bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of Janus Kinase pathway corrects IFN-driven inflammation and epithelial dysfunction in Sjogren’s disease
Authors: Gupta Sarthak , Yamada Eiko , Nakamura Hiroyuki , Khavandgar Zohreh , Martin Daniel , Tandon Mayank , Alevizos Ilias , Jang Shyh-Ing , Perez Paola , Dominick Kalie , Pranzatelli Thomas , Baer Alan , Chiorini John , Warner Blake 
Published: August 2023

Abstract

Background/Purpose
A systematic investigation of the role of interferons (IFNs) and Janus kinases (JAK) - signal transducer and activator of transcription (STAT) pathway at the cellular and tissue level in humans is lacking.

Methods
PBMC and minor salivary gland (MSG) biopsies were obtained from healthy volunteers (HVs) and SjD patients that met the 2016 American College of Rheumatology classification criteria on IRB approved protocols.

Results
RNA sequencing (RNAseq) on MSGs found enrichment of JAK-STAT pathway genes in SjD (n=36) compared to HV (n=13). A composite score of 21-IFN stimulated genes (ISGs) was elevated in SjD and positively correlated with clinical variables of SjD (focus scores). Immunofluorescence showed increased expression of JAK3 in infiltrating lymphocytes. Single cell RNAseq (scRNAseq) of MSGs from SjD (n=12) and HVs (n=17) showed upregulated JAK-STAT pathway genes in infiltrating T and B lymphocytes; antigen presenting cells; and endothelial cells. Ex-vivo studies showed baseline elevation of intracellular phosphorylated STAT1 and STAT3 levels in PBMCs in SjD (n=21) vs HVs (n=11). Stimulation with IFNb further increased the pSTAT1 signaling and JAK inhibition with tofacitinib led to normalization of JAK-STAT signaling. HV and SjD-derived primary salivary gland epithelial cells (pSGEC) showed elevated basal CXCL10 in patients with SjD (n=5) vs HV (n=5); and JAK inhibitors (tofacitinib, baricitinib) blocked IFNb driven ISG expression.

Conclusion
Our findings pinpoint cells responsive to JAK inhibition and illustrate in patient tissues that JAK inhibitors may be beneficial in SjD. These data will serve as biological endpoints for clinical trials [NCT04496960].

This research was supported by the Intramural Research Programs of the National Institute of Arthritis and Musculoskeletal and Skin Disease and National Institute of Dental and Craniofacial Research
In vitro generation of disease-specific autoantibodies produced by B cells from patients with NMOSD and MOGAD
Authors: Payumo Rosah May Palermo , Kim Yeseul , Kim Soyeon , Han Sang-Min , Park Kevin , Park Hyewon , Kim Ha-Eun , Kim Su-Hyun , Hyun Jae-Won , Kim Ho Jin 
Published: August 2023

Abstract
The main pathogenic feature of neuromyelitis optica spectrum disorder (NMOSD) and myelin oligodendrocyte glycoprotein associated disease (MOGAD) is the presence of autoantibodies against water channel, aquaporin-4 (AQP4) and myelin oligodendrocyte glycoprotein (MOG), respectively. However, the underlying mechanisms in which the B cells from these patients produce autoantibodies is largely unknown.
Our study aims to establish a human culture system that could characterize the contribution of B cells in the production of these autoantibodies.
PBMCs were obtained from NMOSD (n=30) and MOG (n=10) patients. B cells were differentiated into plasmablasts in vitro with different stimuli resembling the mechanism related with NMOSD and MOGAD development. After 6d culture, cell proliferation and plasmablast frequency were assessed. Quantification of total IgG levels and detection of AQP4-IgG or MOG-IgG were determined on D13.
Our data showed that the presence of R848 (TLR7/8), regardless of sCD40L or cytokines, resulted in B cell proliferation in NMOSD (ave: 78.2% ± 18.5 SEM) and MOGAD patients (ave: 90.4% ± 5.2 SEM). Moreover, in vitro generation of plasmablasts require sCD40L and R848 in NMOSD (ave: 45.6 ± 21.1 SEM) and MOGAD (ave: 54.4% ± 11.5 SEM). Further culture until D13 resulted in IgG secretion by the plasmablasts generated from NMOSD (ave: 18,150.07 ng/mL) and MOGAD (ave: 17,959.25 ng/mL). For all the conditions that generated plasmablasts, autoantibodies were detected to be reactive with either AQP4-transfected cells in NMOSD (10/30 patients) or MOG-transfected cells in MOGAD (6/10 patients).
This in vitro model have important implications for therapy selection and understanding the mechanisms of NMOSD and MOGAD development.
This study was supported by the International Cooperation Education Program of National Cancer Center Korea (NCCRI·NCCI 52210-52211).
Humanized BAFF transgenic mice develop autoimmune manifestations
Authors: Li Zhiying , Liang Juan , Zhang Yuxi , Xu Shuxin , Liu Qinglong , Tong Huan , Ku Cunxiang , Zhao Jing , Xiang Gao 
Published: August 2023

Abstract
The B cell activating factor(BAFF) is a B cell survival factor that supports autoreactive B cells’ survival and prevents their depletion. Excessive BAFF can increase autoreactive B cells, driving autoimmunity. BAFF is commonly overexpressed in Systemic Lupus Erythematosus(SLE) and is strongly involved in the pathogenesis of the disease. Mice with BAFF deficiency results in the lack of mature B cells, whereas mice that overproduce BAFF have high numbers of mature B cells and antibodies, including autoantibodies, and develop an autoimmune disease similar to SLE in humans. BAFF has been shown to be the pivotal target in autoimmune diseases. In this study, we report the generation of two transgenic mouse strains with overexpression of human BAFF (B6-hBAFF, T036791 and T036794), which could be used to study anti-BAFF therapies.
Overexpression of human BAFF led to significantly increased IgG, IgA, and IgM levels since 6 weeks of age compared with wild-type mice, which continued to increase gradually as the transgenic mice aged. These mice secreted a high level of anti-DNA autoantibodies, along with an increased ratio of B cells/CD45+ in the spleen and lymph node compared with wild-type mice, indicating overreactive B cells. Proteinuria started at 6 weeks of age with an increase in protein/creatinine ratio. This was accompanied by the infiltration of inflammatory leukocytes in the kidney at 15 weeks of age, suggesting local kidney inflammation. Taken together, the humanized BAFF transgenic mice autoimmune manifestations with enhanced overreactive B cells, providing a valuable tool supporting preclinical efficacy studies of drugs targeting B cells for the development of autoimmune disease therapies.
Selective inhibition of estrogen receptor alpha restores regulatory T-cell function in autoimmune hepatitis
Authors: Longhi Maria Serena , Graham Jonathon J , Gao Li , Zhang Wei , Zhang Lina , Nastasio Silvia , Kalbasi Ahmadreza , Bonder Alan , Patwardhan Vilas , Robson Simon C 
Published: August 2023

Abstract
Dysfunction in regulatory T-cells (Tregs) is a hallmark of autoimmune hepatitis (AIH), a serious hepatopathy often leading to end-stage liver damage and transplantation, if not treated. Treg impairment in AIH has been linked to defective response to aryl hydrocarbon receptor (AhR) activation. AhR acts as a modulator of toxin and adaptive immunity. We have previously shown that altered response of Tregs to AhR activation derives from heightened levels of estrogen receptor alpha (Erα), an AhR non-canonical binding partner. In this study we investigated whether Erα selective antagonism modulates Treg function and response to AhR in AIH.
Experiments were performed on immunomagnetically purified Tregs, exposed to the Erα-specific antagonist methyl-piperidinopyrazole (MPP), in combination with or without unconjugated bilirubin (UCB), a known AhR endogenous ligand. In healthy controls (HC), UCB boosted Treg ability to suppress T-responder cell proliferation, and this was further increased when UCB was combined with MPP. In AIH, addition of MPP significantly ameliorated Treg ability to suppress, when exposed to UCB. Stimulation of Tregs with UCB and MPP boosted CYP1A1, CD39 and IL-10 levels both in AIH and HC. Analysis of Treg transcriptome indicated that treatment of Tregs with MPP increased the levels of CD73, an ectoenzyme that works in tandem with CD39, to ultimately generate adenosine.
In conclusion, selective inhibition of Erα restores Treg ability to respond to AhR resulting in increased CD39 levels and ameliorated suppressive function in AIH. This could provide a new therapeutic strategy to boost and maintain immunotolerance in AIH.
Supported by grants from NIH (R01 DK124408, R01 DK108894)
ANATOMY OF THE IMMUNE RESPONSE TO FRANCISELLA TULARENSIS LVS INFECTION IN THE LUNGS
Authors: Okoye G Donald , Kumar Amrendra , Scott Algood Holly M , Joyce Sebastian 
Published: August 2023

Abstract
The respiratory mucosa is under constant immune surveillance because of its vulnerability to infectious diseases. Infection of the lungs with the live vaccine strain (LVS) derived from Francisella tularensis (Ft) subspecies holarctica models pulmonary tularaemia-like disease in mice. Current evidence suggest many immune cells and cytokines respond to Ft LVS infection, but how these various immune system components are integrated to mount a protective response remains unclear. Hence, in a transcriptomics study at single cell resolution, we characterized the acute immune landscape in the lungs of C57BL/6 mice at day 0 and day 7 post intranasal inoculation with Ft LVS. Defining features of the immune response include a robust type 1 immune response characterized by the accumulation of inflammatory neutrophils and the expansion of innate-like effector lymphocytes, primarily interferon-γ producing NKT1, NK, and effector CD8+ T cells. Increased accumulation of MAIT17 over MAIT1 cells is another feature, which is in line with previous findings linking MAIT cells and IL-17 to LVS immune response. Surprisingly, a significant proportion of MAIT17 cells either maintained or upregulated type 1 inflammatory markers. Similarly, MAIT1 cells maintained or upregulated Type 3 inflammatory markers. Additionally, a highly active MAIT cell subset has increased Nr4a1 expression, which encodes Nur77, suggesting a T cell receptor-mediated activation. By contrast, NKT1 cells poorly upregulated Nr4a1 expression but induced Il18r1 expression suggesting, a cytokine-mediated activation. Consequently, we predict that unconventional NKT & MAIT cells integrate innate cues to control tularaemia-like disease caused by Ft LVS infection in the mouse.
VUSM MSTP NIGMS of the National Institutes of Health T32GM007347 (GDO); IO1 BX001444, BX001610, BX00xxx, IK6 BX004595 & RO1 AI137082; IBX000915A (HMA)
Cell-specific P2RX7 promotes the longevity of lung tissue-resident helper CD4 T cells in response to influenza infection
Authors: Santiago-Carvalho Igor , de Gois Macedo Bruna , Vardam-Kaur Trupti , van Dijk Sarah , Salgado Caio Loureiro , da Silva Henrique Borges 
Published: August 2023

Abstract
Development of universal influenza (flu) vaccines requires better knowledge of lung-resident immune responses. That includes flu-specific CD4 T cells, which establish long-term lung residency and induce a diversified local protective response to subsequent infection. Tissue-resident helper (Trh) cells are a subset of lung-resident CD4 T cells that share phenotypic and functional features with follicular helper cells (Tfh). Trh cells serve to amplify lung-resident immune responses, and express high levels of the extracellular ATP (eATP) sensor P2RX7. Considering that eATP levels in the lung increase in response to flu infection, we used T cell-specific P2RX7-knockout (KO) mice to test the role of P2RX7 for CD4 T cell responses to flu. During acute phase of infection, we found that P2RX7 promotes the accumulation of flu-specific CD4 T cells inside the lung tissue. Moreover, P2RX7 promoted this establishment via induction of CXCR3 upregulation. Despite no specific defects in the Trh pool at the acute phase, T cell-specific P2RX7-KO led to a progressive decay of Trh cells. Consequently, lungs of T cell-P2RX7-KO mice had reduced lung B cell responses. Our single-cell RNAseq of lung flu-specific CD4 T cells showed reduced expression of cell survival and mitochondrial genes in P2RX7-KO Trh cells. Our findings suggest that lung eATP sensing by CD4 T cells induces lung-resident CD4 T cell accumulation in response to flu, impacting the survival of specific lung CD4 T cell subsets.
National Institute of Allergy and Infectious Diseases (NIAID) (H.BdS.: R00 AI139381, R01 AI170649).
Upper respiratory tract resident memory CD8 T cells limit viral transmission and provide durable protection
Authors: Michalets Sarah , Uddbäck Ida , Williams M Elliott , Mattingly Cameron , Kost Kirsten N , Lawrence Laurel A , Lowen Anice C , Scharer Christopher D , Kohlmeier Jacob E 
Published: August 2023

Abstract
An effective vaccine against respiratory viruses should not only limit disease in infected individuals, but also restrain viral dissemination among the population. Furthermore, directing vaccine responses to conserved viral epitopes can offer cross-strain protection and potentially limit viral escape from pre-existing immunity. Although studies have demonstrated that intranasal vaccination can generate populations of resident memory CD8 T cells (TRM) throughout the respiratory tract that can rapidly clear a secondary infection and limit immunopathology, the ability of CD8 TRM to limit natural respiratory virus transmission has not yet been evaluated. To address this gap in knowledge, we developed a model of parainfluenza virus transmission whereby a bioluminescent Sendai virus can be used in conjunction with in vivo imaging to track transmission dynamics longitudinally across murine cohorts. Recombinant influenza and adenoviral vectors expressing the immunodominant Sendai virus nucleoprotein CD8 T cell epitope enable the establishment of Sendai virus-specific CD8 TRM throughout the respiratory tract without inducing Sendai-specific antibodies. Using these models, we demonstrate that virus-specific CD8 TRM can inhibit the establishment of infection in immunized hosts and prevent respiratory virus transmission. Additionally, we show that protection against respiratory virus transmission is mediated through TRM localized in the upper respiratory tract and is durable for at least six months. These results indicate that vaccines which induce localized T cell immunity at the initial site of respiratory virus infection can contribute to the generation of herd immunity and limit viral spread in the population.
This project is supported by NIH grants R35 HL150803 and U01 HL139483.
CD11b+Ly6g+CD69+CD74+ cells produce interferon-α in Mycobacterium tuberculosis infected alcohol diet-fed mice
Authors: Mukherjee Tanmoy , Tripathi Ashutosh , Radhakrishnan Rajesh Kumar , Vankayalapati Abhinav , Vankayalapati Ramakrishna , Tripathi Deepak 
Published: August 2023

Abstract
Chronic alcohol consumption is associated with significant mortality and morbidity in people affected by Mycobacterium tuberculosis (Mtb) infection. Our Lab previously reported that alcohol diet increased mortality in young mice infected with Mtb in an Interferon-α (IFN-α) dependent manner. We also found CD11b+Ly6g+ cells are major source for IFN-α. In the current study, using single cell RNA sequencing, we further characterized IFN-α producing CD11b+Ly6g+ cells in the lungs of Mtb infected alcohol-diet fed mice. Our analysis revealed ‘6’ distinct clusters in CD11b+Ly6g+ cell population based on gene signatures. We identified cluster ‘6’, through gene-set scoring and expression of interferon related genes such as IRF1, IRF5 to be responsible for type I interferon production. Pseudotime analysis along with KEGG enrichment, revealed transcriptome of cluster ‘6’ to be in a separate trajectory compared to the core of the cluster, suggesting a differentiated phenotype. Similarly, we identified CD69 and CD74 as a surface marker for this unique cell population which was subsequently verified with t-sne analysis using flow cytometry. We are currently characterizing the cellular mechanisms regulating the expansion of these cells in Mtb infected alcohol-diet fed mice to be able to target them for effective early Interferon-α neutralization.
Intracellular pH controls death and proliferation of lymphocytes in immune response and tumor cells
Authors: Zeng Wei-ping , Yang Shuangshuang , Zhou Baohua 
Published: August 2023

Abstract
Cancer and highly proliferating normal cells have high demand for energy but paradoxically switch energy production from the highly efficient mitochondrial respiration to the low efficient cytosolic glycolysis. This phenomenon is known as the Warburg effect, but the reason for which is not fully understood. This study found that low intracellular pH (pHi) induced apoptosis, whereas high pHi enhanced proliferation, of normal lymphocytes. In tumor cells, low pHi also induced apoptosis whereas high pHi maintained high rates of proliferation. In the draining mediastinal lymph nodes (MLNs) of mice sensitized and challenged with ovalbumin, lymphocytes had dramatically higher pHi and became more heterogeneous than lymphocytes in the non-draining lymph nodes. The pHi of the lymphocytes was inversely related to mitochondrial energetic activities, and the rise of the lymphocyte pHi in the MLNs was accompanied by the reduction of mitochondrial energetic activities. Furthermore, high lymphocyte proliferation in the MLNs powered by strong mitochondrial energy led to low pHi thereby predestined the lymphocytes to death by apoptosis. These findings support a proposed explanation for the Warburg effect as a substitution for mitochondrial respiration that allows the highly proliferating cells to avoid low pHi-induced apoptosis.
None
Sphingosine 1-phosphate / Acetyl CoA Carboxylase1 Axis regulates the Differentiation and Function of Tumor-associated Myeloid-Derived Suppressor Cells
Authors: Chakraborty Paramita , Choi Seungho , Kassir Mohamed Faisal , Oberholtzer Nathaniel , Hedley Zachariah T , Beeson Gyda C , Chatterjee Shilpak , Ogretmen Besim , Mehrotra Shikhar 
Published: August 2023

Abstract
The presence of myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment (TME) poses a serious obstacle to cancer treatment. The bioactive lysophospholipid sphingosine 1-phosphate (S1P), which is produced by sphingosine kinases (SphK1 or SphK2), regulates tumor and T cell survival and activity. Thus, it has been targeted to improve the outcome of anti-cancer therapeutics. However, S1P has not yet been investigated in terms of how it regulates myeloid cell activity. The present study demonstrates that deletion of SphK2-mediated S1P in MDSCs favorably modifies the tumor microenvironment and renders infiltrating myeloid cells with antigen presenting phenotype, leading to increased ability to control tumor growth. Furthermore, in preclinical prostate and melanoma cancer models, the combination of PD1 antibody and SphK2 pharmacological inhibitor (ABC294640) dramatically reduced tumor growth and increased the survival of tumor bearing mice. Mechanistically, the S1P pulldown and subsequent MS-based proteomics analysis revealed that S1P binds to various proteins involved in metabolic and ribosomal biogenesis pathways. Notably, both the isoforms of Acetyl CoA Carboxylase, the rate-limiting fatty acid metabolic enzyme, binds to S1P. Furthermore, investigations using deletion, overexpression, and site-directed mutagenesis demonstrated that ACC1 activity is necessary to preserve the immunogenic capacity of SphK2 KO MDSCs. This study highlights the clinical value of manipulating MDSC suppressive activity by targeting the S1P/ACC1 axis to boost T cell-mediated tumor control and highlights the crucial function that S1P plays as an immunoregulatory mediator in MDSCs.
Supported grants from NIH (R01 CA250458, R01 CA236379)
Poly(lactic acid)-based immunomodulatory nanoparticles promote survival in a polymicrobial mouse model of sepsis
Authors: Truong Nhu , Dharmaraj Shruti , Cottingham Andrea , Shaw Jacob , Lasola Jackline Joy , Scotland Brianna L , Pearson Ryan M 
Published: August 2023

Abstract
Sepsis is a life-threatening multifactorial organ dysfunction caused by a dysregulated host response to infection. Despite being recognized as a global health priority, current therapeutic approaches for sepsis have yet to demonstrate significant improvements in patient outcomes. Our group previously demonstrated that cargo-less immunomodulatory nanoparticles (iNPs) could suppress innate immune cell proinflammatory responses and increase survival in an LPS-induced endotoxemia mouse model. In the present work, we examine the impact of poly(lactic acid)-based iNP delivery on survival and comprehensively characterize the host immune response using a clinically relevant and lethal mouse model of polymicrobial sepsis, cecal ligation and puncture (CLP). iNPs, when administered intraperitoneally in combination with broad spectrum antibiotics following CLP surgery, resulted in a significant improvement in disease symptoms and survival compared to CLP + antibiotics only. We also observed that iNP treatment reduced the levels of proinflammatory cytokines in the peritoneal fluid. Analysis of immune cell populations suggested that iNP treatment modulated macrophage, myeloid-derived suppressor cell, and inflammatory monocyte populations both locally and systemically. Our study demonstrates that local administration of iNPs promotes survival through a multimodal immunomodulatory mechanism to improve sepsis outcomes.
This research was supported by startup funds from the University of Maryland School of Pharmacy, National Institute of General Medical Sciences of the National Institutes of Health under Award Number R35GM142752, and the Shock Society Faculty Research Award awarded to Ryan Pearson. Nhu Truong is supported by a PhRMA Foundation Pre-Doctoral Fellowship from Washington, DC. This publication was supported by funds through the National Cancer Institute - Cancer Center Support Grant (CCSG) - P30CA134274, and Maryland Department of Health’s Cigarette Restitution Fund Program
IL-6 suppresses vaccine responses in neonatal mice by enhancing IL-2 activity on T follicular helper cells
Authors: Parvathaneni Swetha , Yang Jiyeon , Lotspeich-Cole Leda , Sakai Jiro , Lee Robert C , Akkoyunlu Mustafa 
Published: August 2023

Abstract
The inability of neonates to develop CD4+CXCR5+PD-1+ T follicular helper (TFH) cells contributes to their weak vaccine responses. In previous studies, we measured diminished IgG responses when IL-6 was co-injected with a pneumococcal conjugate vaccine (PCV) in neonatal mice. This is in sharp contrast to adults, where IL-6 improves vaccine responses by downregulating the expression of IL-2Rβ on TFH cells and protecting them from the inhibitory effect of IL-2. In this study, we found that splenic IL-6 levels rapidly increased in both adult and neonatal mice following immunization, but the increase in neonatal mice was significantly more than that of adult mice. Moreover, immunized neonatal TFH cells expressed significantly more IL-2 as well as its receptors, IL-2Rα and IL-2Rβ, than the adult cells. Remarkably, IL-6 co-injection with PCV vaccine further increased the production of IL-2 and the expression of its receptors by neonatal TFH cells, whereas excess IL-6 had totally opposite effect in immunized adult mice. Underscoring the role of IL-6 in activating the IL-2 mediated suppression of vaccine responses, immunization of IL-6 knock-out neonates led to improved antibody responses accompanied by expanded TFH cells as well as lower levels of IL-2 and IL-2 receptors on TFH cells. Moreover, CpG containing PCV improved TFH response in neonates by suppressing the expression of IL-2 receptors on TFH cells and inhibiting IL-2 activity. These findings unveil age-specific differences in IL-6 mediated vaccine responses and highlight the need to consider age-related immunobiological attributes in designing vaccines.
This study was funded by US FDA, Perinatal Health Center of Excellence (PHCE) grant to Mustafa Akkoyunlu as well as intramural grants from US FDA.