bluuebunny/crossref_metadata_2025_split huggingface dataset:
Uptake of extracellular complement C3 by cardiomyocytes intervenes the intrinsic apoptosis.
Authors: Fang Zhou 
Published: January 2023

Abstract
A recycling pathway of complement C3 was discovered as a component of the intracellular complement system. Human immune cells (B and T lymphocytes) and non-immune cells (endothelial cells) can rapidly internalize C3 from the extracellular milieu, which plays an important role in T cell survival and epithelial cell death. We investigated if the uptake of extracellular C3 can take place in the parenchymal cells which shielded by endothelial/epithelia barriers and further studied the pathophysiological role of such internalized C3. Using an in vitro system of human cardiomyocytes, we found that AC-16 cardiomyocytes, which did not produce matured C3, were readily uptake the exogenous C3 once being exposed to it. Such internalized C3 helped these cells to mitigate apoptosis under oxidative stress condition. Cardiomyocytes may be exposed to circulating C3 after the endothelial barrier being compromised, which happened upon the reperfusion of ischemic myocardium. We further investigated in the effect of internalized C3 in an animal model of myocardial ischemia/reperfusion (IR). Our results showed that C3−/− mice had significant increase of apoptosis compared with WT mice. Furthermore, C3-bound immunocomplex from WT mice undergone IR contained Cyt c, a key apoptotic factor in the intrinsic apoptosis pathway. Biochemically, C3 purified from blood can inhibit the Cyt c mediated cell death in a cell free apoptosis system. Thus, the C3-uptaking mechanism by the parenchymal cells may serve as an innate immune surveillant for intracellular homeostasis by recognition of potential intracellular damage-associated molecular pattern molecules (DAMPs).
Intracellularly Delivered Nuclear Localization Sequence Attenuates The Severity Of COVID-19 By Inhibits Interaction Of Inflammatory Transcription Factors And Importins
Authors: Kim Dongho , Kim Gyunam , Shin Dasom , Yu Sanghyeon , Baek Misuk , Lee Jaewook , Yu Hyemin , Pi Joohyun , Kang Mingu , Kim Jieun , Kim Sujeong , Chung Eunna , Jo Daewoong 
Published: January 2023

Abstract
The outbreak of infectious severe and acute respiratory syndrome (SARS)-CoV-2 which induces coronavirus disease 19 (COVID-19) causes patients to develop acute respiratory distress syndrome (ARDS) and death by triggering uncontrolled immune responses. Dysregulated immune responses impair healthy organs by over-production of pro-inflammatory cytokines, which are called “cytokine storm”. In this study, an improved cell-permeable nuclear import inhibitor (iCP-NI) has been developed by fusing sequence-optimized advanced macromolecule transduction domains (aMTDs) and nuclear localization sequence (NLS) originated from human NF-κB to treat the cytokine storm mediated organ failure and death in COVID-19. Intracellularly delivered NLS inhibits the transport of various inflammation-associated transcription factors (IATFs) such as NF-κB, NFAT, AP-1 and STAT1/3 which results in suppressed expression of pro-inflammatory cytokines. In brief, iCP-NI decreased the lethality of RNA virus infection mimetic pneumonitis animals by protecting the lung structure from inflammation-mediated destruction via reduction of the inflammatory cytokines in the lung. Continuously, iCP-NI significantly attenuates the severity of the SARS-CoV-2 infected syrian hamster model. The inhibition of viral replication is proved with quantitative PCR by reducing the level of RdRp gene (log7.04 vs log5.79). Likewise, the levels of inflammatory cytokines such as TNF-α and IL-17A in the lungs are significantly decreased and thereby, the lung structures are protected from inflammation-mediated damages. As a result, the iCP-NI could be developed as a novel medicine to cure various inflammatory diseases including COVID-19.
Rad52 mediates class-switch DNA recombination to IgD
Authors: Xu Yijiang , Zhou Hang , Post Ginell , Zan Hong , Casali Paolo 
Published: January 2023

Abstract
While the biology of IgD begins to be better understood, the mechanism of expression of this phylogenetically old and highly conserved Ig remains unknown. In B cells, IgD is mainly expressed together with IgM through alternative splicing of primary VHDJH-Cμ-s-m-Cδ-s-m RNAs, but also independently through IgD class switch DNA recombination (CSR) via double-strand DNA breaks (DSBs) and synapse of Sμ with σδ. How such DSBs, however, are resolved is still unknown. Our previous demonstration of a novel role of Rad52 in a Ku70/Ku86-independent “short-range” microhomology-mediated synapsis of intra-Sμ region DSBs led us to hypothesize that this homologous recombination DNA annealing factor is also involved in short-range microhomology-mediated alternative endjoining (A-EJ) recombination of Sμ with σδ. We found that induction of IgD CSR downregulates Zfp318 and promotes Rad52 phosphorylation and recruitment to Sμ and σδ, leading to alternative end-joining (A-EJ)-mediated Sμ-σδ recombination of resected DSB ends with extensive microhomologies, VHDJH-Cδs transcription and sustained IgD secretion. Rad52 ablation in mouse Rad52−/− B cells aborted IgD CSR in vitro and in vivo and dampened the specific IgD antibody response to OVA. Rad52 knockdown in human B cells also abrogated IgD CSR. Finally, Rad52 phosphorylation was associated with high levels of IgD CSR and anti-nuclear IgD autoantibodies in lupus-prone mice and patients with lupus. Our findings thus show that Rad52 mediates IgD CSR through microhomology-mediated A-EJ in concert with Zfp318 downregulation. This is a previously unrecognized and critical role of Rad52 in mammalian DNA repair that provides a mechanistic underpinning to CSR A-EJ.
Supported by NIH grants AI 079705, AI 105813, AAI 167416 and LRA grant 641363 to PC.
The mutation PTPN6Ala455Thr causing pulmonary emphysema affects B lymphocyte populations and causes spontaneous pulmonary tertiary lymphoid tissue formation
Authors: Pineault Marie , Bouffard Gabrielle , Milad Nadia , Tremblay Félix , Lechasseur Ariane , Routhier Joanie , Aubin Sophie , Beaulieu Marie-Josée , Bossé Yohan , Maltais François , Morissette Mathieu C 
Published: January 2023

Abstract

Background
A new mutation responsible for causing severe and early pulmonary emphysema was recently discovered in a French-Canadian family. The mutation is located in the PTPN6 gene (PTPN6Ala455Thr) leading to a reduction in the activity of SHP-1, a phosphatase that regulates numerous pathways including B cell differentiation and activation. Here, we aimed to characterize B cell abnormalities in both humans and mice carrying this mutation.

Methods
Circulating B cell populations in humans carrying the PTPN6Ala455Thr mutation were analyzed by flow cytometry and immunoglobulin levels were measured in serum. In mice carrying the same mutation (Ptpn6Ala457Thr) aged up to 12 months, B cell populations were analyzed by flow cytometry and lung tissue histology performed. Mice were immunized with the T cell-independent antigen NP-Ficoll and plasma was collected to assess NP-specific antibodies.

Results
Human carriers of the PTPN6Ala455Thr mutation had reduced IgG1 and IgG4, and increased IgG3 levels as well as high circulating immature/mature B cell ratio compared to controls. Aging Ptpn6Ala457Thr mice developed spontaneous pulmonary tertiary lymphoid tissues and exhibited higher B-1 cells and lower B-2 cells proportions in the lung and spleen. Ptpn6Ala457Thr mice had lower levels of NP-specific IgG1 antibodies compared to the wild-type group, but normal IgG3 in response to NP-Ficoll immunization.

Conclusion
SHP-1 appears to be important to B cell development and functions, as the PTPN6Ala455Thr mutation seems to cause a form of subclinical immunodeficiency that affects B cell populations and immunoglobulin levels. The link between B cell abnormalities and development of pulmonary emphysema requires further investigation.
Supported by grants from Canadian Institutes of Health and Research and Fonds de Recherche du Québec- Santé
Effect of post-exercise cupping therapy on the inflammatory markers in taekwondo athletes
Authors: Fang Shih-Hua , Yeh Yu-An 
Published: January 2023

Abstract
Cupping therapy has been claimed to drain toxins, loosen adhesions, and bring blood flow to skin and muscles. Moreover, cupping therapy is believed to modulate the immune system. We investigated the acute effects of post-exercise cupping on the fatigue, mucosal and blood immune responses of atheletes. Fourteen elite taekwondo athletes were recruited, and randomly allocated to two groups including placebo group (PLA) who lied down and rested for 15 min and the treatment group (CUP) who received cupping treatment for 10 min and rested for 5 min. A rated perceived exertion (RPE) was used to evaluate how hard the athletes felt at the time before training (Pre), after training (Post) and 30 min after cupping treatment (Post30). Furthermore, the concentrations of tumor necrosis factor alpha (TNF-a), white blood cells (WBC) and neutrophils, neutrophil–lymphocyte ratio (NLR), and platelet–lymphocyte ratio (PLR) in blood were measured. In addition, the salivary immunoglobulin A (SIgA) and a-amylase activity were also analyzed. The CUP group showed significantly lower RPE than PLA group at Post30. The concentrations of WBC, neutrophil, NLR and PLR were markedly increased after training in both groups, but PLR was significantly decreased at Post30 in CUP group. In addition, the concentration of TNF-a was also decreased significantly at Post30. Furthermore, the activity of a-amylase of CUP group was significantly higher than PLA group at Post30. After high intensity of training, cupping treatment decreased the concentrations of PLR and TNF-a, but increased the salivary a-amylase activity. This study suggested that cupping therapy improved the mucosal immune function and decreased inflammation.
Supported by MOST 106-2410-H-028-002 & MOST109-2410-H-028-009-MY3
Treatment with an orally delivered non-replicating, non-colonizing strain of Veillonella parvula resolves systemic inflammation in murine models of disease
Authors: Ramani Kritika , Cormack Taylor , Cartwright Adam , Alami Aula , Argueta Shannon , Raghunathan Divya , Abdou Mary , Romano Fabian B , Kravitz Valeria , Rommel Tyler , Kiran Esra Uckun , Pradeep Pallavi , Ponichtera Holly , Sizova Maria , Ganguly Tanmoy , Itano Andrea 
Published: January 2023

Abstract

Introduction
The mucosal immune system comprises of a complex network of intestinal epithelial cells and immune cells. We report that an orally delivered microbial strain of Veillonella parvula (EDP1867) that is non-replicating and non-colonizing acts through pattern recognition receptor mediated modulation of small intestinal immune responses to induce potent systemic inflammation resolution.

Methods
EDP1867 is a strain of Veillonella parvula isolated from mucosal tissue of a human donor and rendered non replicating by gamma irradiation. In vitro experiments were performed in primary human immune cells. In vivo efficacy was done using mouse models of delayed-type hypersensitivity (DTH) and psoriasis. Fluorescently labeled EDP1867 was used in biodistribution experiments.

Results
EDP1867 induced IL-10 in primary human immune cells. In a DTH model, orally delivered EDP1867 reduced inflammation and diminished production of pro-inflammatory cytokines ex vivo. Fluorescently labeled EDP1867 transited through the gut without systemic exposure in biodistribution studies to demonstrate gut restriction. EDP1867 activity is dependent on TLR2 and migration of immune cells through the gut. In an imiquimod-induced psoriasis model, EDP1867 reduced skin inflammation and psoriasis score and Th17 cytokines in the skin.

Conclusions
EDP1867 drives broad based resolution of inflammation and can reestablish normal homeostasis. These data demonstrate that EDP1867 can act as a new oral anti-inflammatory drug that is gut restricted and dependent on local immune recognition within the small intestine, to induce systemic anti-inflammatory effects. EDP1867 is currently in clinical development for treatment of inflammatory diseases.
The role of immune checkpoint PD-1 in the submandibular gland
Authors: Borys Samantha , Brossay Laurent 
Published: January 2023

Abstract
Immune checkpoints play an important role in restraining the immune response, maintaining self-tolerance and preventing excessive collateral damage. Immune checkpoint blockade has revolutionized cancer immunotherapy but can lead to immune related adverse events (IrAEs). Patients receiving programmed cell death protein 1 (PD-1) blockade can develop IrAEs including vitiligo, colitis, endocrinopathies, and Sicca Syndrome, an autoimmune disease affecting the salivary and lacrimal glands. After immune cell activation, PD-1 is upregulated, and PD-1 signaling results in reduced proliferation, cytokine production, and cytotoxicity. PD-1 is expressed on infiltrating lymphocytes of the submandibular gland (SMG) including natural killer (NK) and T cells, and recognizes the ligand, PD-L1. Hyporesponsive immune cells in the SMG are thought to retain the integrity of this delicate tissue, while inadvertently contributing to viral latency. We therefore hypothesized that signaling through coinhibitory receptors, including PD-1, may impact these hyporesponsive effector cells. Our preliminary data indicate that genetic loss of PD-1 results in increased CD8+ T cell number and frequency in the SMG, with greater proliferation status. Given the interplay between self-tolerance and chronic viral control, we also examined the role of PD-1 during latent murine cytomegalovirus (MCMV) infection in the SMG. Interestingly, an increased infiltration of CD8+ T cells in the SMG was also observed during MCMV latency period. These studies of PD-1 on SMG immune cells should provide insights for prevention and treatments of IrAEs and chronic viral infections.
This work was supported by the following NIH research Grants: R01 AI46709 (L.B.), R01 AI122217 (L.B.) and a Brown University Presidential Award (S.B.).
Androgens protect ILC2s from interferon-mediated functional suppression during influenza virus infection
Authors: Kovats Susan , Karlik Anna , Turner Sean , Miller Reegan , Ainsua-Enrich Erola , Hatipoglu Ibrahim , Bagavant Harini , Alberola-Ila Jose , Pelikan Richard , Kadel Sapana 
Published: January 2023

Abstract
Biological sex differences in morbidity upon influenza A virus (IAV) infection are linked to stronger interferon-centered immune responses in females, yet the regulatory role of sex hormone receptors in immune cell subsets is incompletely understood. Lung-resident group 2 innate lymphoid cells (ILC2s) express notably high levels of androgen receptors (AR). In IAV infection, ILC2s produce type 2 cytokines and facilitate tissue repair, but they also may be functionally suppressed by type 1 inflammation. Here we report sex differences in the magnitude of lung ILC2 functional suppression at the peak of sublethal IAV infection. Relative to males, ILC2s in female lungs showed attenuated proliferation, decreased propensity for IL-5 and amphiregulin production and reduced expression of GATA3 and IL-33R. Single cell RNA sequencing revealed sex-divergent transcriptomes, with female cells preferentially present in clusters of stressed, functionally suppressed ILC2s and male cells dominating in clusters of canonically activated ILC2s. Equivalent inflammatory cytokine levels and viral load suggested sex differences in ILC2-intrinsic factors. Indeed, naïve female ILC2s showed elevated IFNGR expression and higher phospho-STAT1 levels following IFNG stimulation, and lymphocyte-restricted STAT1 deficiency reversed IAV-induced suppression of female ILC2s. Lymphocyte-restricted AR deficiency or loss of androgens via orchiectomy led to increased IFNGR expression and suppression of male ILC2s. Collectively, these data show that ILC2-intrinsic AR activity regulates IFNGR-STAT1 signaling pathways to preserve canonical ILC2 function in males during IAV infection.
Supported by grants from NIH (HL 119501) and the Presbyterian Health Foundation
Functional tuning of commensal-specific lymphocytes by nociceptive sensory neurons
Authors: Kulalert Warakorn , Wells Alexandria , Enamorado Michel , Link Verena , Kabat Juraj , Kamenyeva Olena , Belkaid Yasmine 
Published: January 2023

Abstract
The somatosensory nervous system surveils external stimuli at barrier tissues, regulating innate immune cells under infection and inflammation. The roles of nociceptive nerves in homeostatic adaptive immunity to the microbiota, however, remain elusive. Here, we identified a novel mechanism for direct neuro-immune interaction between commensal-specific T lymphocytes and skin-innervating nociceptive neurons through secretion of calcitonin gene-related peptide (CGRP). We observed upregulation of the CGRP co-receptor, receptor activity modifying protein 1 (RAMP1), in T lymphocytes induced by topical colonization of the human skin commensal Staphylococcus epidermidis, but not in T cell populations elicited by cutaneous infection or inflammation. Utilizing intravital imaging, we observed that commensal-specific T cells interact intimately and dynamically with skin nociceptive nerves in a CGRP-dependent manner. Importantly, CGRP secreted from nociceptors acts directly on commensal-specific T cells via RAMP1 to regulate their accumulation in the tissues, cytokine production and motility in vivo. Activation or inhibition of nociceptive neurons functionally tune commensal-specific T cells and optimize their responses to ensuing heterologous infections or damaging challenges in the skin. The ability of the somatosensory neurons to participate in the highly specific and potentially long-lasting adaptive immune response to the microbiota underscores the functional versatility and plasticity of commensal-specific T lymphocytes that can be swiftly tuned to diverse arrays of sensory modalities ranging from thermosensation to noxious pain under steady state and pathology.
Supported by Damon Runyon Cancer Research Foundation
Analysis of the mechanisms that maintain intestinal regulatory T cells
Authors: Morales Elisa Cruz , Hart Andrew P , Laufer Terri M 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play an important role maintaining intestinal homeostasis. The Treg pool from the small intestine lamina propria (siLP) include two subpopulations: Helios+ Tregs, comprised of mainly thymic derived Tregs, and the microbiota-induced RORgt+ Tregs. We used K14 transgenic mice lacking peripheral TCR-MHCII to examine the phenotype and maintenance of Helios+ Tregs in siLP. We find that the siLP of K14 mice fills with Helios+ Tregs that adopt an effector phenotype independent of TCR signaling.
To dissect the mechanisms that maintain siLP Helios+ and RORgt+ siLP Treg subpopulations, we used blocking antibodies to inhibit costimulatory signals. We find that the composition of the Treg pool is dynamic; both subpopulations expand to compensate for the loss of the other to maintain intestinal Treg numbers. Moreover, we found that the balance between RORgt+ and Helios+ Tregs in siLP is differentially modulated by ICOS and CD28 signals. Thus, while Helios+ Tregs rely only on CD28 but neither ICOS nor MHCII signaling for their expansion; MHCII, CD28, and ICOS signaling play a role in RORgt+ Treg maintenance. Together, our data highlight the intricate signaling network that regulates the intestinal Treg homeostasis.
Supported by grants from: AAI Careers in Immunology Fellowship VA NIH
Parenteral vaccination strategies target protective immunity to the small and large intestine mucosa
Authors: Chen Ziqi , Tierney Flora , Raposo Bruno , Gonzalez Rodrigo , Sadeghi Ilin , Schudel Alex , Hu Quanyin , Garcia Johnny , Langer Robert , Jaklenec Ana , von Andrian Ulrich H 
Published: January 2023

Abstract
Prophylactic vaccines are most commonly given by a parenteral route. However, this strategy often confers poor protection against GI pathogens because antigens are primarily presented in peripheral lymph nodes, which do not provide the prerequisite ‘tissue imprinting signals’ to target memory T and B cells to the gut. Here, we describe a platform that enables parenteral vaccines to target protection to the GI tract. This was achieved by subcutaneous (s.c.) delivery of compounds known to imprint gut tropism onto lymphocytes. Am80, a retinoic acid receptor agonist, targeted immunity to the small intestine through up-regulation of gut-homing receptors α4β7 and CCR9, and promotion of IgA class-switching. S.c. injection of OVA + CpG combined with repeated injections of Am80 generated significantly higher and durable fecal anti-OVA IgA titers, compared to OVA + CpG without Am80. Vaccination with Am80 also increased antigen-specific CD4 and CD8 T cell trafficking to the small intestine and generated tissue resident memory T (Trm) cells. To overcome the need for multiple doses of Am80, which can cause tissue toxicity at the injection site and hinder clinical translation, we developed a polymeric nanocarrier that showed equivalent imprinting efficacy upon a single injection, lowering the cumulative dose to one-third. Furthermore, we also developed a modified vaccination strategy that could augment effector and memory T cell responses not only in the small intestine, but also in the cecum, colon and rectum, conferring protection to the entire intestinal tract. We conclude that supplementation with mucosal imprinting signals is a promising approach to enhance parenteral vaccines by generating mucosal protection against GI infections.
Supported by the Bill & Melinda Gates Foundation
TGF-β signaling is important for female reproductive tract resident memory CD8 T cell development
Authors: Hasan Mohammad H , Mithila Farha J , Pacia Jessica , Zhang Nu , Beura Lalit K 
Published: January 2023

Abstract
Tissue-resident memory T cells (TRM) patrol non-lymphoid barrier tissues and provide rapid protection against reinvading pathogens. While TRM formation in several barrier mucosal tissues including intestine and respiratory tract has been investigated, factors that control TRM establishment in the female reproductive tract (FRT) remain unclear. Here we established a model of local FRT infection, characterized CD8 TRM development and probed the role of cytokine TGF-β in TRM formation. We adopted a mouse model of intravaginal lymphocytic choriomeningitis virus (LCMV) infection that generated robust local CD8 T cell response in both the lower FRT (cervix, vagina) and upper FRT (ovaries and uterine horn). Upon viral clearance, CD8 T cells in the tissue gradually adopted mature TRM phenotype. Parabiosis confirmed most antiviral T cells in the FRT were truly resident. There were marked differences in the phenotypes of TRM in upper and lower FRT with CD103+ epithelial TRM predominating the lower FRT mucosa. Longitudinal analysis revealed TRM in both compartments undergoes attrition. The cytokine TGF-β has been implicated in TRM development in number of barrier and visceral organs. Using TGF-β RII deficient transgenic CD8 T cells, we observed that lack of TGF-β receptor signaling impairs acquisition of CD103 and CD69 expression by FRT TRM. However, TGF-β RII deficient CD8 T cells showed elevated expression of effector molecules granzyme-B and interferon-gamma upon in vivo stimulation. Taken together, our data suggest a key role of TGF-β in the formation and function of virus specific TRM in the FRT. Further investigation will provide mechanistic insight on the nature of TGF-β signaling pathway impacting FRT TRM formation.
Supported by NIH grants (P20GM121298 and P20GM109035) and Searle Scholars Foundation