bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quaternary Ammonium Compound Disinfectants Reduce Lupus-Associated Splenomegaly by Targeting Neutrophil Migration and T-Cell Fate
Authors: Abdelhamid Leila , Cabana-Puig Xavier , Mu Qinghui , Moarefian Maryam , Swartwout Brianna , Eden Kristin , Das Prerna , Seguin Ryan P. , Xu Libin , Lowen Sarah , Lavani Mital , Hrubec Terry C. , Jones Caroline N , Luo Xin M. 
Published: February 2023

Abstract
Hypersensitivity reactions and immune dysregulation have been reported with the use of quaternary ammonium compound disinfectants (QACs). We hypothesized that QAC exposure would exacerbate autoimmunity associated with systemic lupus erythematosus (lupus). Surprisingly, however, we found that compared to QAC-free mice, ambient exposure of lupus-prone mice to QACs led to smaller spleens with no change in circulating autoantibodies or the severity of glomerulonephritis. This suggests that QACs may have immunosuppressive effects on lupus. Using a microfluidic device, we showed that ambient exposure to QACs reduced directional migration of bone marrow-derived neutrophils toward an inflammatory chemoattractant ex vivo. Consistent with this, we found decreased infiltration of neutrophils into the spleen. While bone marrow-derived neutrophils appeared to exhibit a pro-inflammatory profile, upregulated expression of PD-L1 was observed on neutrophils that infiltrated the spleen, which in turn interacted with PD-1 on T cells and modulated their fate. Specifically, QAC exposure hindered activation of splenic T cells and increased apoptosis of effector T-cell populations. Collectively, these results suggest that ambient QAC exposure decreases lupus-associated splenomegaly likely through neutrophil-mediated toning of T-cell activation and/or apoptosis. However, our findings also indicate that even ambient exposure could alter immune cell phenotypes, functions, and their fate. Further investigations on how QACs affect immunity under steady-state conditions are warranted.
Interferon alpha promotes extrinsic apoptosis of keratinocytes following exposure to ultraviolet B radiation.
Authors: Estadt Shannon N , Moore Tyson M. , Kahlenberg J Michelle 
Published: February 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by sensitivity to ultraviolet radiation (UVR). UVR triggers cutaneous and systemic disease flares, yet mechanisms driving this are not well characterized. Type I interferons (IFNs) are overexpressed in non lesional SLE skin and promote death of SLE keratinocytes (KCs) after UVR, but the manner in which they do so is not known. This study explores activation and regulation of cell death pathways in KCs exposed to type I IFNs and UVR. We treated immortalized human KCs (N/TERTs) overnight with IFNα prior to UVB exposure. Four hours post UVB, IFNα priming significantly increased the percentage of cleaved-caspase-3+ (CC3+) and Annexin V+ propidium iodide− (AV+PI−) cells compared to treatment with UVB alone. Further, IFNα promoted apoptosis at a lower dose of UVB. Inhibiting RIP1, RIP3, or caspase-1 had no effect on death suggesting no involvement of necroptosis or pyroptosis. Use of a pan-caspase inhibitor or a caspase-8 inhibitor significantly reduced AV+PI− cells following IFNα and UVB treatment while caspase-9 inhibition did not, suggesting enhanced activation of extrinsic apoptosis. Treatment with IFNα increased expression of the death ligand TRAIL, an inducer of extrinsic apoptosis. Further studies will determine if IFN-induced upregulation of TRAIL is responsible for UVB-enhanced apoptosis in an autocrine fashion. Together, these data suggest that photosensitive responses exhibited by SLE patients may be due to type I IFN priming of KCs that sensitizes cells to undergo increased extrinsic apoptosis after minimal exposures to UVB. Continued investigation into mechanisms by which this occurs will provide important prophylactic options to prevent SLE flares.
NOX2-deficient neutrophils facilitate joint inflammation in immune-mediated arthritis
Authors: Liao Yi-Chu , Chen Peng-Chieh , Shieh Chi-Chang K 
Published: February 2023

Abstract
Immune-mediated inflammatory arthritis, including rheumatoid arthritis (RA), is characterized chronic inflammatory in the joints. NADPH oxidases 2 (NOX2) complex in phagocytes mediates the generation of high concentrations of reactive oxygen species (ROS) to kill the engulfed microbes. Inherited mutations in the subunits of NOX2 result in chronic granulomatous disease (CGD). CGD patients experience recurrent infections and diverse autoimmune conditions including arthritis. In our previous studies, we found that NOX2-deficient mice tended to have more severe inflammatory arthritis and neutrophils accumulation. Based on the hypothesis that NOX2-deficient neutrophils play a role in facilitating immune-mediated arthritis, we investigated the activity of neutrophil from inflamed joints. We found that while neutrophils from both NOX2-deficient and WT mice weakened the anti-CD3 and anti-CD28 induced T lymphocyte responses, NOX2-deficient neutrophils were less suppressive. Down-regulated PD-L1 expression on NOX2-deficient neutrophils was observed, which could be a cause for their impaired suppressive function. Moreover, pro-inflammatory genes, including IL-1b, NOS2, and Cxcl3 were up-regulated in NOX2-deficient neutrophils according to our RNA-seq results. In humans, neutrophils from RA joints were found to suppress the T cell proliferation. Consistent with our mice model, carbonylated protein and total GSH level, the oxidative markers, were lower in synovial fluid of RA patients. Our results implicate that neutrophil-mediated immune regulation may be an important factor in controlling inflammation in the joints, and this regulation is affected by the NOX2-produced ROS.
Loss of the TAM receptor Mer contributes to Sjögren’s syndrome-like disease in mice
Authors: Witas Richard M , Nguyen Cuong Q 
Published: February 2023

Abstract
Sjögren’s syndrome (SjS) is a chronic autoimmune disease primarily involving the exocrine glands where the involvement of the innate immune system is largely uncharacterized. Mer signaling has been found to be protective in several autoimmune diseases but remains unstudied in SjS. Here we report that soluble Mer (sMer) levels were elevated in SjS patient sera and positively correlated with focus score, ocular staining scores, rheumatoid factor, and Ro60 autoantibody levels. Increased sMer was also detected in C57BL/6.NOD-Aec1Aec2 (SjSS) mouse sera, coinciding with higher ADAM17 activity, the enzyme responsible for cleavage of Mer, and inactivation of such. Mer signaling outcomes were observed to be diminished in SjSS mice as evidenced by impaired efferocytosis in SjSS mouse macrophages and decreased Socs1 and Socs3 expression in SjSS salivary gland (SG). SjS diagnostic criteria were examined in MerKO mice to evaluate the protective role of Mer in SjS disease pathogenesis. MerKO mice developed SG infiltrates of B and T lymphocytes, SG apoptotic cells, antinuclear antibodies, and reduced saliva flow. Together, our data suggest a sequence of events where enhanced ADAM17 activity increases Mer inactivation, depresses Mer signaling, and removes a protection against the loss of self-tolerance and onset of autoimmune disease in SjSS mice.
Inflammation targets CDK11 in autoimmune diabetes as a protective mechanism against beta cell apoptosis.
Authors: Mora Conchi , Sala Ester , Vived Celia , Luna Júlia , Saavedra-Ávila Noemí-Alejandra , Sengupta Upasana , Haba Laura , Pizarro Javier , Verdaguer Joan , Stratmann Thomas , Vázquez-Carrera Manuel , Lahti Jill M 
Published: February 2023

Abstract
Autoimmune diabetes (T1D) is caused by the immune-mediated destruction of insulin-producing beta cells in the pancreas leading to hyperglycemia. Apoptosis is the main mechanism responsible for beta cell demise, including Fas-receptor engagement and Perforin release as essential death executers. However, the chronic inflammatory milieu pervading pancreatic islets prior to diabetes onset, alters beta cells function and primes beta cells for apoptosis long before the debut of the disease. To identify those genes that are targeted by inflammation and are causally related to beta cell fitness and viability we have used the microarray technology. We have identified several candidate genes being downregulated by inflammation, one of this genes is Cdk11, a cyclin-dependent kinase which is involved in transcription (CDK11p130), mitosis and apoptosis (CDK11p58). L-type cyclins are the partners for Cdk11p130, and Cyclin D3 is the partner of CDK11p58. The Cdk11 gene, when is transcribed, generates a single mRNA form, that can be translated either into CDK11p130 or CDK11p58 by a differential IRES (Internal Ribosome Entry Site) usage, being CDK11p58 only expressed during mitosis, while CDK11p130is expressed during all cell cycle phases. We have addressed the role of CDK11 in autoimmune diabetes using the T1D-prone NOD mouse model hemideficient in CDK11, since the CDK11 null mutation is embryonically lethal. First of all, we have confirmed the effect of inflammation on Cdk11 mRNA expression in islet cells by submitting NODSCID mice to adoptive transfer of increasing amounts of diabetogenic NOD splenocytes, and, observed that Cdk11 expression in islet endocrine cells is downregulated by inflammation in a dose-response dependent manner.
Vitamin A Deficiency Deteriorates Lupus Nephritis.
Authors: Abdelhamid Leila , Cabana-Puig Xavier , Swartwout Brianna , Zhu Jing , Li Yaqi , Ross A. Catharine , Cecere Thomas E. , Ruiz Phillip , Reilly Christopher M. , Luo Xin M 
Published: February 2023

Abstract
Vitamin A deficiency (VAD) is observed in both human and mice with lupus nephritis (LN). However, whether VAD is a driving factor for LN progression is unclear. Here, we investigated the effect of VAD on the progression of LN in a lupus-prone mouse model, MRL/lpr. In these mice, while LN is not apparent before 8 weeks of age, the inflammation of lymphoid tissues suggesting initiation of lupus is evident at weaning. Thus, we initiated VAD either before or after weaning to reveal a potential time-dependent effect. At around 15 weeks of age with both types of VAD, we found exacerbated LN that was characterized by deteriorated kidney inflammation, impaired renal epithelial integrity, and dramatic squamous metaplasia of the renal pelvic urothelium. Both types of VAD provoked severe neutrophilic tubulointerstitial nephritis and accelerated renal failure. This was concomitant with significantly higher mortality in all VAD mice. Looking at an earlier time point of 7 weeks of age and before the onset of clinical LN, both types of VAD increased splenomegaly, lymphadenopathy, and circulating autoantibodies; three additional hallmarks of lupus, as well as renal infiltration of conventional and plasmacytoid dendritic cells. These results suggest VAD-driven deterioration of LN regardless of the time of VAD initiation. Our findings raise significant public health concerns that concurrent lupus and VAD may lead to more severe SLE. Future studies will elucidate the underlying mechanisms of how VAD modulates myeloid cells to accelerate the initiation of LN.
Specific traits of the B cell response to self-antigens revealed by single cell transcriptomics
Authors: Castrillon Carlos Eduardo , Simoni Lea , van den Broek Theo , van der Poel Cees , Akama-Garren Elliot H , Carroll Michael C 
Published: February 2023

Abstract
In an autoimmune environment, B cells spontaneously participate and are selected in germinal centers (GC). In previous work, we investigated epitope spreading in a mixed bone marrow chimera model that combines B cells from lupus-like mice with those from wild type (WT) B6 mice (Degn et al Cell 2017). In the current study, we identify novel biomarkers of autoimmune B cells at the population and single cell level.
Cohorts of autoimmune and WT immunized chimeric mice were prepared. To track WT-derived B cells, we used the tamoxifen inducible AID-CreERT2-EYFP mice. WT chimeras were immunized and boosted. Splenic B cells were sorted based EYFP+ signal and processed for droplet-based gene expression and BCR repertoire single cell RNAseq. To detect specific traits of autoreactive B cells relative to immunized, unsupervised cluster analysis, differential gene expression and pseudotime analysis were used.
Results show higher antibody repertoire diversity in the autoimmune relative to immune B cells, but striking similarity for transcriptomic usage. However, distinct gene expression was identified for the autoreactive B cells such as CXCL10 usage by GC B cells and SLPI expression by autoreactive plasma cells. Importantly, we were able to identify DN2- and DN4-like memory B cells in both conditions. DN2 cells, presumed to derive directly from naïve B cells, have been recently pointed as a source of pathogenic extrafollicular antibody secreting cells in humans.
Overall, results reveal the variety of characteristics that B cells, in their many states, adopt in the response to self and should open the way to new approaches to control their pathogenicity in autoimmune disease.
Role of Extracellular Vesicles in human uveitis
Authors: Agashe Vrushali Vinay , Lee Jung Wha , Maclean Mary , Nath Pulak Ranjan , Sen H. Nida 
Published: February 2023

Abstract
Non-infectious uveitis (NIU) is a significant cause of blindness. It is an immune mediated disease however the exact pathogenesis remains elusive. Considerable evidence exists on the immunomodulatory effects of Extracellular Vesicles (EVs) in various diseases; but little is known on the role of EVs in uveitis patients. We have previously shown that cytokine stimulated ARPE19 cells, release EVs; these EVs are capable of inhibiting T cell proliferation, inducing proinflammatory cytokine production from monocytes and monocyte cell death, in PBMCs from uveitis patients. The purpose of this study was to isolate EVs from uveitis patients and to determine their immune-modulatory potential in-vitro. EVs were isolated from the serum of NIU patients using Size Exclusion Chromatography (SEC) and set up in culture with PBMCs or ARPE19 cells; after stimulation cells were stained to determine activation and cytokine production. A higher number of EVs were isolated from patients, as compared to healthy controls; but EVs from both, were of the appropriate size, had a low protein content and were positive for the exosome associated markers CD9, CD63 and CD81. EVs from patients expressed increased levels of CD8, CD29, CD44, and HLA-DR/DP/DQ, indicating their potential to activate cells in-vivo as compared to controls. An upregulation of CD69 and CD25 on T cells and increased IFNg production was observed when the cells were cultured with EVs from patients. Incubating ARPE19 cells with the EVs from patients, increased the expression of MCP1 and IL8 in the cells. Preliminary data indicates that EVs isolated from uveitis patients can induce a robust activation of cells in-vitro. Current experiments are underway to further substantiate these results.
Dissecting the complexity of heart infiltrates in post-infectious myocarditis induced with CVB3 infection by single-cell RNA sequencing analysis
Authors: Lasrado Ninaad , Borcherding Nicholas , Arumugam Rajkumar , Starr Timothy K. , Reddy Jay 
Published: February 2023

Abstract
Viral myocarditis is a predominant cause of heart failure in children and young adolescents that can lead to dilated cardiomyopathy (DCM). Experimentally, Coxsackievirus B3 (CVB) is commonly employed to study the pathogenesis of myocarditis. Post-infectiously, affected mice can develop DCM, and viral RNA may be present with or without inflammation raising a question as to the events culminating in the development of DCM. To dissect this complexity, we sought to characterize heart infiltrates from CVB infected mice by single-cell RNA sequencing and compared the profiles with healthy mice. These analyses revealed 26 subtypes of cells with fibroblasts, T cells and macrophages as the major constituents. Other cell types include endothelial cells (EC), smooth muscle cells, granulocytes, B cells, and innate lymphoid cells. By analyzing the transcriptomes, we noted differential expression of transcription factors (TFs) in various cell types. One such cell type is EC, in which, activating transcription factor 4, interferon regulatory factor 1 (IRF1) and signal transducer and activator of transcription 6 (STAT6) that have a role in apoptosis and inflammation were upregulated. Correspondingly, expression of Myc and nuclear factor erythroid-derived 2-like 2 implicated in EC survival and angiogenesis was downregulated. The data suggests the ECs may play a role in the pathogenesis of CVB infection through a proinflammatory transcriptional program.
Zfp36 family members redundantly protect against T cell-mediated autoinflammation and premature mortality
Authors: Cook Melissa Erin , Bradstreet Tara R , Webber Ashlee , Kim Jongshin , Santeford Andrea , Schwarzkopf Elizabeth A , Stumpo Deborah J , Apte Rajendra S , Blackshear Perry J , Edelson Brian T 
Published: February 2023

Abstract
Cytokine production must be tightly regulated in order to prevent auto-inflammatory diseases. The zinc finger 36 (Zfp36) family of RNA-binding proteins, including Zfp36, Zfp36l1, and Zfp36l2, are known to negatively regulate mRNA stability or translation of many transcripts, including cytokines. Polymorphisms in ZFP36L1 and ZFP36L2 have been identified in GWAS studies of a variety of human autoimmune diseases, necessitating understanding their functions. While there are reports of all three family members regulating T cell cytokine production, delineating the exact functions of these genes is challenging due to spontaneous phenotypes upon global deletion of single genes and potential redundancy in their functions. To overcome this, we generated Cd4-Cre+ Zfp36fl/fl Zfp36l1fl/fl Zfp36l2fl/fl mice. Only upon triple deletion, but not individual or paired deletions, do mice spontaneously develop an inflammatory disease characterized by early mortality and immune cell infiltration into various organs, including the central nervous system, kidneys, and liver. These mice have drastically elevated levels of many cytokines in their sera. RNA-sequencing reveals that deleted T cells are enriched in gene pathways involving inflammation, proliferation, and apoptosis. Our findings demonstrate a novel redundancy of the Zfp36 family members in regulating T cell homeostasis and suppressing cytokine-driven inflammation. We are investigating the mechanisms and mRNA targets that control this phenotype. Understanding the individual and redundant functions of the Zfp36 family members may lead to opportunities to target them for suppression of T cell-driven autoimmunity or for activating anti-tumor and anti-pathogen T cell responses.
An Exon 2-deficient isoform of the Foxp3 gene fails to protect against IgE-mediated cutaneous autoimmunity after ultraviolet light exposure.
Authors: Domeier Phillip P , Chang Sarah , Utz Paul J , Zhou Baohua , Ziegler Steven F 
Published: February 2023

Abstract
FOXP3 is required to maintain immune tolerance, but the functional contributions of FOXP3 isoforms to the maintenance of tolerance is poorly understood. Mice express a single isoform of the Foxp3 gene, but humans encode an additional exon 2-deficient splice variant isoform of FOXP3 with no known function. To determine how exon 2 of FOXP3 contributes to Treg activity, we generated mice that only express the exon 2-deficient FoxP3 variant (called B6.FoxP3ΔExon2). In the steady-state, B6.FoxP3ΔExon2 mice develop mild autoimmunity (splenomegaly, autoantibodies and kidney nephritis) with enlarged germinal centers and elevated titers of skin-reactive IgE. We also observed skin-reactive IgE antibodies in SLE patients with active photosensivity. Therefore, We challenged B6 and B6.FoxP3ΔExon2 mouse skin with ultraviolet B (UVB) light to determine if UVB damage can increase the production of self-reactive IgE. When compared to B6 controls, B6.FoxP3ΔExon2 mice developed greater skin inflammation with increased infiltration of IgE+ plasma cells (B220lowCD138+Blimp1 +) at the site of UVB damage. B6.FoxP3ΔExon2 mice also had higher numbers of IL4-producing T effector cells (IL4+CD4+CD44+Foxp3−) and IgE-producing B cells in draining lymph nodes of UVB-challenged skin as compared to untreated control lymph nodes. Finally, UVB-treated B6.FoxP3ΔExon2 mice also had worsened kidney nephritis and proteinurea, phenocopying multi-systemic flares that occur in SLE patients with photosensitivity. Collectively, we found that exon 2 of the Foxp3 gene is required to control cutaneous inflammation though the regulation of skin-reactive IgE, and IgE-producing B cells could be targeted to limit the severity of SLE-associated photosensitivity.
Lupus-like extrafollicular B cell responses and autoreactivity are hallmarks of severe COVID-19
Authors: Woodruff Matthew C , Ramonell Richard P , Lee F. Eun-Hyung , Sanz Ignacio 
Published: February 2023

Abstract
In late 2019, the emergence of the coronavirus SARS-CoV-2 resulted in a global pandemic, costing over 2 million lives. A hallmark of the disease resulting from SARS-CoV-2 infection, COVID-19, has been it’s heterogeneity in presentation. To understand the immunological underpinnings of this variability in symptoms, we performed a deep characterization of the B cell responses generated in patients within the NIH-defined mild/moderate or severe/critical categories. Somewhat surprisingly, patients with severe disease had higher neutralizing antibody titers than their mild/moderate counterparts, but showed strong evidence of extrafollicular B cell activation by flow cytometry – previously described in humans primarily in autoimmune disease. Further analysis of the B cell repertoire through single cell VDJ sequencing revealed low (or fully absent) levels of somatic hypermutation in the antibody secreting cell compartment, phenocopying the low selective pressures previously associated with extrafollicular responses. Together, these findings suggested the possible relaxation of peripheral tolerance in severe patients. This was confirmed through clinical autoreactivity testing, revealing more than 70% of the most severe COVID-19 patients, despite no history of autoimmunity, test positive for anti-nuclear antibodies or rheumatoid factor. Altogether, these findings provide the immunological context for an emerging consensus in the literature whereby severe COVID-19 is characterized by robust extrafollicular B cell activation, resulting not only in neutralizing antibody production, but also autoreactive B cell targeting with the potential for direct involvement in severe disease pathology.