bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combinatorial ex vivo analysis of beta cell specific CD4+ T-cell responses reveals a predominance of proinsulin specific cells
Authors: Blahnik-Fagan Gabriele , Uchtenhagen Hannes , Chow I-Ting , Kwok William W , James Eddie A 
Published: January 2023

Abstract
CD4+ T cell responses are thought to play a key role in the progression of type 1 diabetes (T1D). However, effective monitoring and characterization of T cells that respond to beta cell epitopes in subjects with T1D has been limited by technical obstacles, including the inherently low frequencies of autoreactive CD4+ T cells and the variable responsiveness of individual subjects to single epitopes. We have recently implemented a combinatorial staining approach that allows direct ex vivo characterization of multiple CD4+ T-cell specificities in a single sample. We applied this combinatorial HLA class II multimer assay to directly measure the frequency and phenotype of beta cell specific CD4+ T cells. For this work we utilized five peptides that were previously identified as naturally processed DRB1*04:01 restricted epitopes from proinsulin, GAD65, IA-2, and IGRP. Although responses to each of these peptides can be readily detected after in vitro expansion culture, our results indicated that only proinsulin specific T cells were consistently present at high frequencies in subjects with T1D. Phenotypic analysis of beta cell specific CD4+ T cells revealed that subjects with T1D had a higher frequency of proinsulin specific T cells with a memory phenotype than HLA matched controls. The majority of these memory cells were CXCR3 positive and an increased percentage were CCR7 negative, suggesting that Th1-like effector memory responses are present in subjects with T1D. Finally, this approach is compatible with combinatorial class I multimer analysis, facilitating the characterization of self-reactive CD4+ and CD8+ T cells using a single sample.
T Cells derived from Peripheral lymph nodes have different activation requirements than T cells isolated from spleen.
Authors: Elhofy Aelhofy 
Published: January 2023

Abstract
Primary murine T cells have been used as a model for human immune function. Animal models like EAE have shown disparate activities between splenic T cells and T cells derived from peripheral lymph nodes (PLN T cells). In this study we investigated if PLN T cells had the same activation requirements as splenic T cells in a serum free environment. PLN T cells activated in did not proliferate or secrete cytokines serum free conditions either by the non-specific stimuli of anti-CD3 and anti-CD28, or by PLP139-151 specific antigen re-stimulation. Conversely, splenic T cells cultured in serum free or FBS conditions proliferated and secreted cytokines equally in both activation models. The activation of serum free PLN T cells can be rescued with the addition of serum albumin as proliferation and cytokine production in both activation models was equivalent to those cultured in FBS. The activation requirement was specific to serum albumin, as the addition of other proteins including ovalbumin and serum proteins; gamma globulin, and collagen did not result in PLN T cell activation. Furthermore full antigen specific activity was returned to the PLN T cells with the addition of serum albumin demonstrated by the ability to induce adoptive transfer of EAE. The use of FBS in culture has masked the differential activation requirement of T cells depending on where they were derived and this studied showed serum albumin is requisite for the activation of T cells derived from PLN but not from spleen.
Dual T cell receptor-expressing CD8 T cells potentiate autoreactivity
Authors: Tsai Kevin , Huang Yu-Hsuan , Wang Xiaoxia , Priatel John J 
Published: January 2023

Abstract
Central tolerance serves to eliminate newly developing T cells that express strongly autoreactive T cell receptors. Although central tolerance is efficient in deleting high avidity autoreactive T cells, some lower avidity autoreactive T cells escape negative selection to cause autoimmune diseases. Although tight allelic exclusion limits thymocytes to expressing a single TCRbeta chain, rearrangement of the TCRalpha chain continues unabated until halted by positive selection, enabling thymocytes to express up to two TCRalpha chains and thus two TCRs. Moreover, it has been postulated that pathogenic low avidity autoreactive CD8 T cells may escape central tolerance through expression of a secondary benign TCR that mediates positive selection. To determine the role of dual TCR expressing CD8 T cells in autoreactivity and autoimmunity, we have compared CD8 T cell autoreactivity against the model autoantigen ovalbumin between T cells capable of expressing two TCRs (TCRalpha+/+) versus T cells capable of expressing a single TCR (TCRalpha+/−). TCRalpha+/− CD8 T cells exhibited reduced proliferative capacity upon OVA stimulation relative to TCRalpha+/+ CD8 T cells. In addition, a lower frequency of TCRalpha+/− CD8 T cell effectors produced IFN-gamma upon activation with OVA compared to TCRalpha+/+ CD8 T cell effectors. Taken together, we shown that dual TCR expression by CD8 T cells reduces the efficiency of T cell tolerance and may potentiate T cell autoreactivity. We are investigating whether dual TCR-expressing T cells are key to the pathogenesis of autoimmune diabetes in our mouse model system. Our results will provide valuable insight into the escape mechanisms exploited by pathogenic autoreactive CD8 T cells.
Type I interferon signaling is essential for cytotoxic T lymphocytes to infiltrate into the pancreatic islet and cause type I diabetes
Authors: Marro Brett S. , Ware Brian C. , Oldstone Michael B.A. 
Published: January 2023

Abstract
Type I diabetes (T1D) is characterized by elevated blood glucose, low pancreatic insulin and lymphocyte migration into Islets of Langerhans with destruction of insulin-producing β-cells. A seminal question in the pathogenesis of T1D is the signal(s) that control lymphocyte entry and attack on insulin-producing β-cells. Numerous epidemiological studies in humans suggest that infectious agents may initiate T1D. To model T1D in the mouse and probe the underlying mechanisms governing disease, several years ago we engineered Tg mice to solely express the glycoprotein gene of Lymphocytic Choriomeningitis Virus within β-cells using the rat insulin promoter (RIP-LCMV-Gp Tg mice). In this Tg model, infection with LCMV generates Gp-specific CD8+ CTL that target β-cells, leading to T1D by day 14–20 post-infection. We now report that blockade of Type I interferon signaling via antibody-mediated neutralization of the Type I interferon receptor (IFNAR) or its cognant ligands interferon-α (IFNα) or interferon-β (IFNβ) suppress the selective infiltration of virus (autoimmune) specific CD8+ T cells into the islet, thereby sparing islets from destruction and preventing T1D. During this early phase of viral challenge (up to day 20), neutralization of either IFNα or IFNβ did not affect the expansion of virus (autoimmune) specific CD8+ T cells or impact viral clearance, but nevertheless abrogated disease by restricting CD8+ T cells to the rim of the islets, thus preventing access of T cells into the islet. These results demonstrate that IFNα and IFNβ are central orchestrators in developing diabetes by allowing the entry of lymphoid cells into the islet that then target and kill insulin-producing β-cells.
The pathogenic effect of TNF-α on development of Sjögren’s syndrome in non-obese diabetic mice
Authors: Zhou Jing , Yu Qing 
Published: January 2023

Abstract
Tumor necrosis factor-alpha (TNF-α), is a proinflammatory cytokine involved in the pathogenesis of various autoimmune diseases including rheumatoid arthristis, inflammatory bowel disease and psoriasis. Previous studies showed that patients with Sjögren’s syndrome(SS), a prevalent systemic autoimmune disease primarily affecting tear and saliva secretion, exhibit enhanced expression of TNF-α in saliva and salivary gland. However, the exact role of TNF-α in SS disease development is not yet fully determined. Our current study employed non-obese diabetic (NOD) mice, a model of SS-like diseases, to investigate the function of endogenous TNF-α in pathogenesis of SS. We observed that administration of a neutralizing anti-TNF-α antibody significantly improved salivary secretion in female NOD mice, indicating a remission of clinical symptom of SS. TNF-α neutralization also decreased the number of leukocytic foci and reduced the number of total and activated CD4 T cells and B cells in the submandibular gland of NOD mice. Moreover, neutralization of TNF-α reduced the number of CXCR3-expressing CD4 T cells and T-bet protein level in the submandibular gland, suggesting a decrease in the infiltrating T helper 1 and T cytotoxic 1 cells. In addition, the decrease in CXCR3+ CD4 T cells was associated with reduced expression of CXCR3 ligand CXCL9. Overall, the results presented here indicate that endogenous TNF-α plays a pathogenic role in the initiation and development of SS disease and neutralization of TNF-α may be greatly beneficial for the prevention and treatment of this prevalent autoimmune disease.
S1P1 deletion differentially affects TH17 and Regulatory T cells
Authors: Eken Ahmet , Duhen Rebekka , Singh Akhilesh K , Fry Mallory , Buckner Jane H , Kita Mariko , Bettelli Estelle , Oukka Mohamed 
Published: January 2023

Abstract
Sphingosine-1 phosphate receptor 1 (S1P1) is a G-protein coupled receptor critical for the egress of T and B cells out of lymphoid organs. Although S1P1 agonists (such as fingolimod) are currently used for the treatment of multiple sclerosis (MS) little is known how S1P1 signaling regulates Th17 and Treg cell homeostasis. To study the impact of S1P1 signaling on Th17 and Treg biology, we specifically deleted S1P1 in Th17 and Treg cells using IL-17ACre and Foxp3Cremice, respectively. Deletion of S1P1 in Th17 cells conferred resistance to experimental autoimmune encephalomyelitis (EAE) characterized by reduced Th17 cell distribution across peripheral organs and diminished Th17 cell generation. On the other hand, permanent deletion of S1P1 in Treg cells resulted in autoimmunity and acute deletion rendered mice more susceptible to EAE. Importantly, our study revealed that S1P1 not only regulated the egress of Treg cells out of lymphoid organs and subsequent non-lymphoid tissue distribution but also their phenotypic diversity. Most of the Treg cells found in S1P1-deficient mice had an activated phenotype and were more prone to apoptosis, thus converted to effector Treg. The comparison of Treg cells obtained from MS patients treated with fingolimod to those treated with other oral drugs confirmed the switch of Treg cells into effector memory phenotype. Our results provide novel insight into the functions of S1P1 and potential impact of long term fingolimod use on Th17 and Treg cell biology and general health in MS patients.
CNS tissue response to Multiple Sclerosis
Authors: Berl Sabina , Waisman Ari 
Published: January 2023

Abstract
A well known inflammatory, autoimmune disease of the central nervous system (CNS) is multiple sclerosis (MS), where cells of the immune system invade the CNS, causing demyelination and neurodegeneration and leading to paralysis. The most common animal model of MS is experimental autoimmune encephalomyelitis (EAE). Not much is known about the response of neurons to the inflammatory triggers, although damage to them is the main initiator for paralysis in MS patients and EAE, due to the difficulty to separate and analyze them for gene expression on single cell level.
The aim of this project is to isolate and analyze neuronal mRNA expression level from adult mice. Isolated neuronal mRNA of naïve mice and mice with EAE will be sequenced by Next Generation Sequencing (NGS) to better understand how neurons respond to damage induced by inflammation.
RiboTag mice have a modified ribosomal protein L22 including a HA-tag. After crossing the RiboTag mice to the neuron-specific NFH-Cre mice, HA-tagged ribosomes will be specifically expressed in neurons giving the possibility to immunoprecipitate only neuronal RNA.
Immunoprecipitated neuronal mRNA of the RiboTag mice will be further analyzed by deep sequencing using NGS technology.
The final goal of this study is to analyze gene expression pattern of neurons that are attacked by pro-inflammatory cells and mediators during EAE. Understanding how neurons respond during autoimmune disease should allow to design better therapies that will protect these cells under such pathogenic conditions. This work has implications not only for patients of MS, but also for other neurological diseases, including Alzheimer’s disease and stroke.
Mycoplasma superantigen enhances the severity of K/BxN autoimmune arthritis through TLR4/BCL6/TFH pathway
Authors: Mu Hong-Hua , Nourian Maziar M. , McKennon Kevin B , Wang Jingyi 
Published: January 2023

Abstract
Mycoplasma arthritidis mitogen (MAM), a potent superantigen (SAg) secreted by M. arthritidis, is known to contribute to mouse inflammatory/autoimmune arthritis. We previously demonstrated that C3H mouse strains differing in expression of TLR4, also differ in Th-1 versus Th-2 and Th17 cytokine profiles in response to MAM due to a differential engagement of TLR2 and TLR4. HMGB-1, a member of heat shock protein family that is abundantly expressed in many cell types upon activation, has been implicated in regulating TLR signaling pathways following its interaction with RAGE. In the present study, we established a new model of mouse arthritis induced by injection of K/BxN sera. Also, we provide evidence that HMGB1 is an essential endogenous activator of the immune response in K/BxN-sera transfer BALB/c mice following MAM immunization. Level of HMGB1 expression was promptly increased in K/BxN sera-transferred mice after MAM injection. The increase in HMGB1 was required for initiation of MAM-induced enhancement of BCL-6, type I interferon (IFN) and follicular helper T cell responses. Blockade of HMGB1 caused prolonged activation of BCL-6 pathways, reduced level of type I IFN and augmented production of IL-10. Also, MAM-immunized K/BxN mice injected with HMGB1 antibody were less susceptible to arthritis suggesting HMGB1 may accomplish this mainly through its interaction with TLR4. Thus, HMGB1, in synergy with mycoplasma superantigen MAM, functions as biologically important modulators of TLR4/BCL6/TFH, and thus the targeting of this pathway can be a useful therapeutic strategy for treating human rheumatoid arthritis.
Mechanisms of Coagulation-Complement pathway interplay in ischemia/reperfusion induced tissue injury.
Authors: Robbins Racheal , Peng Suzette , Moratz Chantal M 
Published: January 2023

Abstract
Impaired fibrinolysis in systemic lupus erythematosus (SLE) is a contributing factor in disease complications including hyper-coagulation, tissue ischemia/reperfusion (IR), and increased risk of thrombosis: including stroke and deep vein thrombosis. Aberrant complement activation has been identified as a major factor in tissue injury in SLE. Previous work in the laboratory focused on mechanisms of complement activation in IR induced tissue injury in B6.MRL/lpr mice, an autoimmune prone mouse strain with a propensity to develop lymphadenopathy, glomerulonephritis, and polyarthritis. This work identified a C3-independent mechanism of downstream complement C5a generation contributing to tissue injury. Evidence of interplay between the coagulation and complement pathways was identified. Current work elucidates the mechanism by which the two pathways interact. Using a superior mesenteric artery IR model, we utilized complement and specific extrinsic or intrinsic coagulation pathway inhibitors to evaluate and compare resulting tissue pathology, generation of secondary inflammatory mediators, extent of innate immune activation, and generation of breakdown products involved in regulating feedback regulatory loops after ischemic/reperfusion injury. Inhibition of the extrinsic pathway had a greater impact on downstream complement activation, tissue pathology, vascular integrity, and recruitment of innate immune components versus the inhibition of the extrinsic pathway. Understanding the mechanisms of interplay between the pathways in the autoimmune mouse has significant clinical implications – especially in autoimmune diseases in which inflammation and hypercoagulability are known manifestations.
Intestinal Bacterial Composition of Lymphopenic Mice and Susceptibility to CD45RBhigh T Cell-Induced Colitis
Authors: Webb Cynthia Vannessa Reinoso , Koboziev Iurii , Furr Kathryn L , Phillips Caleb , Kottapalli Kameswara Rao , Ostanin Dmitry V , den Bakker Hendrik C , Grisham Matthew B 
Published: January 2023

Abstract

Background
Following relocation to our current institution we observed a 40% reduction in the incidence of T cell-induced colitis in our well-established mouse model of IBD. The objectives of this study were to: a) quantify and compare the colonic microbiota in healthy and colitic mice obtained from our current (TTUHSC) and former insitution (LSUHSC), and b) determine whether colonization of healthy mice housed at our current institution with feces from healthy or colitic mice from LSUHSC alters the incidence and/or severity of colitis.

Methods
DNA from frozen feces was isolated using standard protocols and 16S rRNA sequencing was performed using the Illumina MiSeq platform. For some studies, RAG1−/−mice were colonized (via gastric gavage) with 40 mg donor feces 1 week prior to adoptive transfer of naïve (CD4+CD45RBhigh) T-cells.

Results
Intestinal microbial populations are markedly different between the two institutions in healthy and colitic animals. Transplant of fecal microbiota from LSUHSC mice with chronic colitis into healthy RAG−/− recipients accelerates the onset, increases the incidence (&gt;95%) and enhances the severity of chronic colitis following T cell transfer. However, colonization of healthy WT or RAG−/− mice (in the absence of T cell transfer) with colitic feces did not induce disease over the 8 week observation period.

Conclusion
We conclude that colonization of healthy RAG−/− mice with dysbiotic but nonpathogenic microbiota obtained from colitic mice markedly increases the incidence and severity of chronic colitis in the T cell transfer model.
Characterization of the inflammatory effects of λ-carrageenan at the genomic and proteomic levels
Authors: Lewandowski Annie , Deaver Christine , Myers Michael J 
Published: January 2023

Abstract
λ-Carrageenan (λCGN) is often used to model local inflammation, but the mechanism by which this compound functions at the genomic and proteomic level has not been delineated. Whole blood from swine was stimulated with λCGN to determine inflammatory biomarkers. We also utilized a HEK-293 (human embryonic kidney) cell line transfected with a SEAP (secreted embryonic alkaline phosphatase)-release plasmid to determine which TLR (Toll-Like Receptor; TLR) signaling cascade was stimulated. Results indicated that Monocyte Chemotactic Protein-1 gene expression and protein were increased by λCGN stimulation while Serum Amyloid A gene expression and protein remained unchanged. λCGN stimulation increased mRNA levels of Phosphatidylinositol 3-Kinase and Alveolar Macrophage Chemotactic –II, decreased mRNA levels of Thrombospondin-2 Precursor, and had no effect on Interleukin-6 mRNA levels. λCGN stimulation increased protein levels of Interleukin-8 and had no effect on protein levels of Tumor Necrosis Factor-α. SEAP release from HEK-293 cells stimulated with λCGN indicated that both TLR2 and TLR4 signaling cascades were initiated. λCGN stimulation did not activate TLR3, TLR5, TLR7, TLR8, TLR9, NOD1, NOD2, Dectin-1a, or Dectin-1b. Experiments silencing either TLR1 or TLR6 to determine which TLR2 heterodimer is activated by λCGN are currently underway. In conclusion, while λCGN stimulation appears to signal through both the TLR2 and TLR4 pathways, significant differences exist between the genes and proteins stimulated by λCGN and pure TLR4 agonists such as E. coli-derived lipopolysaccharide.
Ron receptor tyrosine kinase plays a protective role in diet induced obesity and associated steatohepatitis
Authors: Allen Joselyn Natasha , Zhang Limin , Yu Shan , Balandaram Gayathri , Peters Jeffrey M , Patterson Andrew D , Hankey Pamela A 
Published: January 2023

Abstract
Obesity is one of the leading causes of preventable death in the U.S and a well-established risk factor for several pathological conditions including steatohepatitis. The activation of inflammatory tissue resident macrophages has proven to play a key role in the progression of the metabolic syndrome. Consequently, mechanisms that manipulate macrophage phenotype has become of great interest for developing therapeutic strategies for combating obesity and its associated diseases. Our lab has shown that Ron tyrosine kinase, a receptor expressed on tissue resident macrophages, limits an inflammatory response while promoting tissue repair. In this study, we first investigate the impact of Ron signaling on metabolic features preceding steatohepatitis using high fat diet (HFD) fed wild type (WT) and Ron deficient (Ron KO) mice. Ron KO mice demonstrated more severe events associated with obesity such as increased adiposity, exacerbated hyperglycemia and trend toward increased liver weights when compared to WT animals. Altered Ron signaling resulted in a shift in the plasma lipoprotein distribution from primarily VLDL and LDL to predominantly HDL in the WT control mice. To then establish the role of Ron signaling in steatohepatitis, high cholesterol diet (HCD) fed ApoE KO and Ron KO ApoE KO mice (DKO) were evaluated. We showed that DKO mice experience more severe steatohepatitis which is attributed to an increased prevalence of activated inflammatory Kupffer cells. In vitro studies ascertain that Ron manipulation of macrophage phenotype is in part, a result of downstream AMPK activation. These data indicate the role of Ron in attenuating obesity and its associated diseases and identify a novel downstream pathway involved in this process.
Localization and trafficking of pulmonary dendritic cells during early stages of airway inflammation
Authors: Lyons-Cohen Miranda R , Nakano Hideki , Whitehead Gregory S , Cook Donald N 
Published: January 2023

Abstract
Pulmonary dendritic cells (DCs) take up inhaled antigens, migrate to draining lymph nodes (LNs), and stimulate antigen-specific T cells. Bona fide pulmonary DCs (CD11c+CD88−) comprise CD103+ and CD11b/CD172a+ subsets, and their migration from lung to LNs is directed by the chemokine receptor, CCR7. However, the precise localization of these two different DC subsets in the lung, and how they move during the early stage of inflammation, are largely unknown. In this study, using multi-color high resolution imaging of precision cut lung slices, we found that at steady state, both CD103+ and CD172a+ DCs reside immediately adjacent to airways lined with CD324/E-cadherin-expressing epithelial cells. Interestingly, this region of the airways is also closely associated with lymphatic vessels, whereas regions lacking CD324 are not associated with either DCs or lymphatic vessels. Inhalation of LPS induced a Ccr7-gfp reporter gene within 2 h post-exposure, but the DCs remained closely associated with airways. By 6 h post- exposure, the number of airway-associated DCs decreased, while DCs associated with lymphatic vessels significantly increased. Very few DCs were detected in lung parenchyma. These observations suggest that after sensing danger signals, DCs translocate directly from the airway to lymphatics without first transiting through the parenchyma. In agreement with this rapid migration, activation of antigen-specific T cells was detected in regional LNs within 6 h of exposure.