bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Snap Shot of Complement Gene Expression and Presence of Complement Proteins in Synovial Biopsies from Early Rheumatoid Arthritis Patients
Authors: Banda Nirmal K , Deane Kevin D , Seifert Jennifer , Strickland Colin , Bemis Elizabeth , Jordan Kimberly , Goldmann Kationa ,   Accelerating Medicines Partnership (AMP) RA/SLE Network, Morgan B Paul , Lewis Myles J , Pitzalis Costantino , Moreland Larry W , Holers V Michael 
Published: January 2023

Abstract
The etiology of rheumatoid arthritis (RA) is unknown. Previous studies of mouse models of RA have strongly implicated the alternative and lectin pathways of the complement system in disease pathogenesis. Here we explored the Pathobiology of Early Arthritis Cohort (PEAC) tissue RNA sequencing (RNA-seq) database and identify correlations among complement gene expression, Fc receptor expression and clinical severity, measured as disease activity score 28 - erythrocyte sendimentation rate (DAS28-ESR), in both blood and synovium. We also evaluated the biodistribution of complement activation pathway proteins and inhibitors using Multispectral ImmunoHistoChemical (MIHC) staining. Ultrasound guided synovial biopsies (n = 23), obtained from Accelerating Medicines Partnership (AMP) studies, were subjected to MIHC for various complement proteins. Our analyses revealed that in the synovium, but not in blood, significant positive correlations existed between complement gene expression and DAS28-ESR for C2, CFB, FCN1, C3AR1, C5AR1, and CR1. Surprisingly, levels of MASP1, Colec12, C5 and C6 RNA inversely correlated with baseline DAS28-ESR. After 6 months therapy, baseline CFHR4 positively correlated with delta DAS28-ESR. In the synovium, there were also significant positive correlations between DAS28-ESR and FcγR1A, FcγR1B, FcγR2A and FcγR3A. In early RA (ERA) synovium, a significantly (p &lt; 0.05) higher levels of cells expressed CFH compared with CFB and CFHR4. We also found regional imbalance between C3 and CFH in ERA synovial biopsies. ERA synovial biopsies implicate the complement system in early disease and reveal intriguing differences among factors in clinical relevance, outcome and localized tissue dysregulation.
Supported by R01AR51749-16
Metabolic Dysfunction Governs Regulatory T Cell Inflammatory Response during Inflammatory Bowel Disease
Authors: Bamidele Adebowale , Sagstetter Mary , Hirsova Petra , Ramos Guilherme Piovezani , Westphal Maria , Faubion William A 
Published: January 2023

Abstract

Background
Significant proportion of inflammatory bowel disease (IBD) patients respond inconsistently to therapies, underscoring disease complexity. Interleukin 21 (IL21) is highly expressed during IBD. In addition to T helper (Th) cells, inflammatory regulatory T cells (Tregs) have been linked to refractory IBD. Thus, we explored the metabolic role of IL21 in inducing Treg dysfunction and IBD.

Methods
Human Tregs and Th cells were subjected to transcriptional profiling and metabolic phenotyping. Colitis was induced in Rag1−/− mice by naïve CD4 T cell adoptive transfer.

Results
IL21 stimulation of human Tregs induced glycolysis and OXPHOS. In agreement, IL21 enhanced the expression of genes associated with glycolysis, anabolism, and OXPHOS, leading to inflammatory cytokine production. Mechanistically, we found disruption to mitochondrial integrity with concomitant activation of glycogen synthase kinase 3 (GSK3) β, a kinase known to prevent mitochondrial pyruvate metabolism. IL21-induced GSK3β activation was accompanied by pyruvate and lactate buildup. Notably, GSK3 inhibition or supplementation with membrane-permeable methyl pyruvate abrogated metabolic wiring of and inflammatory responses by IL21-stimulated Tregs and effector Th cells. These results suggest that impaired mitochondrial pyruvate metabolism is a feature of inflammatory CD4 T cells. Lastly, GSK3 inhibition prevented pathogenic CD4 T cell-induced colitis in mice, as evidenced by reduced Mouse Colon Histology Index and serum inflammatory cytokines.

Conclusions
IL21 engages Tregs in a hypermetabolic state that augments inflammatory cytokine production. Therefore, desensitizing CD4 T cells to IL21 may also augment Treg function during IBD.
Supported by NIDDK award K01DK124358, the Center for Cell Signaling in Gastroenterology (P30DK084567), and the Mayo Clinic Center for Biomedical Discovery.
Lung memory B cells promote local IgE production in the respiratory tract
Authors: Nelson Alexander J , Wu Yee Ling 
Published: January 2023

Abstract
Allergen-specific IgE mediates allergic asthma. IgE responses including class-switch recombination (CSR) to IgE are tightly regulated. However, the regulation of local CSR to IgE at mucosal sites is poorly understood. This study aims to elucidate cellular sources that promote IgE CSR and mechanisms that regulate IgE production in the respiratory tract.
To model allergic airway inflammation, we repeatedly administered intranasal ragweed pollen and OVA to mice. We profiled B cell responses to OVA in the lung, the lung-draining mediastinal lymph nodes (medLN), spleen and bone marrow. We used fluorescent reporter mice to track IgE CSR and identify B cell subsets that promote IgE CSR by flow cytometry and microscopy.
We found that intranasal allergen exposure elicited robust local B cell responses with the formation of OVA-specific B cells in the medLN and OVA-specific IgE in the airway. OVA-specific germinal center B cells and memory B cells (MBCs) accumulated in the lungs. Using fluorescent reporter mice, we showed that IgG1+ MBCs are the predominant IgE-switching cells. Importantly, the number of IgE-switching MBCs are the highest in the lungs, and IgG1+ MBCs exhibit an increased frequency of IgE CSR in the lungs compared to lymphoid tissues. These suggest that the lung environment favors IgE production. RNA-seq analyses comparing lung and circulating MBCs revealed elevated expression of Tnfsf4 (OX40L) and Icosl on lung MBCs, suggesting a higher propensity for lung MBCs to engage T cells.
Together, these data demonstrate that the allergic lung is a major site of IgE CSR with IgG1+ MBCs promoting local IgE CSR. Our ongoing work focuses on delineating the B cell-intrinsic and extrinsic factors that promote IgE production in the respiratory mucosa.
Supported by grants from NIH (1F31 HL156459, 5T32 AI007508, 1R21 AI159456)
VEGFR3-driven pulmonary lymphangiogenesis exacerbates induction of bronchus-associated lymphoid tissue in allergic airway disease
Authors: Gomez Medellin Jorge E , Hollinger Maile Kananiokala , Rosenberg Jillian , Blaine Kelly , Kurtanich Trevin , Ankenbruck Nick , Hrusch Cara Lynn , Sperling Anne I , Swartz Melody A 
Published: January 2023

Abstract
Asthmatic lung samples present with both a higher density of pulmonary lymphatic vessels and a higher incidence of bronchus associated lymphoid tissue (BALT). Here, we asked whether lymphangiogenesis, stimulated by the VEGF-C/VEGFR-3 signaling axis in lymphatic endothelial cells (LECs), plays a role in promoting BALT in mouse models of allergy. First, we determined that chronic intratracheal instillation of house dust mite (HDM), a clinically relevant allergen, recapitulates both lymphangiogenesis and BALT induction. Intratracheal stimulation of VEGFR-3 in LECs exacerbated BALT, while blocking VEGFR-3 signaling reduced BALT. Furthermore, in transgenic mice with an expanded pulmonary lymphatic network (induced prior to allergen challenge), we found an exacerbated BALT response upon chronic HDM inhalation. Recent studies have determined that LEC-derived CXCL13 plays an important role in secondary lymphoid structure organogenesis, and thus we pondered whether LEC-derived CXCL13 could play a role in the development of tertiary lymphoid structures. Indeed, we observed an increase in lung infiltration by CXCR5+ cells when we used VEGF-C to modulate the chronic allergic response. Finally, we found that the VEGF-C exacerbated BALT phenomenon was indeed CXCL13 dependent. Altogether, these results suggest a causative role for pulmonary lymphatics in mediating BALT induction in chronic allergic airway inflammation.
Choline acetyltransferase attenuates inflammation in a murine model of DSS colitis and sepsis
Authors: Tsaava Tea , Tynan Aisling , Hepler Tyler D , Gabalski Arielle H , Hide Daniel M , Li Jian Hua , Chang Eric H , Tracey Kevin J , Chavan Sangeeta S 
Published: January 2023

Abstract
Excessive immune cell activation and cytokine release leading to inflammatory conditions are associated with bidirectional immune system-brain communication and other physiological responses. The vagus nerve conveys sensory information to the brain and brain derived immunoregulatory signals suppressing peripheral cytokine levels and inflammation. Acetylcholine (A mediated cholinergic signaling has been implicated in this regulation. However, the possibility of controlling inflammation by peripheral administration of choline acetyl transferase (ChAT), enzyme that catalyzes biosynthesis of acetylcholine, is unexplored. We studied the administration of ChAT in endotoxemia, sepsis and DSS colitis models. Intraperitoneal administration of ChAT significantly (p=0.002) suppresses serum levels of TNF (ChAT 1062.0pg/mL ± 113.0 vs vehicle 1711.0pg/mL ± 210.2) during murine endotoxemia. In a murine CLP sepsis model, administration of pegylated ChAT (PEG-ChAT) significantly improves survival (p=0.015), with a 55.6% survival in PEG-ChAT treated mice compared to a 22.2% survival in vehicle mice. In a preclinical model of inflammatory bowel disease, ChAT administration significantly improves body weight gain (d14 p=0.01 ChAT mice -5.1%±1.1 vs vehicle mice -15.5%±2.9), disease score (d11 p=0.006 ChAT mice 0.17DAI±0.05, vs vehicle mice 0.66DAI±0.13), and colon length (p=0.006 ChAT mice 7.15cm±0.2 vs vehicle mice 6.2cm±0.2) compared to vehicle. These results indicate that administration of ChAT inhibits TNF levels in acute endotoxemia and attenuates disease severity in murine models of sepsis and DSS-induced colitis, suggesting that further study of ChAT as an experimental anti-inflammatory therapeutic is warranted.
Supported by grant from NIH to KJT and SSC.
PD-L1 plays a B cell-intrinsic role in suppressing the antibody response and anti-tumor immunity.
Authors: Cervantes Christian , Khan Mustafa , Fernandez Maria , Liu Zexuan , Hakeem Zainab , Viswanadhapalli Suryavathi , Gupta Harshita B , Curiel Tyler J , Yan Hui , Vadlamudi Ratna , Xu Zhenming 
Published: January 2023

Abstract
The discovery that PD-L1 inhibits PD-1 on CD8+ T cells has led to new mechanistic insights into the regulation of host immune responses. The contribution of PD-L1 expressed by different immune cell types remains poorly understood, partly due to the lack of cell type-specific Cd274 (encoding PD-L1) knockout mice. Here, to address the role of B cell-expressed PD-L1 in antibody response and tumor development, we have generated Aicdacre Cd274fl/fl mice, in which PD-L1 was ablated only in activated B cells. As compared to their wild-type littermates, Aicdacre Cd274fl/fl mice showed normal development and maintenance of B cells, T cells, DCs and macrophages, while mounting a stronger T-dependent high affinity NP-binding IgM, IgG1 and IgG2a titers upon immunization with NP-CGG plus alum. Such phenotype was copied by C57 mice treated with a blocking αPD-L1 antibody. Tumor growth from engrafted E0771 breast cancer cells in Aicdacre Cd274fl/fl mice was significantly delayed, in association with increased CD8+ T cell infiltration and activities. Moreover, B cells isolated from Aicdacre Cd274fl/fl mice or those treated with aPD-L1 displayed higher levels of proliferation and differentiation to plasma cells upon in vitro stimulation with the T-dependent B-cell stimulus CD154. This along with the altered gene expression in Aicdacre Cd274fl/fl mice and αPD-L1-treated cells suggests a B cell-intrinsic role of PD-L1 in regulating B cell differentiation. Furthermore, PD-L1 expressed on B cell surface was tightly regulated during differentiation and internalized upon αPD-L1 treatment, suggesting a possible new mechanism on how PD-L1+ B cells modulate host immune responses.
Supported by grants from NIH: The Ruth L. Kirschstein NRSA Institutional Research Training Grant (to MK), NIH T32 Graduate Research in Immunology Program (to CC), R01 AI153506 and R21 AI135599 (to ZX). Mays Cancer Center Dew Foundation Pilot Grant (to ZX) and DoD BCRP BC170448 (to ZX)
Activation of C-type lectin receptor DC-SIGN increases human B cell survival
Authors: Arias Ivonne Nannette , Valentín-Acevedo Anibal José 
Published: January 2023

Abstract
DC-SIGN is a C-type lectin that plays an important immune role as a pattern recognition receptor (PRR). It is mainly expressed in dendritic cells, macrophages, and activated B cells where it can bind several pathogens including HIV and SARS-CoV-2. Interestingly, some of these pathogens use DC-SIGN to facilitate infection of target cells and support their survival within the host. While the role of DC-SIGN as a PRR is well established, little is known about the cellular outcomes induced upon ligand binding, particularly in B cells. Here, we demonstrate that when we activate DC-SIGN with a mannose ligand (mannan) on a lymphoblastic human B cell line with constitutive DC-SIGN expression (Ramos-DC), the cell’s survival increases. Moreover, analysis of cell death by flow cytometry showed that anti-IgM induced death was partially prevented by pre-treating cells with mannan. While mannan can bind to other lectins (e.g., Mannose Receptor, Dectin-1), preliminary immunophenotype analysis of these receptors confirmed their absence from Ramos-DC cells supporting the specificity of our observation. Finally, treating cells with an anti-DC-SIGN specific antibody before inducing death also prevented cells from dying, suggesting that the survival response observed is indeed mediated by DC-SIGN ligation. Our data suggest that DC-SIGN activation in human B cells induces a signaling pathway that results in cell survival even in the presence of death-inducing stimulation. Our future goal is to characterize the signaling events leading to this response in B cells. Our results contribute to the understanding of the role of DC-SIGN in immune function and how this knowledge could lead to the development of novel anti-viral therapies.
The project described was supported by Award Number R25GM110513 from the National Institute of General Medical Sciences. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the National Institutes of Health.
MICRORNA-10A NEGATIVELY REGULATES INTESTINAL IGA RESPONSE THROUGH SUPPRESSING REGULATORY T CELLS CONVERSION TO T FOLLICULAR REGULATOR CELLS
Authors: Yu Tianming , Yang Wenjing , Yao Suxia , Cong Yingzi 
Published: January 2023

Abstract
Intestinal IgA response is a key immune regulation between host and gut microbiota. Regulatory T cells (Tregs) have been recently shown to promote IgA responses through conversion to T follicular regulator cells (TFRs). However, how Treg converts to TFR to promote intestinal IgA response remain unclear. Tregs express microRNA (miR)-10a at high levels, however, whether it regulates TFR generation and function is unknown. We found that Treg cell-specific miR-10a deficient Foxp3YFP-cre miR-10af/f (Foxp3ΔmiR-10a) mice displayed higher levels of gut IgA compared to wild type (WT) Foxp3YFP-cre miR-10af/+ mice. TFRs in Peyer’s patches were also significantly increased while there were no differences in germinal center B cells, Tregs, and TFH cells. When orally immunized with cholera toxin (CT), Foxp3ΔmiR-10a mice showed more CT-specific fecal IgA and TFRs. miR-10a deficient Tregs induced more IgA compared to WT Tregs when co-cultured with naïve B cells in vitro. When transferred to TCRβxδ−/− mice, miR-10a deficient Tregs induced more gut IgA and higher levels of TFRs compared to WT Tregs in vivo. However, gut microbiota composition in Foxp3ΔmiR-10a mice was not altered by 16S rRNA gene sequencing. RNA-sequencing analysis showed that miR-10a was enriched in the oxidative phosphorylation (OXPHOS) process in Tregs. Consistently, miR-10a deficient Tregs displayed higher OXPHOS metabolic levels by seahorse analysis. Pretreatment of Tregs with oligomycin, which inhibits OXPHOS, inhibited Treg induction of IgA production in vitro, indicating miR-10a suppressed Tregs induction of IgA ability by inhibiting OXPHOS. Thus, miR-10a negatively regulates TFRs conversion through inhibition of OXPHOS to control intestinal IgA response.
Burns are more than skin deep: Heightened neuroinflammation after cutaneous injury in aged mice
Authors: Walrath Travis Michael , Najarro Kevin M , McMahan Rachel H , Idrovo Juan-Pablo , Quillinan Nidia , Kovacs Elizabeth J 
Published: January 2023

Abstract
By 2050, 1 in 6 people will be 65 or older. As the aging population grows so too does the importance of understanding how injuries in this age group lead to a greater incidence of neurological complications and mortality. Neurological complications may be driven by elevated pro-inflammatory cytokines and astrocyte activation within the hippocampus (HPC), which reduces levels of phosphorylated cyclic-AMP response binding element (pCREB), leading to cognitive decline and memory impairment. To examine the effects of injury on neuroinflammation in advanced age, young and aged mice were anesthetized and subjected to a 15% total body surface area scald or sham injury. As an index of neuroinflammation, we observed an 85-fold (p&lt;0.05) increase in ccl2 and a 35-fold (p=0.05) elevation in tnfa 24 hours post injury in the brains of aged mice compared to young groups and aged sham mice. We also saw a 13-fold (p&lt;0.05) increase in ccl2 within isolated hippocampus of aged injured mice, when compared to aged shams. Interestingly, when investigating specific HPC regions, immunofluorescence staining showed a 20% rise (p&lt;0.05) in glial fibrillary acidic protein (GFAP) positive astrocytes within the cornu ammonis 3 (CA3) of aged injured mice when compared to sham groups. GFAP+ astrocytes are associated with neurotoxicity and cognitive decline. Finally, there was a 20% decrease (p&lt;0.05) in pCREB staining by immunohistochemistry in the dentate gyrus of aged mice compared to young regardless of injury. These data suggest that remote injury in aged, but not young mice, causes neuroinflammation and astrocyte activation, which paired with an age related reduction in pCREB, could contribute to cognitive decline.
Supported in part by NIH R01 AG018859 (EJK), R35 GM131831 (EJK), T32 AG000279 (KLM), and K08 134185 (JPI)
Clonal and Diversity Differences of Extravascular B cells in the Influenza-Experienced Lung
Authors: Breen Michael P , Feng Feng , Aihara Fumiaki , Etesami Neelou , Fearns Rachel , Mizgerd Joseph , Kepler Thomas B 
Published: January 2023

Abstract
Memory B cells are a vital arm of the antiviral response. Recent literature has characterized a population of murine lung resident memory B cells after influenza A virus (IAV) infection capable of in situ differentiation into anti-influenza antibody secreting cells. Separately, PDL2+/CD80+ B cells appear to have a “memory-like” phenotype while the double-negative counterparts are “naïve-like” with low mutation frequency and class-switching. Additionally, our group has shown that PDL2+ EBCs impact Streptococcus pneumoniae immunity but their role in anti-viral immunity is understudied.
While extravascular B cells (EBCs) may play a critical immune function, it’s unclear if the PDL2+/CD80+ population is clonally distinct from the PDL2-/CD80- one as diversity can impact immune capabilities. We hypothesize PDL2+/CD80+ EBCs have a signature clonal repertoire influenced by infection history and tissue localization.
We infected BALB/cJ mice twice with related IAVs separated by four weeks. After a six-week rest period, we isolated PDL2-/CD80- and PDL2+/CD80+ EBCs from the lung and spleen. Immunoglobulin sequencing (IgSeq) was performed on IgM and IgG libraries that were analyzed with in-house clonal partitioning software (Cloanalyst).
There was significant increase in the number of EBCs in the lung and the proportion of PDL2+/CD80+ EBCs increased dramatically. Mutation frequency of PDL2+/CD80+ EBCs within the IAV-exposed lung rose, suggesting a role of tissue localization on B cell signature. Furthermore, diversity analysis using Shannon and Simpson estimators suggests differences between tissues and PDL2/CD80 expression. Together, our data highlight a unique clonal niche occupied by PDL2+/CD80+ EBCs.
Supported by grants from NIH (R33 HL-137081, T32 AI-007309)
Lung tissue resident memory CD8 T cells rely on lipid-rich environments for differentiation and display unique metabolic profiles
Authors: Sportiello Michael John , Geber Adam , Poindexter Alexis , Reilly Emma C , Emo Kris Lambert , Topham David J 
Published: January 2023

Abstract
Tissue resident memory CD8 T cells (TRM) protect against infections. In the lungs, TRM offer critical protection against influenza. Skin TRM display a unique metabolic profile compared to other memory cell subsets. They undergo enhanced fatty acid oxidation and oxidative phosphorylation as compared with other cells.
It is unclear if this programming holds true in the lung. We hypothesize that lung CD8 T cells with different integrin profiles will utilize unique metabolic programs. Differential expression and pathway analyses were conducted on RNAseq datasets, demonstrating an increased reliance on fatty acid oxidation in TRM compared to circulating memory CD8 T cells. This was most clear when a novel pathway analysis software ‘fluximplied’ that we created was used.
Metabolic models were constructed to verify these findings, functional assays were conducted, and the levels of oxidative phosphorylation, mitochondrial mass, and neutral lipids were measured in addition to fatty acid uptake, glucose uptake, and fatty acid oxidation Seahorse assays, confirming a lipid-centric metabolism of TRM.
It is unclear if this lipid-centric nature of TRM is a correlate of TRM phenotype or essential to TRM development. We developed an ex vivo model where naive T cells were differentiated with cytokines and metabolites into integrin expressing subsets. Higher amounts and more time with lipid-rich media increased expression of TRM markers. Lastly, in a T cell transfer model, we demonstrate that naive T cells that uptake more fatty acids have more TRM progeny than their low-uptake counterparts after transfer and influenza infection. In summary we demonstrate lung TRM have unique metabolic profiles and that lipids impact TRM differentiation patterns.
This work was funded by a Program Project grant through the National Institutes of Health, National Institute of Allergy and Infectious Diseases P01-AI102851 and through the National Institutes of Health training grant T32GM007356-46 and the National Institutes of Health training grant T32HL066988-20.