8αβ transgenic mice. (130.42)
Authors: Doll Joseph R , Bradley David S 
Published: January 2023

Abstract
We provide here evidence for a pathogenic role for antibodies in spontaneous polychondritis occurring in HLA-DQ6αβ8αβ transgenic (tg) mice. We have previously reported that HLA-DQ6αβ8αβ tg mice develop spontaneous polychondritis, which occurs at middle age and lacks CII specificity. The onset of disease is associated with elevated serum levels of immunoglobulins compared to non-diseased mice. However, a role for immunoglobulins in disease pathogenesis has not been identified. Serum was collected from HLA-DQ6αβ8αβ tg mice displaying clinical spontaneous polychondritis and serum immunoglobulins were concentrated via density centrifugation. Concentrated serum immunoglobulins were transferred intravenously into young, naïve B10.T(6R) mice, resulting in a transient clinical display of polyarthritis. Gross clinical symptoms of polyarthritis occurred within 48 hours of the initial immunoglobulin transfer and were of a transient nature, symptoms generally reduced by 96 hours. Histological assessment of the limbs obtained from mice receiving antibody transfer identified the presence of cellular infiltrates in the synovium and peri-synovium. Control mice receiving sham immunizations displayed neither clinical nor histological signs of polyarthritis. The studies described here should aid in characterizing and understanding a newly identified transgenic mouse model of human autoimmune polychondritis and may provide insight into new therapeutic avenues for human relapsing polychondritis and related diseases such as rheumatoid arthritis. In addition, these findings may aid in the identification of unidentified target antigens required for spontaneous polychondritis.
DRB1*0402 protects while DRB1*0401 is permissive for inducing collagen-induced arthritis in transgenic mice (130.45)
Authors: Taneja Veena , Behrens Marshall , Griffiths Marie , Luthra Harvinder , David Chella 
Published: January 2023

Abstract
DRB1*0401 has been associated with predisposition to rheumatoid arthritis while DRB1*0402 is associated with resistance. We have generated transgenic mice lacking endogenous H2-A and E molecules (AEo) but expressing DRB1*0401 or DRB1*0402 molecules. All transgenic mice expressed functional DR molecules which could positively and negatively select various Vβ T cell repertoire profile. Transgenic mice were studied in vivo for induction of arthritis following immunization with type II collagen. DRB1*0401 mice are susceptible to collagen-induced arthritis while DRB1*0402 mice are resistant. Presence of DR4 leads to arthritis predominantly in female mice. CIA susceptible AEo.DQ8 mice have been shown to develop arthritis without a gender bias. However, AEo mice expressing DRB1*0401.DQ8 develop arthritis in female mice with a ratio of 3F:1M, similar to that in humans. DRB1*0402 mice did not show any defect in humoral response though they demonstrated a lower cellular response to CII and an increased activation-induced cell death compared to susceptible mice. In vitro T cell response with superantigen staphylococcal enterotoxin B and Concovalin A showed that DRB1*0402 mice are not defective in generating T cell response. The resistant mice showed an increased cell death in thymus suggesting deletion of autoreactive T cells at thymic level might be one reason for protection. The data suggest that DRB1 polymorphism lead to selection of TCR in thymus thus controlling the selection of autoreactive T cells available in periphery. Thus these transgenic mice may be useful to study the reasons for gender-bias observed in human patients develop therapeutic strategies.
Microsomal prostaglandin E synthase-1 genetic deletion results in reduced incidence and severity of collagen-induced arthritis associated with a marked reduction of anti-collagen antibodies (130.47)
Authors: Kojima Fumiaki , Kapoor Mohit , Yang Lihua , Crofford Leslie J 
Published: January 2023

Abstract
Microsomal prostaglandin E synthase-1 (mPGES-1) is an inducible enzyme that acts downstream of cyclooxygenase and specifically catalyzes the conversion of prostaglandin (PG)H2 to PGE2, most prominently in inflammatory conditions. However, the role of mPGES-1 in the immune response has not yet been full elucidated. In this study, we demonstrate the effect of genetic deletion of mPGES-1 on initiating immune responses as well as clinical arthritis development in bovine collagen-induced arthritis (CIA), a well established model to study pathogenic mechanisms relevant to rheumatoid arthritis. mPGES-1 null and heterozygous mice exhibited decreased paw edema, arthritis severity and incidence compared to wild-type mice in a gene dose-dependent manner. Histopathological changes further showed a significant reduction in lining hyperplasia and tissue destruction in mPGES-1 null mice compared to their wild-type littermates. In addition, mPGES-1 genetic deletion showed attenuated mechanical allodynia in a gene dose-dependent manner. The reduction of inflammatory features observed in mPGES-1 null mice were associated with significantly lower levels of IgG including IgG1, IgG2a, IgG2b, IgG2c and IgG3 against type II collagen (CII). These results suggest that deficiency of mPGES-1 inhibits the development of CIA, at least in part, by blocking development of the anti-CII antibody response.
Grant Support: Arthritis Foundation Biomedical Sciences Grant and NIH/NIAMS AR49010.
OX40 blockade delays disease progression in NOD by inducing qualitative changes in Tregs (131.1)
Authors: Honkanen-Scott Minna Maria , Nettles Adam , Bansal-Pakala Pratima 
Published: January 2023

Abstract
Costimulatory interaction between T cells and APCs is crucial for eliciting an immune response. By blocking costimulatory pathways it is possible to prevent or slow disease progression in many autoimmune diseases. We previously investigated the role of OX40/OX40L blockade in T1D. We demonstrated that OX40 blockade suppressed T1D progression when given at late stage but before T1D symptoms. These data are consistent with the role of OX40 as secondary costimulatory molecule. In the present study we investigated the mechanisms by which OX40 blockade suppresses T1D progression in NOD.
OX40 blockade marginally reduces expression of CD80 on DCs. There is a reduction in frequency of islet-reactive effector T cells in spleen and LN. Migration of effector T cells to pancreas may be inhibited since greatly reduced lymphocytic infiltrate was observed in the pancreas. Interestingly, OX40 blockade did not enhance Treg numbers in spleen, draining LN or the pancreas. We did not observe a reduction in Tregs in control animals, contrary to previous reports. In addition, FoxP3+ cells were present in and around infiltrated islets, suggesting that the number or location of Tregs may not be defective in NOD. Preliminary data suggests that OX40 blockade induces qualitative changes in Tregs by changing the regulatory mechanisms. Our data suggests that changes in function, rather than number, of Tregs may be crucial for suppressing T1D.
ARALAST or ANTI-TNF-a TREATMENT PREVENTS AUTOIMMUNE DESTRUCTION OF ISLET GRAFTS IN NONOBESE DIABETIC (NOD) MICE (131.23)
Authors: Koulmanda Maria , Hoffman Lauren , Qipo Andi , Shi Hang , Fan Zhigang , Flier Jeffrey F. , Strom Terry B. 
Published: January 2023

Abstract
The onset hyperglycemia in NOD mice coincides with loss of a critical portion of the islet cell mass. Thus, restoration of a portion of the lost islet cell mass should be sufficient to create euglycemia if a deficit in insulin producing tissue is the sole cause of disease. It is notable that many potent T-cell tolerizing therapies have failed to enable restoration of euglycemia in new onset T1D NOD mice with or without transplantation of islets. Hence, we have searched for other potentially complicating features, e.g. insulin resistance, that may serve as an additional barrier to restoration of euglycemia. We now report that new onset T1D is associated with inflammation induced insulin resistance created by faulty phosphorylation of crucial elements of the insulin signaling pathway (insulin receptor and IRS-1). To test the hypotheses that inflammation, insulin resistance and T1DM disease expression are linked we have tested the effects of select anti-inflammatory agents in T1DM NOD mice. We now report that treatment for 15 days with alpha-1-antitrypsin (AAT) or 20 days with anti-TNF-a (i) permanently restores a euglycemic state in 14 of 16 or 22 of 24 treated new onset diabetic NOD mice (ii) ablated insulin resistance and (iii) produces specific tolerance to syngeneic, not acceptance of allogeneic, islets that is manifest long after cessation of treatment of new onset diabetic NODs while preserving other immune responses. In short, the restoration of euglycemia, self-tolerance, and of the faulty insulin receptors and IRS- 1 phosphorylation patterns can be corrected by treatment with ATT, an acute phase reactant with potent anti-inflammatory activity, and/or anti-TNF-a.
CD4+CD25+Foxp3+ T regulatory cell stimulation by high dose gp96 is costimulation and APC independent (131.3)
Authors: Nevius Erin , Binder Robert , Srivastava Pramod 
Published: January 2023

AbstractThe immunogenicity of heat shock protein-peptide complexes (HSPs) against tumors and pathogens is initiated when the HSP engages its receptors, CD91 and maybe others, on antigen presenting cells (APCs). This allows APCs to obtain antigen in the form of peptides, and acquire signals that indicate for them to mature, express co-stimulation and migrate (1). However, high doses of HSPs tend to negate ongoing T cell responses (2), an effect shown to involve generation of a CD4+ suppressor response (3). This immune suppression has been demonstrated to protect mice from autoimmune conditions including experimental autoimmune encephalomyelitis (EAE) and the NOD model of diabetes (3). Here we provide evidence that highly enriched CD4+CD25+Foxp3+ T regulatory cells proliferate up to 3 fold more in the presence of the HSP gp96 and IL-2 compared to IL-2 alone. LPS or albumin were used as controls and did not support proliferation. This effect is APC and co-stimulation independent and may occur via CD91 which is expressed on T regulatory cells or by another receptor. Overall CD4+CD25+Foxp3+ T regulatory cells are stimulated directly by HSPs and may be the mechanism by which high doses of HSPs induce a suppressor response.This study was supported by NIH grant CA84479-02 and by a research agreement with Antigenics Inc.
Application of humanized FcRn mouse model for in vivo testing of therapeutic antibodies (131.37)
Authors: Petkova Stefka B , Meng Yu-Ju Gloria , Sproule Tomas J , Christianson Gregory J , Roopenian Derry C 
Published: January 2023

Abstract
IgG antibodies modified for high binding affinity for their receptor FcRn have emerged as a major treatment for a wide array of both neoplastic and autoimmune diseases. The goal of this study was to analyze the effect of human IgG1 antibodies with enhanced in vitro binding to FcRn on their in vivo half-life in mice deficient in mouse FcRn and expressing human FcRn. Two mutants (N434A and T307A/E380A/N434A) of the humanized monoclonal antibody Hu4D5, directed against epidermal growth factor receptor 2 (p185 HER2), show increased pH-dependent binding to human FcRn in vitro. They also have an extended half-life in vivo compared to wild-type antibody in FcRn transgenic mice but not in mice expressing endogenous mouse FcRn. When injected into FcRn-humanized mice at a concentration sufficient to partially saturate human FcRn, the mutants with an extended serum half-life were most effective in reducing the half-life of a tracer antibody. Finally, we use a serum-transfer model of arthritis and FcRn-humanized mice to test whether mutant antibodies could have enhanced therapeutic potential by promoting accelerated clearance of arthritogenic antibodies. Results indicate that the engineered mutant T307A/E380A/N434A is most effective in vivo in ameliorating the inflammatory arthritic lesions.
In summary, FcRn-humanized mice are a promising animal model for human IgG therapeutic development.