bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel approach to generating disease modifying dendritic cells (DCs) (129.13)
Authors: Papenfuss Tracey L. , Kithcart Aaron P. , Powell Nicole D. , McClain Melanie A. , Gienapp Ingrid E. , Shawler Todd M. , Whitacre Caroline C.
Published: January 2023

Dendritic cells (DCs) bridge the innate and adaptive immune responses and DC immunotherapy has shown promise in the treatment of transplantation, allergic and autoimmune diseases. One primary limitation in studying and utilizing DCs is their small numbers in vivo. While the growth factor Flt3-ligand (FL) is used to expand DCs in vivo, GM-CSF is predominantly used to generate DCs in vitro from bone marrow (BM-DC). The purpose of this study was to explore the use of murine FL (mFL), the mouse homologue of human FL, to generate DCs in vitro. In vivo (tumor-based) mFL delivery reliably expanded splenic DCs comparable to hFL In vitro BM-DCs generated with mFL were more similar to hFL than GM-CSF derived BM-DCs and appeared to be immature. Specifically, CD11c, CD11b and maturation markers CD80 and CD86 were lower in mFL and hFL versus GM-CSF BM-DCs. Maturation of mFL BM-DCs with TNF-α±LPS increased CD80, CD86 and CD40 expression. Transfer of matured mFL BM-DCs to mice prior to immunization with myelin oligodendrocyte glycoprotein (MOG35-55) significantly increased cumulative disease (26.79±10.12) and peak disease scores (3.07±1.16) compared to immature mFL BM-DCs (20.21 ±7.64 and 1.86±0.7, respectively). These results suggest that exposure to mFL in vitro is a novel method to generate large numbers of immature BM-DCs which can modulate the course of autoimmune disease.

(Support: NIH AI-43376, K01 R022198 and NMSS RG3272).
Human DcR3 overexpression in mice results in a systemic lupus erythematosus-like syndrome (129.15)
Authors: Wu Jiangping , Han Bing , Moore Paul , Luo Hongyu
Published: January 2023

OBJECTIVE: DcR3, a TNFR family member, is a secreted protein, and can enhance cell survival by interfering with multiply apoptosis pathways. The objective of this study was to understand the role of DcR3 in pathogenesis of autoimmune diseases.

METHODS: We generated transgenic (Tg) mice with actin promoter-driven expression of human DcR3, and investigated the development of autoimmune diseases in these mice.

RESULTS: Beyond 5–6 months of age, DcR3 Tg mice developed a systemic lupus erythematosus (SLE)-like syndrome. They produced autoantibodies against various organs, and against dsDNA and Smith Ag. The kidneys of these Tg mice showed pathological changes indicative of glomerular nephritis and IgG and C3 deposition; kidney dysfunctions, such as proteuria, leukocyturia, and hemuresis, were obvious. Aged Tg mice also developed skin lesions and lymphocyte infiltration in the liver, and suffered from leukopenia, anemia and thrombocytopenia. The SLE-like symdrome penetrance in DcR3 Tg mice was gender-dependent, with about 60% in females versus 20% in males. Mechanistically, exogenous recombinant DcR3 or endogenous DcR3 produced by Tg T cells effectively protected CD4 cells from activation-induced apoptosis in vitro. Probably as a consequence, in the peripheral blood of Tg mice beyond 6 months of age, CD4 cells with a phenotype of previous activation were increased.

CONCLUSION: DcR3 overexpression could lead to a SLE-like syndrome in mice.
Expression of transgenic FLIP on thyroid epithelial cells in CBA/J mice inhibits induction and promotes resolution of granulomatous experimental autoimmune thyroiditis (G-EAT) (129.17)
Authors: fang yujiang , Braley-Mullen helen
Published: January 2023

G-EAT is induced by transfer of thyroglobulin-primed in vitro activated splenocytes. In EAT susceptible DBA/1 and CBA/J mice, inflammation either resolves or progresses to fibrosis by day 60 depending on the extent of damage at day 20. Depletion of CD8+ T cells inhibits G-EAT resolution. We previously showed that FLIP transgene (Tg) expressed on thyroid epithelial cells (TEC) of DBA/1 mice had no effect on induction, but promoted earlier resolution of G-EAT. However, when Tg+ CBA/J mice expressing FLIP on TEC were used as recipients of WT donor splenocytes, they developed less severe G-EAT than FLIP Tg− littermates (average severity score of 1.3 vs. 4.5 at day 20), although both strains had similar levels of FLIP transgene. Following transient depletion of CD8+ T cells, FLIP Tg+ and Tg− CBA/J recipients both developed severe G-EAT (severity score of 4.4) at day 20. Lesions in CD8-depleted Tg+ recipients were resolving by day 60, whereas lesions in Tg− littermates did not resolve and most were fibrotic. FLIP and FasL were mainly expressed by TEC in FLIP Tg+ recipients and by inflammatory cells in Tg− recipients. mRNA expression of IL-10 and IL-13 was higher, and IFN-γ and TNF-α were lower, in FLIP Tg+ compared to Tg− recipients. The results support the hypothesis that transgenic FLIP expressed on TEC in CBA/J mice promotes G-EAT resolution, but induction of G-EAT is inhibited unless CD8+ T cells are depleted (NIH Grant DK35527).
IFN-gamma inhibits inflammatory arthritis via suppression of mast cell activation. (129.23)
Authors: Miyake Sachiko , Kaieda Shinjiro , Oki Shinji , Yamamura Takashi
Published: January 2023

KBxN mice spontaneously develop arthritis similar to RA in humans, and transfer of serum from arthritic K/BxN mice into healthy animals provokes arthritis within days independently on T and B cells. K/BxN serum-induced arthritis is mediated by anti-glucose-6-phsophate-isomerase IgGs. Here we demonstrate that synthetic glycolipid ligand, a-galactocylceramide (alpha-GalCer) or its analogue with an elongated sphingosine chain strongly suppressed K/BxN serum transfer arthritis by inhibiting inflammatory cellular infiltration and subsequent destruction of cartilage and bone. The inhibitory effect mediated by these glycolipd ligands was abolished by neutralization of IFN-gamma and systemic administration of IFN-gamma prevented the development of inflammatory arthritis. Furthermore, we demonstrate that histamine release involved in the induction of arthritis was suppressed by administration of glycolipid ligands or IFN-gamma. In addition, degranulation of mast cells in synomium was significantly suppressed in glycolipid ligands-treated mice, suggesting that the suppression of mast cell activation contributes to the inhibition of arthritis.
Th17 Cells Alone are not Sufficient to Induce CNS Autoimmunity, but can Synergize with Th1 Cells to Induce EAE (129.24)
Authors: Shriver Leah , Mann Monica , Dittel Bonnie N
Published: January 2023

During a primary immune response, CD4 T cells can polarize into two subsets, Th1 or Th2 depending on the cytokine environment. These subsets have been differentially implicated in the pathogenesis of autoimmune (Th1) or allergic (Th2) disorders. Recently a new lineage of T helper cells has been described, Th17 cells, which produce the cytokine IL-17, and evidence has pointed to these cells as key mediators of autoimmune disorders such as the inflammatory demyelinating disease multiple sclerosis (MS). A novel in vitro skewing paradigm shows that the cytokines IL-6, IL-1, TGF-β, and IL-23 are responsible for the development of this lineage. However, to date, no study has examined whether these cells are solely responsible for induction of CNS autoimmunity. We examined this question using an adoptive transfer model of MS in B10.PL mice termed experimental autoimmune encephalomyelitis (EAE). We found that 1 x 106 MBP-specific Th1 cells induced a monophasic disease course, while the same number of Th17 cells did not induce EAE. These data suggest that Th17 cells alone are not sufficient to initiate EAE disease. The adoptive transfer of 0.125 x 106 Th1 T cells did not result in EAE, but when mixed with 1 x 106 Th17 cells the mice exhibited full blown EAE. In a coculture model of immune cell-neuron interaction that mimics neuronal dysfunction during EAE, we found that Th1 cells induced neuronal dysfunction, while Th17 cells did not. These cumulative data suggest that the two T cell subtypes play differential roles that are synergistic during CNS inflammation and that therapeutic targeting of Th17 cells alone may not prevent pathology in diseases such as MS.
Signal via IL-12Rβ2 plays an essential role in the development of CD4+CD25+ regulatory T cells (129.27)
Authors: Zhao Zhao , Zhang Guang-Xian , Yu Shuo , Ciric Bogoljub , Fitzgerald Denise , Rostami A.M.
Published: January 2023

It has been recently found that mice lacking IL-12 specific receptor subunit, namely IL-12Rβ2, developed more severe clinical and pathological signs of in experimental autoimmune encephalomyelitis (EAE), an animal model of human multiple sclerosis. However, whether this phenomenon is also true in other autoimmune disorders and the mechanism underlying this phenomenon is not known. We in the present study have shown that IL-12Rβ2-deficient mice also develop earlier and more severe disease in the streptozotocin-induced model of diabetes, indicating a universal susceptibility to autoimmune diseases in IL-12Rβ2-deficient mice. IL-12Rβ2-deficient mice exhibited significantly higher effector T cell proliferative responses not only to autoantigens, but also to non-specific stimuli. Compared to wild type mice, IL-12Rβ2-deficient mice exhibited significantly reduced capacity to develop regulatory T cells (Treg) in vivo in the thymus and in vitro upon stimulation with TGF-β. Further, naturally-occurring Treg cells from these mice were less functional in suppressing effector T cells. These findings indicated that signal via IL-12Rβ2 plays an essential in developing high quantity and quality of Treg cells, which confers a novel mechanism underlying the regulation of autoimmune diseases by the IL-12 pathway.
Monoclonal B Cell Expansion and Antigen-driven Somatic Hypermutation of Ig VH Genes in Synovial Membrane of Patients with Crystal-Induced Arthritis (129.3)
Authors: Kolln Johanna , Da Reng-Rong , Qin Yufen , Baeten Dominique , Zhang Yiping
Published: January 2023

The mechanisms underlie the inflammation in crystal-induced arthropathies (CyA) is not fully understood. Inflammatory mediators, like crystal-specific antibodies and macrophages have been explored as possible factors in the generation or progression of CyA. To examine whether B cells are involved in the pathogenesis of CyA, we analyzed immunoglobulin (Ig) heavy chain variable region (VH) genes of B cells from synovial membrane of five CyA patients. Using a RT-PCR technique, we could detect rearrangements of Ig VH genes in all five cases. Sequence analysis of CDR3 region of rearranged VDJ genes revealed a monoclonal B cell expansion in four out of five CyA patients. The nucleotide sequences of VH genes from clonally expanded B cells demonstrated the heterogeneous usage of the Ig VH gene family. All expanded B cell clones showed somatic mutations in the CDRs with a high replacement-to-silent ratio (> 2.9) and were distributed in a way suggesting that these B cells had been positively selected through their antigen receptor. The intraclonal variations is also common, indicate that consistent antigen stimulation is close related to CyA development. These data demonstrate that there are antigen-driven clonal accumulations of B cells in CyA synovial membrane. Whether these antibodies are specific to crystal epitopes is currently under investigation.

This study was supported by the NIH Grant RO1 AI 54911.
IL-4 promotes experimental drug-induced hepatitis by reducing immune responses to haptenated proteins and inhibiting regulatory responses to native CYP2E1 (129.38)
Authors: Njoku Dolores B. , Li Zhaoxia , Talor Monica V , Rose Noel R.
Published: January 2023

Susceptible persons develop drug-induced hepatitis (DIH) following administration of halogenated anesthetics or similar drugs. IL-4 worsens autoimmune hepatitis by promoting Th2 responses while other possible mechanisms for this observation have not been fully investigated. Using our experimental model of anesthetic DIH created by immunizing mice with hepatic proteins covalently linked to the TFA hapten (TFA-S100) formed from CYP2E1 metabolism of anesthetics, we found that IL-4 deficient (KO) mice developed significantly less hepatitis than BALB/c (WT) mice. Moreover, ex vivo, antigen specific T cells to CYP2E1 and TFA from IL-4 KO mice proliferated significantly less than those from WT mice. To uncover additional IL-4-dependent mechanisms for the development of anesthetic DIH we measured supernatant cytokines from splenocytes stimulated in vitro with TFA or CYP2E1 and obtained from WT and KO mice following TFA-S100 immunization. KO mice had less IL-1β (p < 0.01), IL-2, IFN-γ, TNF-α, (p < 0.05), IL-13 (p < 0.05) and IL-6 (p < 0.05) in response to TFA when compared to WT mice, suggesting that IL-4 had a critical role in the development of immune responses to TFA-haptenated proteins. Ex vivo, antigen specific T cells to CYP2E1 had higher levels of IL-10 (p < 0.01) suggesting that IL-4 promotes anesthetic DIH by diminishing regulatory responses to CYP2E1.

Supported by R21DK075828.
Characterization of immune response in â-tubulin induced murine experimental autoimmune hearing loss (129.44)
Authors: Zhou Bin , Glickstein Jonathan , Zhou Yixuan , Kermany Mohammad Habiby , Cai Chun , Cai Qing , Kim Jun Woo , Kim Patrick , Liu Wenxia , Yoo Tai June
Published: January 2023

The pathological mechanism(s) of autoimmune sensorineural hearing loss (ASNHL) is still unknown. An autoimmune reaction to inner ear structures could be one of several possible pathogenic factors involved in ASNHL. In the current study we show that the inner ear–specific proteins b-tubulin are capable of targeting experimental autoimmune hearing loss (EAHL) in mice. Two weeks after immunization of BALB/c mice with b-tubulin, auditory brainstem responses (ABR) showed significant hearing loss, and the hearing loss can lasted at least six weeks. The antibodies activity to â-tubulin increased in dose dependent compared with controls. In addition, sera of these mice contained high levels of anti-â-tubulin antibodies could transferred EAHL to naive mice. Flow cytometry analysis showed that b-tubulin activated CD4+ T cells with a proinflammatory Th1-like phenotype. T cell mediation of EAHL was determined by showing significantly increased ABR thresholds 6 weeks after adoptive transfer of b-tubulin-activated CD4+ T cells into naive BALB/c recipients. Immunocytochemical analysis showed that leukocytic infiltration of inner ear tissues, and histological analysis showed that the loss of inner and outer hair cells in â-tubulin-immunized mice. Moreover, the percentage of CD4+CD25+ regulatory T cells and the level of FoxP3 were significantly decreased in EAHL. Our study provides an experimental confirmation that ASNHL may be a humoral and cellular immune reactivity against b-tubulin, which may be sufficient for mediating the hearing loss observed in ASNHL.
Increased Amyloid β Protein Levels in Children with Down Syndrome (129.47)
Authors: Mehta Pankaj D , Mehta Sangita P , Capone George
Published: January 2023

Objective: To quantitate plasma amyloid β protein (Aβ)40, Aβ42, and neopterin levels in Down Syndrome (DS) children and controls.

Background: DS persons (>40 years) have neuropathological changes characteristic of Alzheimer disease (AD). Soluble forms of Aβ generated from amyloid precursor protein (APP) end at C-terminal residues 40 and 42. Neopterin is an immune activation marker for the cell-mediated immune response.

Design/Methods: Blood was collected from DS (N = 35; 7 ± 3.8 years old) and their siblings (N = 34; 10 ± 4.5). Plasma Aβ40, Aβ42, and neopterin levels were quantitated by ELISA.

Results: Aβ40 (Mean ± SD; 277 ± 70 pg/ml) and Aβ42 (31 ± 8 pg/ml) levels were higher in DS children than controls (Aβ40 155 ± 35 pg/ml, Aβ42 22 ± 5 pg/ml) (p<.0001). There were significant negative correlations between age and Aβ40 in DS (r=.47, p<.004) or controls (r=.38, p<.026) and Aβ42 levels in DS (r=.61, p<.001) but not in controls. Neopterin levels were higher in DS (3.01 ± 1.37 ng/ml) than controls (1.57 ± 0.65 ng/ml) (p<.0001). There was no correlation between neopterin levels and age, and Aβ40 or Aβ42 levels in DS or controls.

Conclusion: The overexpression of APP gene in DS leads to increases in plasma Aβ40 and Aβ42 levels before plaque formation in DS brain. A lack of correlation between neopterin and Aβ levels suggests that higher neopterin concentrations in DS reflect inflammatory cell activation rather than AD neuropathology.
Cloning and characterization of nucleolar-specific autoantibodies from a murine model of lupus/scleroderma (129.6)
Authors: Kole Hemanta K , Bolland Silvia
Published: January 2023

FcγRIIB knock-out mice in B6 background develop a lupus like autoimmune disease. With the addition of the yaa (Y-chromosome autoimmune accelerator) gene, pathogenecity in these male mice is significantly enhanced and autoantibodies switch from nuclear to nucleolar specificity. To characterize the autoantibodies produced by RIIB−/−yaa mice and the nucleolar antigens that react with these antibodies, we have produced hybridomas by fusing RIIB−/−yaa mouse splenocytes with myeloma cells. Hybridoma H526 produced anti-nucleolar and hybridoma H333 produced anti-nuclear antibodies. H526 antibody binds RNA in an anti-RNA ELISA. Antibodies from H333 are IgG2a-κ whereas H526 are IgG2a-λ. For sequence analysis, variable regions of heavy chain (VH) and light chain (VL) of H526 and H333 antibodies were amplified by PCR from respective cDNAs, cloned, and sequenced. The VH and VL region sequences of both the antibodies showed high homology with anti-nuclear antibodies, chromosome 12 of C57BL/6 mouse genome, and the J558 immunoglobulin variable regions. There were only a few amino acid changes in the variable regions. The functional roles of the amino acid changes and the pathogenecity of the antibodies are under investigation. H526 nucleolar antibodies pull down five proteins from a nuclear extract as detected by Coomassie blue staining. Characterization of these nucleolar antigenic proteins is in progress.
Aged B6.SB-Yaa/J Mice develop an SLE-like Disease (130.11)
Authors: Bubier Jason A , Brown Aaron C. , Foreman Oded , Sproule Thomas J. , Roopenian Derry C.
Published: January 2023

Mouse models of SLE are considered pivotal in understanding the genetics of SLE along with identifying targets for therapy. Male mice of the BXSB/MpJ recombinant inbred strain, first described by Murphy and Roths in 1978, develop a spontaneous and severe SLE-like syndrome resulting in death averaging ~6 months of age, while females show only a very chronic syndrome. A mutant locus found on the Y chromosome is, therefore, appropriately named the Y-linked autoimmune accelerator (Yaa). Previous analysis have suggested that Yaa fails to induce an SLE-like disease in B6 mice, the caveat to this being that the SLE criteria analyzed were limited to serological analysis, histopathology, and mortality on mice up to 8–12 months of age. We report here that when mice of a significantly older age, 15–24 months, are compared to age-matched B6 controls, the B6.SB-Yaa/J mice clearly developed an SLE-like disease. This is characterized by significant splenomegaly, monocytosis, immune cells with an activation phenotype, the presence of homogeneous anti-nuclear antibody staining, glomerulonephritis, decreased weight and survival as well as a gene expression signature similar to BXSB/MpJ mice. These results suggest that Yaa alone is sufficient to induce an SLE-like syndrome in aged B6 mice.

Funded by NIH R21DK074463 and DK56597 to DCR, Arthritis Foundation to JAB.
CNS lupus in MRL/Faslpr mice (130.15)
Authors: Choi Jinyoung , Chahboune Halima , Hyder Fahmeed , Craft Joe
Published: January 2023

Neuropsychiatric involvement is common in patients with SLE; however, its etiology is incompletely understood. Fas-deficient MRL/Faslpr (MRL/Faslpr) mice develop a spontaneous autoimmune disease that is similar to human SLE, including cognitive and neurological deficits. We hypothesized that autoantibodies in MRL/Faslpr mice may promote central nervous system (CNS) injury. Antibody (Ig) deposition in the CNS and structural abnormalities were assessed by immunofluorescent microscopy and diffusion tensor imaging (DTI) using a 11.7T system. At 20 weeks of age, MRL/Faslpr mice exhibited dense deposits of IgG in the choroid plexus, whereas several other control mice, including Fas-intact MRL mice, Fas-deficient C57BL6/Faslpr mice, and membrane only IgM-transgenic MRL/Faslpr mice (B cell-intact, but incapable of Ig secretion) did not reveal IgG staining, suggesting spontaneous Ig entry into the CNS in this lupus-prone mouse. DTI analysis demonstrated that MRL/Faslpr mice exhibited significantly increase in dorsal-ventral directional anisotropy in the hippocampus, compared to other control mice, suggesting these mice may have a structural abnormality in the hippocampus. Future work will focus on the role of Toll-like receptors in the Ig deposition and subsequent DTI alteration at distal regions of hippocampus in the MRL/Faslpr mice.

Funding sources: Arthritis Investigator Award and Arthritis Innovative Grant, Arthritis Foundation
Do autoreactive memory B cells form as a consequence of extrafollicular activation? (130.18)
Authors: Sweet Rebecca , Herlands Robin , Shlomchik Mark
Published: January 2023

Memory B cells could be responsible for chronic autoimmunity, as suggested by results in SLE patients and SLE-prone mice: The frequency of AM14 RF transgenic B cells fluctuates in the peripheral blood of aged MRL/lpr mice. Activation of RF B cells in autoimmune MRL/lpr mice results in an extrafollicular response that undergoes somatic mutation. Lineage trees of RF clones suggest mutation upon secondary antigen exposure. We recently found that this response can be recapitulated in AM14 Tg mice that were exposed to IgG2a anti-chromatin Abs. The mechanism controlling extrafollicular activation and whether this response leads to a form of memory is unclear. When AM14 heavy chain knock-in B cells are transferred into non-transgenic recipients, followed by exposure to IgG2a anti-chromatin, an extrafollicular response that includes class switch ensues. We are tracking the development and function of these cells over time to determine if autoreactive resting memory B cells form, or alternatively, whether the maintenance of autoreactive B cell clones requires continuous autoantigen exposure and activation.

Supported by 1F31AI071694-01 (RAS) and AI36529 (MJS)
Identification of a single gene differentially expressed between males and females in both normal and lupus-prone mice (130.20)
Authors: Jorgensen Trine N. , Kappler John W. , Marrack Philippa
Published: January 2023

Systemic lupus erythematosus is an autoimmune disease with a pronounced female bias. In (NZB x NZW)F1 mice (BW), a model of spontaneous lupus-like disease, females also develop disease with a higher incidence than males, and it has been shown that sex hormones play a pivotal role. In humans, it is thought that disease development is a result of unknown environmental factors acting on a complex genetic background. We hypothesized that genes differentially expressed between males and females act in addition to lupus susceptibility genes to exert disease development in females. As autoantibody production by plasma cells is the main culprit of lupus-like disease in BW mice we focused our studies on B cells.

We analyzed the gene expression pattern of splenic B cells isolated from 9 wk old normal (B6 and Balb/c) and lupus-prone (BW) male and female mice. The results showed that 55 genes were differentially expressed between males and females in all three strains. To evaluate whether any of these genes were intrinsically different, we sorted splenic B cells from pre-pubertal (4 wk) male and female BW mice and compared genes differentially expressed in these mice with the previous data. Even less genes were found differentially expressed and only one gene, Sfi1, was consistently increased in females at both ages, suggesting that, in contrast to most of the other genes identified in 9 wk old mice, this gene is not regulated by sex hormones.
IL-17 promotes germinal center response and AID-regulated pathogenic autoantibody production in autoimmune BXD2 mice (130.21)
Authors: Hsu Hui-Chen , Yang PingAr , Wu Qi , Chen Jian , Wang John , Job Godwin , Stanus Andrea L , Lorenz Robinna G , Kolls Jay K , Mountz John D
Published: January 2023

We have found dramatically elevated levels of activation-induced cytidine deaminase (AID) in the B cells of generalized autoimmune BXD2 mice that develop spontaneous erosive arthritis and glomerulonepheritis. Pathogenic autoantibodies cloned from these mice exhibit extensive somatic hypermutation and the majority of them have undergone class-switch recombination, suggesting that AID activity plays an important role in the production of pathogenic autoantibodies. Sera from BXD2 mice exhibited significantly elevated IL-17, but not IFN-γ or IL-4, compared to B6 mice. FACS analysis indicates that CD4 T cells in BXD2 mice are preferentially polarized to Th17 cells and these cells also express the early activation marker, CD69, in vivo in BXD2 mice. The B cells from BXD2 mice express high levels of IL-17 receptor (IL-17R) and immunohistochemistry staining shows that B cells are in close contact with the IL-17+ cells in the spleen. Administration of AdIL-17 to 1-mo-old B6 and BXD2 mice induced the expression of AID, promoted development of GCs, and enhanced the sera titers of antibodies. In contrast, administration of AdIL-17R:Fc to 10-mo-old BXD2 mice suppressed the expression of AID, down-modulated the percentage of PNA+Fas+ GC B cells, and suppressed the sera titers of antibodies. Our results provided a unique pathogenic function of IL-17 in that it positions IL-17 as a critical cytokine that regulates GC B cell response and the production of pathogenic autoantibodies.
The NOD.H2h4 mouse shows characteristics of human Sjogren’s Syndrome (130.26)
Authors: Burek C. Lynne , Talor Monica V. , Sharma Rajni B. , Rose Noel R.
Published: January 2023

The objective of this study was to identify critical features of the NOD.H2h4 mouse as an appropriate model to study the pathogenesis of human Sjögren’s Syndrome. Salivary and lacrimal glands of male or female mice at different age groups were examined histologically for cellular infiltration. Sera were examined for ANA by indirect immunofluorescence on Hep2 slides and for SSA/Ro and SSB/La antibodies by ELISA. Salivary function was evaluated by measuring saliva volume following pilocarpine stimulation. At 20–24 weeks, female mice showed more frequent salivary gland infiltration than males ( 65% vs 40%), whereas male mice developed more frequent lacrimal gland infiltration (100% vs 38%). By one year, the frequency was similar in both sexes. ANA frequency increased with age, with approximately 10% of animals with ANA at six weeks, to about 80 % at 20 weeks. ANA pattern was primarily speckled. Antibodies to SSA/Ro and SSB/La were found at higher levels in year old NOD.H2h4 mice (Mean ELISA OD 0.4 ± 0.17 ) compared to 10 week old NOD.H2h4 (0.1 ± 0.14) or BALB/c mice (-0.14 ± 0.16), (p<0.001) with females greater than males. Antibodies to SSA/Ro and SSB/La increased with age from 10 weeks to 20 weeks. Salivary flow following pilocarpine stimulation was reduced in NOD.H2h4mice compared to control mice (146.7ul ± 56.4ul vs 198.8 ul ± 19.3ul , p = 0.03). The NOD.H2h4 mouse exhibits both salivary and lacrimal gland infiltration, high frequency of ANA with a high proportion of SSA/Ro and SSB/La autoantibodies, and decreased salivary secretion. Taken altogether, this mouse strain appears to be an ideal candidate to study the pathogenesis of Sjögren’s syndrome.
CD8 T cells Play a Critical Role in Primary Biliary Cirrhosis of dnTGFbetaRII Mice (130.28)
Authors: Yang Guo-Xiang , Lian Zhe-Xiong , Chuang Ya-Hui , Moritoki Yuki , Ansari Aftab A. , Flavell Richard A. , Ridgway William M , Gershwin M.Eric
Published: January 2023

Human primary biliary cirrhosis (PBC) is characterized serologically by antimitochondrial antibodies (AMA) and histologically by an intense T cell portal infiltrate that destroys bile ducts. We have taken advantage of the PBC-like disease exhibited by dnTGFbetaRII mice and focused our attention on the liver T cell infiltrate. One major advantage to this animal model is the ability to perform cell transfer. Therefore to address the issue of T cell effector mechanisms, we isolated dnTGFbetaRII splenic CD4+ or CD8+ T cells and transferred these populations into Rag1 k/o mice. Importantly, the CD8+ T cells transfer group demonstrated a significant expansion of T cells and the presence of portal tract infiltrates in recipient mice. In contrast, although CD4+ T cells did expand in the recipient group, they did not home or focus within the portal tracts. Our results demonstrate that the impaired TGFbeta signaling pathway in these mice leads to a CD8 cytotoxic T cell population that plays a critical role in biliary cell damage. These data have implications not only for understanding TGFbeta signaling and autoimmunity in these mice, but also in developing appropriate focused immunotherapy to prevent biliary damage.
Abrogation of lupus nephritis in activation-induced cytidine deaminase-deficient MRL/lpr mice (130.3)
Authors: Jiang Chuancang , Foley Julie , Clayton Natasha , Kissling Grace , Jokinen Micheal , Herbert Ronald , Diaz Marilyn
Published: January 2023

Autoantibodies in systemic lupus erythematosus (SLE) patients and in mouse models of SLE are frequently found to be class switched and somatically mutated. To examine directly the role of hypermutated and isotype-switched antibodies in the pathogenesis of SLE, we generated a new MRL/lpr mouse strain deficient in the activation-induced cytidine deaminase (AID). Unlike their AID wild-type littermates, AID-deficient MRL/lpr mice not only lacked autoreactive IgG antibodies, but also experienced a dramatic increase in the levels of autoreactive IgM. This phenotype translated into a near abrogation of glomerulonephritis, minimal mononuclear cell infiltration in the kidneys, and a dramatic increase in survival, indicating that isotype-switched high affinity autoantibodies are required for the development of lupus nephritis. The dramatic decrease in mortality to levels well below levels previously reported for MRL/lpr mice lacking secreted antibodies, suggests that autoreactive IgM antibodies not only are insufficient in promoting kidney disease in MRL/lpr mice, but may also play a protective role.

This research was supported by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences.
The silent chemokine receptor D6 is required for generating T cell responses that mediate experimental autoimmune encephalomyelitis (130.33)
Authors: Liu Liping , Graham Gerard J , Hu Taofang , Lira Sergio A , Sasse Margaret , Canasto-Chibuque Claudia , Cook Donald N
Published: January 2023

D6, a promiscuous non-signaling chemokine binding molecule expressed on lymphatic endothelium, internalizes and degrades CC chemokines, and D6−/− mice demonstrated increased cutaneous inflammation following topical phorbol ester or complete Freund’s adjuvant (CFA) injection. We report that D6−/− mice were unexpectedly resistant to induction of experimental autoimmune encephalomyelitis (EAE), due to impaired encephalitogenic responses. Following induction with myelin oligodendroglial glycoprotein (MOG) peptide 35–55 in CFA, D6−/− mice showed reduced spinal cord inflammation and demyelination, with lower incidence and severity of EAE attacks, compared to D6+/+ littermates. In adoptive transfer studies, MOG-primed D6+/- T cells equally mediated disease in D6+/+ or D6−/− mice, while cells from D6−/− mice transferred disease poorly to D6+/- recipients. Lymph node cells from MOG-primed D6−/− mice showed weak proliferative responses and made reduced IFN-gamma, but normal IL-5. CD11c+ dendritic cells accumulated abnormally in cutaneous immunization sites of D6−/− mice. Surprisingly, D6, a ‘silent’ chemokine receptor, supports immune response generation.
Streptozocin-induced diabetes accelerated by B cells transgenic for immunoglobulin heavy chain recognizing N-acetyl-D-glucosamine (130.35)
Authors: Dizon Brian , Greenspan Neil S. , Kearney John F
Published: January 2023

To determine mechanisms of autoreactive B cell selection and their involvement in the pathogenesis of autoimmune diseases, we generated a novel immunoglobulin (Ig) transgenic (Tg) mouse line, HGAC 39. In these mice, B cells express an Ig heavy chain with specificity for N-acetyl-D-glucosamine (GlcNAc) found on the cell wall of Group A Streptococcus (GAS). Tg B cells responded to iv immunization by migrating from the marginal zone to the TB border, and expanding and differentiating to plasmablasts at extrafollicular foci. Since GlcNAcylated proteins are abundant within eukaryotic cells, we hypothesized that B cells with anti-GlcNAc specificities contribute to autoimmune disease by presenting GlcNAc-modified proteins to diabetogenic CD4+ and CD8+ T cells. We induced autoimmune diabetes in Tg C57BL/6 male mice with multiple low doses of streptozocin (STZ), a β cell-specific toxin that increases GlcNAcylation of intracellular proteins, and demonstrated that the presence of HGAC 39 B cells accelerates the onset of hyperglycemia and pancreatic infiltration by HGAC 39 B cells, CD4+, and CD8+ T cells. These changes were not accompanied by increases in anti-GlcNAc plasma cells or serum antibodies. Taken together, our data suggest that HGAC 39 B cells that respond to GAS immunization can also contribute to STZ-induced diabetes by mechanisms other than autoantibody secretion.
Countervailing actions of myeloid and plasmacytoid dendritic cells control autoimmune diabetes (130.36)
Authors: Saxena Vijay , Ondr Jennifer K , Magnusen Albert F , Munn David , Katz Jonathan D
Published: January 2023

Islets specific CD4+ T Cells receive instructional signals from antigen bearing MHC class II-expressing antigen presenting cells (APC), yet the influence of APC on autoimmune diabetes remains elusive. Herein we delineate the direct in vivo necessity for macrophages and dendritic cells in Type 1 Diabetes of the non-obese diabetic (NOD) mouse by using diphtheria toxin-mediated conditional abaltion. We find that specific loss of macrophages had no impact on diabetes or insulitis in both spontaneous and transfer model. Loss of both myeloid (mDC) and lymphoid DC (ly DC) led to protection of NOD mice from diabetes and accompanying loss of T cell activation, while specific loss of plasmacytoid DC (pDC) and Indoleamine 2,3,dioxygenase (IDO) led to breakdown in regulatory control and accelerated insultis. These data imply that autoimmune diabetes is regulated by imbalance of mDC and pDC function in vivo.
Elucidating The Role Of Insulin Receptor In Type-I Diabetes By Developing An IR Transgenic Mouse Model (130.37)
Authors: Gupta Nirdesh K , Bretz James D , McInerney Marcia F
Published: January 2023

In the nonobese diabetic (NOD) mouse, type1 diabetes (T1D) is an autoimmune disease characterized by insulitis and T cell-mediated destruction of pancreatic islet β cells. The importance of insulin receptor (IR) expression in the pathogenesis of diabetes was examined since it was shown that IR is a chemotactic receptor capable of directing cell movement in response to insulin. Our published data shows that IR High/+ T cells aggressively transfer insulitis and diabetes while IR Low/− T cells are capable of neither. Therefore, IR may function to deliver T cells to the islet. Recently, it has been shown in NOD mice that benign insulitis can occur in the absence of T cell-MHC/Ag interactions. The object of this work is to test whether T cells overexpressing IR, can move into the pancreas based on insulin chemotaxis alone, without being targeted to the pancreas by antigenic specificity. To that end a transgenic mouse will be made wherein FLAG tagged IR expression will be targeted to all T cells. If IR expression is a mechanism to move T cells into the islet then down regulation of IR expression or blocking of chemotactic IR activity specifically in T cells, will provide a new therapeutic target to block cell movement into the pancreas, thus preventing diabetes.
CD4+ T cells mediate autoimmune disease in a new transgenic murine model induced by U1 RNA and U1-70kD ribonucleoprotein self antigens (130.4)
Authors: Hoffman Robert W , Greidinger Eric L , Zang Yun , Jaimes Kimberly , Martinez Laisel , Fernandez Irina
Published: January 2023

C57BL/6 mice transgenic (Tg) for the human major histocompatibility complex gene HLA-DR4 were immunized with U1 RNA and a fusion protein spanning the RNA binding domain of the U1-70kD polypeptide, control fusion protein, with or without U1-RNA, or saline. Complete serologic and histologic analyses were performed on all mice. CD4+ T cells and APCs were isolated from immunized mice using immunoaffinity columns and tested for proliferation. Either freshly isolated CD4+ T cells from immunized mice or T cell lines were adoptively transferred to unmanipulated HLA-DR4 Tg or RAG−/− mice. Freshly isolated CD4+ T cell from immunized DR4 Tg mice proliferated in response to U1-70kD and to synthetic peptides from the RNA binding domain region of the protein in the presence of irradiated autologous APCs. We found that CD4+ T cells specific for U1-70kD were able to transfer disease to unmanipulated syngeneic mice. Furthermore, using purified CD4+ T cells or T cell lines specific for U1-70kD we were able to transfer disease to RAG−/− mice in the absence of detectable B cells or autoantibodies.

In conclusion, we have developed a new model of systemic autoimmunity that is induced following a single exposure to U1-RNA and its associated RNA binding protein. Anti-U1-70kD autoantigen-specfic CD4+ T cells appear to medicate pathogenesis of disease in this model and can do so in the absence of detectable B cells or autoantibodies.

This work was supported by NIH award AR43308 (RWH), the Lupus Foundation of American (RWH) and Department of Veterans Affairs Merit Review awards (RWH & ELG).
Polarized TCR-Transgenic Th17 Cells Resemble Th1 Cells in Their Capacity to Adoptively Transfer Ocular Inflammation, but Differ in Other Biological Activities (130.44)
Authors: Cox Catherine A , Shi G. , Yin H. , Vistica B.P. , Wawrousek E.F. , Chan C-C. , Gery I.
Published: January 2023

The role of Th17 lymphocytes in immunopathogenic processes has been well established, although little is known about their mode of action and other biological activities. Here, we prepared polarized populations of TCR-transgenic (Tg) Th1 or Th17 cells specific against hen egg lysozyme (HEL) and compared each line for its capacity to adoptively transfer ocular inflammation into Tg recipients expressing lens-associated HEL, as well as for other biological activities. To establish each line, naive CD4+ cells were cultured in the presence of HEL and "cocktails" containing cytokines and antibodies known to drive polarization to Th1 or Th17. Successful polarization of Th1 and Th17 lines was confirmed by intracellular expression and release into the culture medium of IFNγ or IL17, respectively. Surface marker expression was determined by flow cytometry of cultured cells. Recipient eyes were examined histologically. Polarized cells were also tested for their capacity to induce splenomegaly in recipient mice (Chen et al JI 2006). Our results showed that Th1 and Th17 cells were similarly immunopathogenic, transferring ocular inflammation at doses as low as 0.2 million. Histological comparison of eyes from each group, however, revealed slight although discernible differences between Th1 and Th17 recipients. The two populations also differed in their expression of surface markers, particularly in CD49d and CD62L. Finally, Th17 cells were remarkably less efficient than Th1 cells in their capacity to induce splenomegaly in recipient mice.
Potential regulatory role for CNS-specific, autoreactive CD8+ T-cells in autoimmune demyelination (130.7)
Authors: York Nathan Ronald , Mendoza Jason P , Benagh Andrew , Karandikar Nitin J
Published: January 2023

A great deal of our current knowledge about the immune pathogenesis of multiple sclerosis (MS) derives from work in its murine model, experimental autoimmune encephalomyelitis (EAE). The vast majority of studies in MS and EAE have focused on the role of CD4+ T cells, with the underlying assumption that MS, like EAE, is a CD4+ Th1-mediated and Th2-regulated autoimmune disease. However, several reports from us and others have implicated roles for both CD4+ and CD8+ T-cells in these diseases. In this study, we specifically dissected the role of autoreactive, CNS-specific CD8+ T-cells in EAE. Similar to our observations in MS, we saw robust induction of CNS-specific CD8+ T-cell responses in several established models of EAE (B6/MOG35-55; SJL/PLP139-151; B10.PL/MBPAc1-11). Following in vitro activation in bulk splenocyte cultures, MOG35-55-specific CD4+ T-cells, but not CD8+ T-cells, were capable of adoptively transferring EAE to naïve B6 mice. Interestingly, transfer of MOG-stimulated, but not OVA-stimulated, CD8+ T-cells resulted in inhibition of EAE induction, suggesting a regulatory, rather than pathogenic, role for these cells in this model. In conclusion, our studies reveal a novel regulatory function for CNS-specific CD8+ T-cells that are reproducibly generated during the induction of EAE.

This research was supported, in part, by grants from the National Institutes of Health and the National MS Society.
Spontaneous Gluten Dependent Blistering in DQ2 transgenic mice (130.9)
Authors: Marietta Eric Vincent , Rashtak Shadi , Pittelkow Mark R , David Chella S , Murray Joseph A
Published: January 2023

Background/Aim: Dermatitis Herpetiformis is the autoimmune skin manifestation of celiac disease, and similar to celiac disease, is tightly associated with HLA-DQ2 and HLA-DQ8. Previously we have generated a mouse model of DH using DQ8 transgenic mice on an NOD background. Herein, we describe a similar DH-like blistering disease that spontaneously develops in DQ2 transgenic mice.

Methods: Transgenic mice that express DQ2 on a background deficient in endogenous MHC II (Aeo) were raised on a gluten containing (GC) chow or a gluten free chow .

Results: When raised and maintained on a gluten containing chow, approximately 15% of these Aeo DQ2 mice spontaneously developed blisters on the ears and tails at about six months of age. Control Aeo mice that were also raised on a gluten containing chow did not develop this blistering disease during their lifetime. Hematoxilyn and eosin staining revealed subepidermal blisters. Immunohistochemistry revealed IgA deposits at the dermal/epidermal junction in perilesional ear tissue. However, when these mice were raised and maintained on a gluten free diet, these mice did not develop blisters.

Conclusions: This blistering disease is a gluten dependent disease that has many of the elements typically associated with Dermatitis Herpetiformis, most especially the association with the DQ2 gene.
Vaccination with Collagen-Pulsed Immature Dendritic cells Significantly Delayed the Onset and Lessened Severity of Spontaneous Polychondritis in HLA-DQ Transgenic Mice (131.10)
Authors: Sidhu Maninder , Bradley David S
Published: January 2023

Immature dendritic cells (iDCs) have a tolerogenic potential due to low expression of costimulatory cell surface molecules required for antigen presentation and induction of an effective immune response. Here we report that injection of iDCs pulsed with chick type II collagen (CII) significantly delayed the onset and suppressed the severity of spontaneous polychondritis in HLA-DQ6αβ8αβ tg mice. Bone marrow derived iDCs were purified, pulsed in vitro with CII and transferred to 6 wk old HLA-DQ6αβ8αβ tg mice. Controls received either un-pulsed iDCs or PBS. Both control groups developed polyarthritis at 3.5 months as we have previously reported. However, none of the mice receiving CII-loaded iDCs displayed any clinical signs of disease until 5.5 months of age, indicating the ability of the DC vaccine to significantly delay the onset of the disease. The severity of disease in CII-pulsed iDC immunized test mice remained less than controls. Interestingly, after 6.5 months of age the incidence of polyarthritis in the CII-loaded iDC group gradually increased from 25% to 87%. Total IgG levels were elevated in the CII-loaded iDC test group. None of the mice developed significant CII-specific IgG responses, indicating that the CII-loaded iDCs were not inducing a CII specific immune response. This is the first evidence of iDC therapy controlling polychondritis and suggests that iDC vaccination may provide a novel tool in the treatment of human inflammatory autoimmune disease such as relapsing polychondritis and rheumatoid arthritis.
Suppression of Collagen-Induced Arthritis with Type-II collagen peptide linked Soluble MHC II (I-Aq) molecules (131.12)
Authors: Park Yoon-Kyung , Jung Sundo , Park Se-Ho
Published: January 2023

In collagen-induced arthritis (CIA) of mouse, self-reactive T cells recognize a peptide antigen from type II collagen (CII). CII is of particular interest, as an autoimmune response to this protein leads to CIA in mice, rats, and primates. Activation of T cells is believed to be an important pathogenic factor in autoimmune disease. So, T cells have become a focal point of study for the development of novel therapeutic approaches to the treatment of autoimmune disease. In this study, we evaluated the efficacy and mechanism of recombinant MHC II molecules in regulation of the antigen-specific T cell clones by using mouse I-Aq, combined with an auto-antigen peptide from type II collagen (CII260-274) in CIA model. It was found that recombinant I-Aq/CII260-274 molecules not only activate CII-specific T cell clone but also inhibit the same clone in vitro according to the condition of stimulation. Furthermore, development of CIA in mice was successfully prevented by the in vivo injection of a recombinant soluble I-Aq/CII260-274. Thus, recombinant soluble MHC II molecules, complex with the immuno-dominant self peptide, offer a new possibility for the treatment of chronically active autoimmune inflammation such as rheumatoid arthritis.
Determination of genetic factors influencing susceptibility to disease and response to interferon-B in multiple sclerosis (131.16)
Authors: O’Doherty Catherine , Hawkins Stanley , Hutchinson Michael , Vandenbroeck Koen
Published: January 2023

Treatment strategies of multiple sclerosis (MS) aim to slow progression of the disease and provide symptomatic relief. Interferon B (IFN B) is the most prominent disease modifying drug used in the treatment of MS. It has been shown to decrease relapse rates and delay progression of the disease. There are currently three proprietary IFN B products on the market; Avonex and Rebif (IFN B1a), and Betaferon (IFN B1b). Despite demonstrated efficacy, these products are not without their problems. There is considerable variation in response, with an estimated 30–50% of patients showing treatment failure, and high treatments costs. In essence, treatment is on a trial and error basis. To date there is no method of treatment outcome prediction at onset of treatment. In this study we have screened approximately 60 polymorphisms in over 30 candidate response genes, based on notions of their known function and the literature. We selected SNPs in the promoter region, hypothesising that polymorphisms may modify IFN inducibility. Additionally non-synonymous SNPs and those potentially affecting splicing, were also selected. Polymorphisms were typed in DNA samples from 260 Irish patients classified as responders or non-responders. A cohort of 120 healthy controls was also included in the study to determine whether these SNPs influenced susceptibility to MS.