bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential production of reactive oxygen intermediates by TCR stimulation in SLE T cells (129.10)
Authors: Khan Islam U. , Michalek Ryan D. , Giri Banabihari , Grayson Jason M. 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a protean autoimmune disease characterized by a wide array of intrinsic T cell defects. Reactive oxygen intermediates (ROI) modulate T cell activation, cytokine production, and proliferation at multiple levels. The purpose of this study was to measure T cell proliferation and ROI production in SLE versus healthy control T cells. CD4+ and CD8+ T cells were isolated by negative selection magnetic micro-beads. We characterized the memory phenotype and activation markers on the cells from both groups by flow cytometry. CD62Lhigh and CCR7high were the most abundant subsets in CD8+ T cells from healthy subjects. The frequency of CCR7highCD8+ T cells in SLE patients was significantly lowered compared to healthy subjects (p=0.0001). For proliferation and ROI studies, cells were stimulated with anti-CD3 and anti-CD28 antibodies. CFSE staining following activation revealed that SLE CD8+ T cells had decreased division in response to TCR-mediated stimulation as compared to healthy subjects. Both subsets of SLE T cells showed increased ROI in response to activation. Interestingly, among the SLE subjects, the maximum increase in H2O2 production was recorded in the CD8+ T cells of one patient with severe SLE disease. In conclusion, we demonstrate that T cells from SLE patients produce increased levels of ROI upon stimulation implicating ROI in SLE pathogenesis.
Supported by NIH RO1-AR39501
The Yaa locus Confers Apoptosis, NF-κB and Rapamycin Signaling Resistance Through Multiple Receptors in C57BL/6J Mice (129.12)
Authors: Brown Aaron C , Bubier Jason A , Sproule Thomas J , Christianson Greg J , Roopenian Derry C 
Published: January 2023

Abstract
BXSB/MpJ mice develop a spontaneous SLE-like disease that is remarkably accelerated in males, due to the presence of the Y-linked-autoimmune accelerator locus (Yaa). Recent insights into the molecular nature of the Yaa mutation were provided when an additional copy of 17 distally located X chromosome genes were discovered on the Y-chromosome of Yaa mice. Included within this region is the gene TLR7, a toll receptor which binds ssRNA and is functionally expressed on B-cells and monocyte derived lineages. Previous reports have suggested that the SLE-like disease phenotype conveyed by Yaa is due to the duplication of TLR7, which causes a shift toward anti-nucleolar antigens in B6.FcγRIIB(−/−). We show here that B6, BXSB and MRL mice that carry Yaa rarely display the anti-nucleolar antibody pattern. We found that B cells of Yaa mice display novel phenotypes consistent with increased rapamycin and NF-κB signaling through receptors other than TLR7, including TLR4, TLR9, and the BCR. In addition, Yaa B cells are resistant to anti-IgM induced apoptosis. These results demonstrate
that the copy number of a single receptor can influence the strength of signaling through other receptors and/orthat additional genes within the duplicated region contribute toward the Yaa phenotype.
Funded by NIH R21DK074463 and DK56597 to DCR, Arthritis Foundation to JAB.
LOSS OF RIP2 EXACERBATES MURINE EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (129.16)
Authors: Fairhead Todd , Fenutria Rafael , McCully Michelle L. , Lemke Caitlin D. , Davis Brianne , Lozano Francisco , Strejan Gill , Madrenas Joaquin 
Published: January 2023

Abstract
Receptor interacting protein 2 (RIP2), a putative serine-threonine kinase, is a modulator of the activation of both innate and adaptive immune responses. RIP2-deficient mice have impaired NF-κB signaling and defects in Th1-mediated immune responses. Since activated NF-κB signaling in hematopoietic tissue and in the central nervous system (CNS) has been implicated in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), we hypothesized that deficient expression of RIP2, a regulator of the NF-κB pathway, would affect the course of EAE.
To test this hypothesis, we induced EAE with MOG35-55 peptide, in C57Bl/6 wild type (WT) and RIP2−/− mice. We found that RIP2−/− mice exhibited exacerbated EAE as compared to WT mice, with both a higher clinical score and increased demyelination. Disease induction in both genotypes was T cell-dependent as demonstrated by adoptive T cell transfer. However, EAE in RIP2−/− mice did not have an accelerated course and did not show increased T cell priming ex vivo. Rather, the exacerbated EAE in RIP2−/− mice was associated with a higher percentage of IL-17-producing T cells, both in the periphery and in the CNS, while the percentage of IFN-γ-producing cells was the same in WT and RIP2−/− mice.
Our data demonstrate that RIP2 is a key in vivo regulator of Th17-associated autoimmunity in the CNS. Such a role is likely due to its activity on the NF-κB pathway.
Reduced expression of NKp46 on CD56dim natural killer cells from MS patients (129.21)
Authors: Edwan Jehad H , Bielekova Bibiana 
Published: January 2023

Abstract
Natural killer (NK) cells possess innate immune responses and play a crucial role in regulating autoimmune T cells. NKp46 is a natural cytotoxicity activating receptor exclusively expressed on NK cells. NKp46 plays a pivotal role in NK cell functions because it mediates the lysis of many human tumor cells lines, as well as infected cells. We investigated the expression of NKp46, CD62L and CD44 on NK cells from Multiple Sclerosis (MS) patients and healthy donors (HD). Apheresis blood samples were obtained from MS patients and HD, PBMCs were separated by density gradient centrifugation with lymphocytes separation medium. NK cells were negatively selected using MACS NK isolation kit, and cultured with or without 1 μg/ml influenza vaccine (Flu HA) for 4 hours. Using 6 color flow cytometry, we examined the CD56+ CD3− cells for the expression of NKp46, CD62L and CD44. We observed that MS patients’ NK cells express unusually low frequency of CD56dim CD44+ NKp46+ cells compared to HD. The levels of CD56bright cells expressing CD44+ CD62L+ NKp46+ were comparable among MS patients and HD. Four hours after incubation with Flu HA, the two CD56dim and CD56bright populations expressing NKp46+ completely disappeared leaving one population expressing only CD56dim CD44+. Our findings suggest that CD56dim CD44+ NKp46+ might be involved in the protection against development of MS.
IFNγ-dependent mechanisms of activation-induced cell death control the progression of autoimmune myocarditis (129.32)
Authors: Barin Jobert G. , Cihakova Daniela , Kimura Miho , Talor Monica V. , Rose Noel R. 
Published: January 2023

Abstract
Interferon gamma (IFNγ) exerts a suppressive effect on experimental autoimmune myocarditis (EAM) in mice, as well as other models of organ-specific autoimmune disease. We have sought to determine the mechanisms by which IFNγ mediates this effect, which may involve the apoptosis of autoreactive lymphocytes by activation-induced cell death. IFNγ knockout and wild type control BALB/c mice were immunized with the cardiac myosin-derived peptide MyHCα614-629 to induce EAM. At day 21 post-immunization, we observed a marked decrease in the activity of Caspase 8 in splenic CD4+ T cells in IFNγ−/− mice with EAM. We further found that this decrease in CD4+ cell Caspase 8 activity could be recapitulated in an antigen-specific manner in vitro by stimulation of spleen cells from IFNγ−/− mice with the MyHCα614-629 peptide. These differences in Caspase 8 activity are paralleled by decreases in the ability of MyHCα614-629-reactive CD4+ cells from IFNγ−/− mice with EAM to apoptose after in vitro stimulation with the peptide. These data support the hypothesis that IFNγ acts through regulating apoptotic mechanisms in autoreactive lymphocytes to limit the progression of autoimmune disease.
(Supported by 2R01 HL67290)
CD24 inhibits thymic deletion of myelin antigen-specific T cells (129.34)
Authors: Carl Joseph W , Joshi Pramod , Whitacre Caroline C , Liu Yang , Bai Xue-Feng 
Published: January 2023

Abstract
CD24 is a glycosylphosphatidylinositol (GPI) anchored cell surface glycoprotein that is expressed in hematopoietic cells and cells of the central nervous system (CNS). We have previously shown that CD24 is required for the induction of experimental autoimmune encephalomyelitis (EAE), an experimental model of human disease multiple sclerosis. In CD24−/− mice, normal levels of myelin oligodendrocyte glycoprotein (MOG) specific T cell were primed, however these T cells were non-pathogenic. To understand this issue, we bred CD24−/− mice with 2D2 TCR transgenic mice, which bear TCR specific to MOG, and generated 2D2 TCR transgenic mice with or without CD24. Here we show that 2D2 TCR transgenic mice with CD24-deficiency (2D2/CD24−/−) have remarkably withered thymus. In peripheral lymphoid organs, transgenic T cells from 2D2/CD24−/− mice have an immature phenotype (CD4−CD8−), do not respond to MOG peptide stimulation, and fail to cause autoimmune inflammation in the CNS and optical nerves. In contrast, OT-2 TCR transgenic mice with CD24 deficiency (OT-2/CD24−/−), which bear TCR specific to chicken ovalbumin (OVA), have normal thymus and their peripheral T cells have a normal response to OVA peptide. These data suggest that CD24 inhibits thymic deletion of myelin antigen, but not foreign antigen-reactive T cells.
The LFA1-ICAM2-ezrin-Akt pathway in mediating fibroblast-like synoviocyte proliferation in rheumatoid arthritis (129.40)
Authors: SINGH KARNAIL , COLMEGNA INES , HE XIAOWEN , WEYAND CORNELIA M , GORONZY JORG J 
Published: January 2023

Abstract
In rheumatoid arthritis (RA), the synovium is infiltrated by mononuclear cells that influence the proliferation and activation of fibroblast-like synoviocytes (FLS) through soluble mediators as well as cell-cell contact. To identify receptor-ligand pairs involved in this cross-talk, we cultured T cells with FLS lines isolated from RA synovium. T cells induced increased phospho-Akt (Ser473) in FLS, leading to downstream phosphorylation of GSK-3, FoxO1/3a and MDM-2 and enhanced in vitro FLS proliferation. Akt phosphorylation and enhanced FLS proliferation did not require MHC/antigen recognition, but were inhibited by antibodies against ICAM-2, 3 and CD11a. Antibodies to ICAM-1 were not effective. The selective role of the LFA-1 - ICAM-2/3 pathway was confirmed by the finding of increased ezrin phosphorylation at Tyr353 which is known to be downstream of ICAM2/3 and supports cell survival through Akt activation. CD28- T cells, which were present in significantly higher frequencies in RA patients than in healthy controls, had higher CD11a cell surface expression than naïve or regular memory T cells and were more potent to induce Akt phosphorylation in FLS. These findings identify ICAM-2/3 as therapeutic targets in RA allowing for a more selective intervention than the LFA-1 inhibition explored in clinical studies.
(Supported by NIH grants AR41974, AR42527 and AI44142).
Immunohistochemical detection of autoantibodies in the plasma of patients with Autism to neural cells of the non-human primate (129.42)
Authors: Wills Sharifia , Cabanlit Maricel , Bennett Jeff , Ashwood Paul , Amaral David , Van de Water Judy 
Published: January 2023

Abstract
Autoantibodies directed to various ‘self’ neural proteins have been reported in a subgroup of subjects with autism spectrum disorders (ASD) and are proposed to contribute to abnormal neurodevelopment. Our group sought to identify the presence of autoantibodies to brain protein(s) in the plasma of subjects with ASD enrolled in a population based case controlled study. Groups examined include patients with ASD, typically developing age-matched controls, siblings of children with ASD, and children with other developmental disabilities. Immunohistochemical staining of coronal brain sections obtained from the cynomolgus monkey (Macaca fascicularis) was performed to identify specific cellular targets against which the ‘self’ reactive antibodies are directed. Immunohistochemical analysis has thus far been confined to the cerebellum. Strong immunoreactivity was noted in the plasma from 20% of children with autism towards what appear morphologically to be golgi cells in the granular layer of the cerebellum. Such staining was absent in plasma from the typically developing controls, but present in two siblings and one disease control. Identification of the brain-specific autoantigens will help illuminate which brain structures and/or neurodevelopmental processes may be affected in patients with ASD.
[These studies were funded by NIEHS grant 1 P01 ES11269-01 and the M.I.N.D. Institute.]
Activated Insulin-Reactive CD8 T cells in NOD mice Cause Diabetes. (129.46)
Authors: Wong F. Susan , Siew Lai Khai , Thomas Ian J. , Chapman Stephen , Viret Christophe , Wen Li 
Published: January 2023

Abstract
We have previously generated highly diabetogenic insulin-reactive CD8 T cell clones designated G9C8 from the NOD mouse. The aim of our study was to investigate T cell selection and activation of CD8 T cells from a TCR transgenic mouse expressing the cloned TCRαβ receptor of the G9C8 clone. The mice were crossed to TCR.Cα−/− mice so that all the T cells express the clonotypic TCR. There was a predominance of CD8+ single positive thymocytes and peripheral CD8 TCR transgenic cells, compared with CD4 T cells. Peripheral lymph node T cells express a naïve phenotype (CD44lo, CD62Lhi). They proliferate to the insulin B15-23 peptide, produce interferon-γ and are cytotoxic to targets coated with the insulin peptide. In vivo, the TCR transgenic mice developed insulitis but surprisingly spontaneous diabetes did not occur. However, the TCR transgenic mice can be induced to develop diabetes on immunization. They will also cause diabetes on adoptive transfer after activation. In summary, these mice provide a good model for studying selection and activation of insulin-reactive CD8 T cells in the NOD mouse.
Bim−/−Faslpr/lpr Mice Exhibit Accelerated Lymphadenopathy (130.13)
Authors: Weant Ashley E , Khan Islam U , Holbrook Beth C , Willingham Mark C , Grayson Jason M 
Published: January 2023

Abstract
Apoptosis plays a critical role in the development and homeostasis of the immune system. To determine whether loss of genes in both extrinsic and intrinsic cell death pathways would exacerbate defects in immune homeostasis we created Bim−/−Faslpr/lpr mice. Mice were born at the expected Mendelian ratios and appeared to develop normally. However, by 14 weeks of age they developed a massive lymphadenopathy compared to either single mutant or wildtype animals. Their kidneys contained massive lymphocyte infiltration compared to the other genotypes. We determined the apoptotic sensitivity of T cells from these animals and found that naïve T cells cultured in vitro were resistant to growth factor deprivation similar to the Bim mutants and activated T cells were resistant to activation induced cell death similar to the Faslpr/lpr mice. We next determined the ability of these cells to divide following TCR stimulation. Upon stimulation, CD8+ T cells from these animals had slightly decreased proliferation than cells from the other genotypes. However, the percentage of cells surviving through division is increased in the double mutant animals. In conclusion, although T cells from Bim−/−Faslpr/lpr mice exhibit similar in vitro survival as either parental strain, in vivo the number of lymphocytes is synergistically increased resulting in massive lymphadenopathy.
Supported by ACS- #RSG-04-066-01-MBC to Jason Grayson.
Activation of hepatic stellate cells during fibrosis: Comparison of the CYP2D6 model for autoimmune hepatits and CCl4 injection (130.2)
Authors: Christen Urs , Hintermann Edith , Christen Selina , Bayer Monika 
Published: January 2023

Abstract
Only little is known about the mechanisms of periportal fibrosis in the pathogenesis of human autoimmune hepatitis. We used the virus induced CYP2D6 model system to investigate the activation of hepatic stellate cells (HSC) and the kinetics of fibrosis in comparison with the CCl4-induced fibrosis model. CYP2D6 transgenic mice express the human Cytochrome P450 in the liver and develop liver damage upon Adenovirus-CYP2D6 infection. In the CYP2D6 model we found mostly subcapsular fibrosis resulting in the fusion of individual lobules (Sirius Red, Collagen I). At 10–12 weeks post-infection, weak periportal fibrosis became apparent. In contrast, in CCl4-treated mice the kinetic of extracellular matrix deposition was accelerated resulting in periportal fibrosis after 3–4 week of CCl4 administration. At later times, fibrosis was much more pronounced in CCl4-treated mice compared to virus-infected CYP2D6 mice but subcapsular fibrosis was not as dominat. Activation of HSCs could be detected by staining for α-smooth muscle actin (αSMA) in liver sections of both CCl4-treated mice and virus-infected CYP2D6 mice. In addition, isolation of HSCs revealed an enhanced activation status (decreased amount of oil droplets, de novo αSMA expression) in CCl4-treated mice and virus-infected CYP2D6 mice. Our data indicate that virus-infected CYP2D6 mice display subcapsular and periportal fibrosis that correlates with an activation of HSCs similar to CCl4-induced fibrosis. Thus, the CYP2D6 mouse is a good model system to further investigate the molecular mechanisms involved in fibrotic events during autoimmune hepatitis.
Chronic low-grade exposure to bacterial superantigens elicits sustained T cell expansion and multiorgan inflammation. Implications for autoimmunity. (130.22)
Authors: Rajagopalan Govindarajan , Smart Michele K , Grande Joseph P , David Chella S 
Published: January 2023

Abstract
Bacterial superantigens (BSAg) are produced by Staphylococcus aureus and Streptococcus pyogenes. BSAg bind directly to the MHC class II molecules without undergoing any processing and subsequently activate T cells in an antigen non-specific manner by interacting with the TCR Vβ region. BSAg have been implicated primarily in acute diseases such as food poisoning, toxic shock syndrome, etc. Since carrier strains of S. aureus and S. pyogenes can elaborate superantigens, there is a distinct possibility of chronic exposure to low doses of BSAg and it is imperative to study its outcome. As BSAg bind more avidly to human (HLA) than to murine (H-2) MHC class II, we delivered low concentrations (total of 10 μg/mouse) of BSAg to HLA-DR3 [staphylococcal enterotoxin (SE) B] or HLA-DQ8 [streptococcal pyrogenic exotoxin (SPE) A] transgenic mice through subcutaneously implanted miniosmotic pumps for 7 days.
Continuous exposure to low concentrations of BSAg resulted in a significant protracted expansion of CD4+ T cells expressing TCR Vβ8 (for SEB); Vβ8 and Vβ14 (for SPEA) with a concomitant decrease in percentage of CD8+ T cells expressing these TCR. T cells bearing TCR Vβ6 showed an inverse correlation (slight decrease in CD4+ and increase in CD8+ compartments with SEB and SPEA). Histological examinations of major organs showed intense perivascular infiltration in lungs, kidneys and liver along with inflammatory infiltrates in the cardiac valvular endothelium in mice bearing pumps delivering BSAg but not PBS. In conclusion, we have shown the immunological outcome of chronic exposure to low levels of BSAg using the HLA class II transgenic mouse model.
Generation of T-cell Hybridomas Specific for Bile Duct and Rhesus Rotavirus Antigens in a Murine Model of Biliary Atresia (130.27)
Authors: Tucker Rebecca , Gill Ronald , Mack Cara 
Published: January 2023

Abstract
Rhesus rotavirus (RRV)-inoculated neonatal BALB/c mice develop an immune mediated inflamation of both extra- and intraheptatic bile ducts which progresses to biliary obstruction and death by 3 weeks of age. The livers of diseased animals demonstrate increased numbers of T-lymphocytes along with elevated expression of Th1 type cytokines at both one and two weeks. Previous work in our laboratory has demonstrated that the liver infiltrating T-lymphocytes are both RRV-reactive and autoreactive to bile duct antigens. In addition, transfer of T-cells from diseased animals leads to inflamation of bile ducts in SCID mice. This inflamation occurs in the absence of virus, suggesting that bile duct antigens alone are capable of soliciting this response. In order to identify the antigens being targeted, and to determine if the autoreactivity arises via an epitope spreading or molecular mimicry mechanism, we have generated T-cell hybridomas from the livers of 2 week old diseased mice. Initial analysis of these hybridomas reveals four classes of hybridomas.
TCR positive but non-reactive to tested antigens,RRV-specific,bile duct-specific, anddual reactive to both RRV and bile duct antigens.
Further study of these hybridomas should clarify what mechanism is occuring and assist in identification of autoantigens.
Reduction in TCR expression limits the response of myelin specific T cells (131.11)
Authors: Wasserman Heather Anong , Evavold Brian D 
Published: January 2023

Abstract
Previous studies have found that a 95% reduction in TCR expression does not adversely affect response to foreign antigen specific T cells. Since self-reactive T cells may have low affinity for self-antigen, we investigated whether myelin reactive T cells have lower affinity TCR for their specific ligand and are sensitive to a reduction in TCR expression. To test this concept, a MOG-specific Vα3.2 and Vβ11 TCR transgenic mouse model (referred to as 2D2 mice) is employed in our in vitro studies. Autoreactive MOG specific CD4+ T cells pre-incubated with Vα3.2 Ab have a dramatic dose-dependent reduction in proliferation. Quantitation of T cell receptor expression found that there are a maximum of 25,000 TCR per cell. A reduction of TCR expression by 31% led to a 50% reduction in proliferation, whereas a 68% TCR expression reduction correlates in a 67% reduction in proliferative response. Additionally, 2D2 cells cultured with Vα3.2 Ab and MOG 35-55, and then restimulated with MOG 35-55 also have a dose-dependent reduction in proliferation, suggesting that Ab treatment may lead to a possible phenotypic change resulting in T cell anergy. Therefore, TCR expression may provide a potential therapeutic window, which one can use to distinguish foreign antigen specific T cells from self-reactive T cells and allow for inhibition of autoimmunity without inducing global immune suppression.
This research was supported by NIAID (RO1 AI056017) and NMSS (RG 3683) grants
Expansion of GAD65–Specific Immunoregulatory Effector Cells by Biolistic-Mediated Gene Delivery Prevents Autoimmune Diabetes in NOD Mice (131.14)
Authors: Goudy Kevin , Li Li , Tisch Roland 
Published: January 2023

Abstract
Intramuscular (i.m.) vaccination with plasmid DNA (pDNA) encoding beta cell autoantigens such as glutamic acid decarboxylase 65 (GAD65) is an approach to suppress ongoing Type 1 diabetes (T1D) in NOD mice. However, protection is dependent on co-treatment with pDNAs encoding IL-4 and/or IL-10, and the subsequent induction of type 2 CD4+ T effectors. To enhance the therapeutic efficacy of pDNA vaccination, we explored epidermal delivery of pDNA via “gene gun”. Notably, reports have shown induction of a preferential type 2-like CD4+ T cell response when pDNA encoding antigen-only is delivered to the epidermis via gene gun. We hypothesized that biolistic delivery of pDNA would preferentially induce type 2 CD4+ T effectors and suppress established beta cell autoimmunity in NOD mice. Groups of 10 wk-old NOD female mice were treated four times with pDNA encoding GAD65-Ig (pGAD65) administered i.m. or delivered by gene gun on 1.6 uM gold particles. Whereas i.m. injection of pGAD65 resulted in preferential induction of GAD65-specific CD4+ Th1 cells and no protection against T1D, the majority of NOD mice treated with gene gun delivered pGAD65 remained diabetes-free and protection correlated with increased GAD65-specific IL-4 secreting CD4+ T cells. These results demonstrate that gene-gun delivered pDNA encoding beta cell autoantigen-only is an effective strategy to induce immunoregulatory CD4+ T cells and suppress T1D.