bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reduced acetylation of histones H3 in peripheral blood mononuclear cells of patients with Primary Sjögren’s syndrome
Authors: Li Yan 
Published: January 2023

Abstract
Sjögren’s syndrome (SS) is an autoimmune disease characterized by an exocrine glands epithelium injury with dense lymphocytic infiltrates, mainly composed of activated T and B cells, involvement of the peripheral nervous, pulmonary, blood, kidney, skin, and joint. Present at the interface of genetic and environmental risk factors, histone acetylation is suspected to play a key role in SS. To clarify this point, we investigated the expression of histone deacetylases (HDACs1–10) and histone acetyltransferase (HAT-p300 CREEBP PCAF) by real time PCR in peripheral blood mononuclear cells (PBMCs) from SS patients. Elisa measured HDACs\HATs and histone H3/ H4 acetylation activity. Flow Cytometry and western blot verified histone acetylation level. The expression of HDACs1-10, p300, CREEBP and PCAF in PBMCs from SS patients was decreased in mRNA comparison with healthy controls (HCs). Total histone acetylation was reduced in PBMCs from SS patients, while no difference was observed in histone deacetylation. The extent of total histone H3, but not H4, acetylation in PBMCs from SS patients was decreased compared to that in HCs. An altered balance between HDAC and HAT activity was found in SS PBMCs. As a consequence, part of the pathogenic mechanisms in SS may be linked to epigenetic modifications, thus opening new therapeutic perspectives in SS.
Peripheral blood neutrophil and G-CSF levels are linked to aztreonam lysine use in humans with cystic fibrosis
Authors: Wright Nathan Darrell , Gordon John Robert 
Published: January 2023

Abstract
The neutrophil response contributes substantially to disease progression in cystic fibrosis (CF). Research was directed at understanding the circulating neutrophil in CF. Medical chart data and 30 blood samples were collected from volunteers with CF (ΔF508/ΔF508; mean age 29 years) while 21 blood samples were collected from age-matched controls. Isolated plasma was then analysed for G-CSF and CXCL1 and statistically compared with neutrophil numbers and medical chart data, while signalling pathways in these neutrophils were investigated using kinome array approaches.
Our results indicated that the CF group being treated with the inhaled β-lactam antibiotic aztreonam lysine had significantly increased G-CSF levels (p&lt;.05) and isolated circulating neutrophil numbers (p&lt;.01) when compared to the CF group without the antibiotic. Neutrophil numbers were also significantly increased in both the inhaled aztreonam lysine positive (p&lt;.001) and negative (p&lt;.05) CF groups when compared to the non-CF control group. Neutrophil numbers in CF displayed a significant (p&lt;.05) positive correlation with G-CSF levels. We observed a significant (p&lt;.05) negative correlation between disease progression in CF (observed as FVC%) and neutrophil numbers. Kinome analyses indicated several altered pathways that warrant further investigation. These results revealed a connection in CF between antibiotic treatment, inflammatory markers in the blood, altered neutrophil signaling, changes in circulating neutrophil numbers, and disease progression. From a clinical perspective these results identified a potential connection between inhaled aztreonam lysine, circulating neutrophil numbers and decreased lung function in our CF volunteers.
Potential role of myeloid-derived suppressor cells in pulmonary fibrosis
Authors: Liu Tianju , Rinke Andrew , Flaherty Kevin , Phan Sem Hin 
Published: January 2023

Abstract
Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease without effective curative therapy. Recruitment of bone marrow-derived myeloid cells is implicated in lung fibrosis by promoting fibrosis via paracrine mechanisms. Recent evidence shows increased circulating myeloid-derived suppressor cells (MDSC) in IPF patients. More recently, the importance of MDSC is suggested by the observation of MDSC accumulation in IPF lungs. The objective of this study was to assess the accumulation/expansion of MDSC in bleomycin-induced mouse model of pulmonary fibrosis and human IPF, and investigate their potential role in lung myofibroblast differentiation. Flow cytometry analysis showed that MDSCs in lung were increased in a mouse model of lung fibrosis, as well as in peripheral blood samples from IPF patients. The increase of the monocytic subtype of MDSC (M-MDSC) was greater than that for the granulocytic subtype (G-MDSC) in the peripheral blood samples from patients with IPF. In vitro co-culture of normal mouse lung fibroblasts with sorted bone marrow (BM)-derived MDSCs using trans-well inserts revealed paracrine activation of lung fibroblasts by activated M-MDSC, but not GMDSC, as manifested by significantly elevated mRNA levels of α-smooth muscle actin and TGFβ mRNAs. Both quiescent M-MDSC and G-MDSC failed to stimulate fibroblast activation or myofibroblast differentiation. Interestingly, the level of TGFβ mRNA in M-MDSC was higher than G-MDSC. These findings indicated that in pulmonary fibrosis, BM-derived MDSCs were mobilized and recruited to the lung, and the recruited M-MDSC subpopulation subsequently played a paracrine role by activating resident lung fibroblasts and myofibroblast differentiation.
Simultaneous analysis of gene expression and T cell repertoire in peripheral blood mononuclear cells from patients with cystic fibrosis at the single cell level
Authors: Ferguson Annabel , Luo Jaidi , An Xiaojing , Pilewski Joseph M , Chen Wei , Chen Kong 
Published: January 2023

Abstract
Highly sensitive RNA sequencing (RNA-Seq) methods enables gene expression analysis of low-input samples and even single cells. Single-Cell RNA sequencing (scRNA-seq) is a powerful tool for studying the transcriptome and enables discovery of cellular differences usually masked by bulk sampling methods. The Chromium Single Cell Immune Profiling Solution developed by 10X genomics makes it possible to examine gene expression and T cell receptors simultaneously on a cell-by-cell basis. Patients with cystic fibrosis (CF) often suffer inflammatory lung diseases caused by chronic Pseudomonas aeruginosa (PA) colonization. This colonization could also skew the lymphocytes repertoire in peripheral blood mononuclear cells from CF patients. We investigated the use of scRNA-seq to assess the lymphocyte repertoire in CF patients with PA colonization and whether the single-cell transcriptomic changes correlate with preferential T cell receptor (TCR) usage. Our analysis did not find evidence for clonal expansion of T cells in peripheral blood of these CF patients, suggesting that PA colonization mainly elicits local adaptive immune responses in the lungs. However, this dataset did reveal a striking depletion of the population of T cells known as mucosal associated invariant T cells (MAIT cells) in CF patients. Within the MAIT cell population, we found an overrepresentation of certain genes that was observable in healthy but not CF samples. In this study we show the feasibility of simultaneous analysis of gene expression and lymphocytes repertoire in PBMCs. We recapitulate previous findings that implicate a protective role for MAIT cells against PA colonization, and also provide evidence for the involvement of particular genes in this effect.
The impact of IL-27 on human T cells is altered in multiple sclerosis patients
Authors: Clénet Marie-Laure , Laurent Cyril , Devergne Odile , Duquette Pierre , Prat Alexandre , Larochelle Catherine , Arbour Nathalie 
Published: January 2023

Abstract
Interleukin-27 (IL-27) exhibits pro- and anti-inflammatory properties. Although this cytokine dampens the severity of experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis (MS), whether IL 27 contributes to the pathobiology of MS remains unclear. We detected the IL-27 receptor (IL-27R) on brain-infiltrating T cells in MS tissues suggesting that they are susceptible to this cytokine. Notably, beneficial effects of MS immunomodulatory therapies are associated with IL-27 induction. We compared blood samples from untreated MS patients to healthy donors (HC) for i) serum levels of IL-27 and expression of IL-27R by T cells and ii) effects of IL-27 on phenotype and functions of T cells. Reduced proportions of CD4 T cells but greater percentages of CD8 T cells expressed IL-27R in MS patients compared to HC. IL-27 triggered phosphorylation of STAT1 (pSTAT1) and STAT3 (pSTAT3) in all T cell subsets; similar levels of pSTAT1 were induced in T cells from MS and HC. In contrast, increased proportions of CD4 and CD8 T cells exhibited pSTAT3 in response to IL-27 compared to cells from HC. Moreover, addition of IL-27 to activated T cells significantly decreased the percentages of GM-CSF or IL-17 producing CD4 and CD8 T cells from MS patients but had a minimal impact on HC’s cells. Finally, sera from MS patients contained elevated amounts of soluble IL-27 and IL-27Rα (a natural IL-27 antagonist) compared to HC. Our results reveal that IL-27 has an enhanced impact on T cells from MS patients, which could contribute to dampen key immune responses (e.g. GM-CSF, IL-17) associated with this inflammatory/autoimmune disease. However, elevated in vivo levels of its natural antagonist could hinder IL-27-mediated anti-inflammatory properties.
Morphometric and cytologic characterization of tertiary lymphoid follicles in resected bowel of patients with Diverticulitis, Ulcerative Colitis, and Crohn’s Disease
Authors: Mauner Aaron Laurence , McNamee Eoin N 
Published: January 2023

Abstract
The presentation of ectopic Tertiary lymphoid structures (TLS) in peripheral tissues is a pathological hallmark for a number of Autoimmune and Immune mediated conditions, not least within the bowel wall of patients with Crohn’s disease. However, there remains limited understanding of the cellular constituents or the function of TLS during intestinal pathology. To address this, we characterized the location, size, frequency and cellular constituents of B-cell follicles within resected bowel tissues from patients with Diverticulitis (DIV), Crohn’s Disease (CD) or Ulcerative Colitis (UC) (n=9–10/group). Samples were formalin fixed, paraffin embedded, serially sectioned, and subsequently stained by multiplexed immunohistochemistry. Morphometric analysis was performed with Olympus CellSens software and PerkinElmer Vectra 3.0 Automated Quantitative Pathology Imaging System and statistical analyses performed using R Statistical Software. Results show that the number, size and anatomic location of large B-cell aggregates differed greatly between disease states, with both UC and DIV follicles localizing primarily in the mucosa; while samples from patients with CD presented with mature Ki67+ TLS in the deeper layers of serosa and muscle. Multicolor IHC shows that TLS in Crohn's disease have large intra-germinal center CD4+ T-cell populations while UC samples contained a marked increase in both monocytes and neutrophils compared with both CD and DIV. These data suggest that despite ectopic TLS being present in a number of pathologic conditions, their composition within the bowel wall is disease state dependent and suggests altered germinal center function.
Monocytes of Chronic Fatigue Syndrome Patients, their Family Members without CFS, and Unrelated Healthy Donors: Searching for Differences
Authors: Hudig Dorothy , Tokunaga Keli , Sung Alexander P , Guglielmo Michael J , Smith-Gagen Julie , Merica Rebecca , Redelman Doug 
Published: January 2023

Abstract
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is characterized by fatigue that does not improve with rest and worsens with exertion. ME/CFS may involve inflammation in the brain and/or be associated with viral infections. Increased numbers of pro-inflammatory monocytes in blood are a hallmark of infection. Biomarkers for, and information about, the CFS disease process are greatly needed. Here we queried circulating monocytes. In a pilot study, CFS patients and family members without CFS from three families were compared in order to reduce genetic and environmental variables. The patients conformed to the Fukuda 1994 standards. Unrelated healthy control subjects matched to the patients were included in the study. Blood monocytes were stained and analyzed by flow cytometry: counted with TruCountR beads, or stained after isolation and overnight culture of isolated peripheral blood monocytes. M1 pro-inflammatory monocytes are CD16A negative, while anti-inflammatory monocytes are CD16A positive; CD2 is a marker for dendritic monocytes, and changes in forward scatter could indicate in vitro activation. Notably, there were statistically significant differences between the lower percentages of M1 monocytes of the CFS patients compared to the controls; however, these differences were not significant between the patients and their family members without CFS. Also, the CD16A MFIs of the patients' monocytes were lower compared to controls but not to family members. One conclusion is that it is important to include family members without CFS as a control group in the search for biomarkers to diagnose CFS.
CD4+ T cell antigen discovery in sarcoidosis
Authors: Greaves Sarah A , Mitchell Angela , Falta Michael , Santos Radleigh , Pinilla Clemencia A , Fontenot Andrew P 
Published: January 2023

Abstract
Sarcoidosis is an inflammatory granulomatous disease that primarily affects the lung. It has a worldwide distribution and affects individuals of all ages, races and both sexes. Despite the prevalence of the disease and continued investigation, the cause of sarcoidosis remains unknown. Evidence suggests that CD4+ T cells in the bronchoalveolar lavage (BAL) of sarcoidosis patients are instrumental in disease progression. Löfgren’s syndrome (LS) is an acute form of sarcoidosis that is accompanied by a specific set of inflammatory symptoms. In patients with LS, disease susceptibility has been associated with expression of HLA-DR3 and an expansion of BAL CD4+ T cells expressing T cell receptor (TCR) a-chain variable region (TRAV) 12-1. We hypothesize that in HLA-DR3+ LS patients, TRAV12-1 CD4+ T cell clones accumulate and expand in the lung in response to unknown antigens. To address the antigen specificity of these expanded CD4+ T cells, we performed single cell PCR of BAL CD4+ T cells derived from DR3+ LS patients to identify related abTCRs expressing TRAV12-1 and generated T cell hybridomas expressing these disease-relevant abTCRs. Using unbiased decapeptide positional scanning libraries, we have identified several stimulatory mimotopes. We used a biometrical analysis to obtain candidate etiologic peptides derived from human, viral, pollen, and mycobacterial protein databases and have identified several exogenous peptides that stimulate TCRs derived from the BAL of patients with active LS. Once etiologic antigens are validated, we will synthesize HLA-DR3-peptide tetramers to allow for more sophisticated approaches in the diagnosis and prognosis of this disease.
Antigenic Function of 14-3-3ζ is a Novel Tool to Treat Rheumatoid Arthritis
Authors: Chakravarti Ritu , McGowan Jenna , Peter Cara , Zhang Youjie , Veerman Brent , Rahaman Rafid , Joe Bina 
Published: January 2023

Abstract
Autoantigens are an integral part of autoimmunity, but their role in health and disease are mostly unknown. We identified 14-3-3ζ as an autoantigen in patients with thoracic aortic aneurysm caused by large vessel vasculitis. Like others, neither the cause nor the consequence of newly-identified antigenic function of 14-3-3ζ protein is known. To address this, we investigated the antigenicity of 14-3-3ζ by studying ex vivo effects on human peripheral blood mononuclear cells (PBMC) polarization, and in vivo using a rat autoimmune disease model. Exogenous 14-3-3ζ promoted PBMC proliferation and T cell polarization towards increasing Th1 and Th17 populations. A specific increase in Th1 cells and IFN-γ production provided evidence for MHC class II presentation of 14-3-3ζ antigen. These 14-3-3ζ-induced responses of T cell polarization and cytokine secretion were inhibited by immunosuppressive agents. Our in-depth studies further revealed that 14-3-3ζ-induced phosphorylation of MAPK led to NF-kB activation in macrophages and PBMC. In vivo administration of recombinant 14-3-3ζ in rats triggered strong innate and adaptive immune responses with significant increases in IL-17, IFN-γ, and anti-14-3-3ζ antibody. Importantly, the immunization of Lewis rats with 14-3-3ζ protected them from rheumatoid arthritis in Pristane-Induced-Arthritis (PIA) model. These collectively demonstrate that the antigenic 14-3-3ζ by acting as a DAMP promotes IFN-γ and IL-17 levels, and confers an anti-arthritic effect. To the best of our knowledge, this is the first report describing the antigenic function of 14-3-3ζ and its physiologically relevant anti-arthritic role.
IL-15 expression on B cells and myeloid cells is distinctly modulated by inflammatory factors in MS and healthy donors
Authors: Farzam-kia Negar , Moratalla Ana Carmena , Lemaître Florent , Beauseigle Diane , Haddad Elie , Larochelle Catherine , Prat Alexandre , Arbour Nathalie 
Published: January 2023

Abstract
Although the salience of the immune system in multiple sclerosis (MS) pathology has been established, the mechanisms involved are incompletely characterized. We have identified IL-15 as a relevant contributor to MS neuropathology; a greater proportion of antigen presenting cells (APC) (B cells and myeloid cells) from MS patients express IL-15 vs. controls. Our aim is to identify factors and mechanisms that contribute to elevated IL-15 levels in the context of MS. The addition of CpG, a TLR9 agonist, to CD40L-cultured human B cells increased the proportion of IL-15+ B cells. Whereas other TLR ligands (e.g. polyI:C) did not alter B cell IL-15 levels, they nevertheless increased CD40 expression. Monocytes from MS patients and age/sex-matched healthy controls (HC) were differentiated into M0 macrophages prior to M1 or M2 polarization. M2 polarization significantly enhanced the percentage of IL-15+macrophages; in contrast, M1 polarization did not. GM-CSF, which is produced by encephalitogenic T cells, induced a major increase in the proportion of IL-15+ M0 and M1 macrophages and decreased the proportion of IL-15+ monocytes from HC and MS donors. While IL-15+ M2 macrophages from HC are increased upon GM-CSF stimulation, IL-15+ M2 from MS donors are not. GM-CSF did not alter IL-15 expression on microglia. Pharmacological inhibitors of STAT5 (Pimozide) and JAK2 (AG490) phosphorylation decreased the GM-CSF-induced increase of IL-15 on M0 macrophages. Thus, GM-CSF is a potent trigger for IL-15 expression on human M0, M1 and M2 macrophages, but not monocytes or microglia. Our data suggest that specific stimuli mediate IL-15 expression on human APC and that cells from MS donors exhibit altered susceptibility to IL-15 modulators compared to HC.
NSG Mice Engrafted with Human PBMC Provide a Robust Platform for Preclinical Testing of Immunomodulatory Therapeutics Relevant to GvHD, Autoimmunity, and Oncology
Authors: Soper Brian , Rose Aaron , Hodel Ashley , Kaur Pali 
Published: January 2023

Abstract
NSG mice engraft human PBMC and allow efficacy testing of immunomodulatory drugs in a GvHD setting. Engraftment is progressive and at d5 PBL has 10% human cells with a CD4:CD8 T cell ratio of 3:1. The CD4 cells are 30% CD45RO− CCR7+ naïve (Tn), 40% CD45RO+ CCR7+ central memory (Tcm) and 10% CD45RO+ CCR7− effector memory (Tem). The CD8 cells at d5 contain 9% Tn, 3% Tcm, and 5% Tem. At d18 human cells are 70% with a CD4:CD8 ratio of 1:3, 0% Tn, 10% Tcm and are 22% CD4 Tem and 52% CD8 Tem. Engrafted mice were treated either prophylactically (day −1) or therapeutically (day 5) with Abatacept or an unknown test article. PBL at d10 post engraftment from mice treated at day −1 showed human cells at 14.6%, 2.2%, and 1.6% for vehicle, Abatacept, and test article, respectively. The Abatacept group were primarily CD4 Tn. Abatacept and test article treated mice showed 100% survival, almost no tissue infiltration, normal body weight, and were healthier based on disease activity index (DAI) scores. When dosed therapeutically, human cells at d18 were 67.3%, 16.9%, and 37.9%, respectively. Both treatments suppressed T cell expansion, maintained a 3:1 CD4:CD8 ratio and decreased development of the CD8 Tem pool (CD8 Tem 32.7%, 15.4%, and 4.9%, respectively). 100% of the d5 dosed Abatacept group survived, maintained body weight, and had improved DAI. 70% of the d5 dosed test article group survived and at d20 began losing body weight and accumulated poor DAI scores. The data presented here provide a detailed analysis of activated human T cell pools and progression of GvHD in NSG recipients of human PBMC. The model allows detailed analysis of in vivo modulation of T cell activity and therefore enables rapid evaluation of therapies relevant to not only GvHD, but to autoimmunity and oncology.
Detection and Characterization of T cells Reactive to Hybrid Insulin Peptides in Patients with Type 1 Diabetes
Authors: Baker Rocky Lee , Gottlieb Peter A , Delong Thomas , Haskins Kathryn 
Published: January 2023

Abstract
Our lab has recently established that Hybrid Insulin Peptides (HIPs) are antigenic ligands for diabetogenic T cell clones in the NOD mouse model of autoimmune diabetes. Furthermore, these peptides are detected in the islets of NOD mice, indicating that islet-infiltrating T cells encounter their antigen at the site of immune injury. Importantly, we have found that the phenotype of HIP-reactive T cells present in the peripheral blood of NOD mice is indicative of ongoing autoimmunity in the pancreas. In the human setting, we demonstrated that T cells reactive to HIPs are present in the residual islets of type 1 diabetic (T1D) organ donors. In this study, using an IFN-g ELISPOT assay, we investigated whether T cells reactive to a panel of 16 HIPs could be observed in the PBMCs of T1D patients or control subjects. We observed that about a third of the T1D patients responded to at least one HIP; 70% of the responding patients (but only one control subject) showed reactivity to multiple HIPs. Responses to 4 HIPs were significantly elevated in T1D patients but not in control subjects. Using a multiplexed cytokine assay, we determined that T cells from responder patients had a bias towards a Th1 phenotype upon stimulation with HIPs. We isolated 8 non-redundant, antigen-specific T cell clones reactive to 6 different HIPs. These T cell clones react to their target HIPs in the low nanomolar range and secrete IFN-g, TNF-a and GM-CSF. This is the first report that HIP-reactive T cells with an inflammatory phenotype can be detected in the peripheral blood of patients with T1D, indicating that the presence of HIP-reactive T cells might serve as a biomarker of disease activity in the human setting.