bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mycobacterium tuberculosis extracellular vesicles induce the formation of granulomalike structures in an in vitro model
Authors: Wong-Baeza Isabel , Paredes Víctor Gabriel García , Silva-Miranda Mayra , Ruiz-Sánchez Bibiana Patricia , Castañeda-Casimiro Jessica , Hernández-Solis Alejandro , Cicero-Sabido Raúl , Serafín-López Jeanet , Estrada-Parra Sergio , Estrada-García Iris 
Published: January 2023

Abstract
Tuberculosis (TB) is an airborne disease caused by Mycobacterium tuberculosis. Only 10% of the infected individuals develop an active disease, while 90% of the infected individuals contain the mycobacteria inside structures known as granulomas, in a latent, asymptomatic state. The lung granuloma is formed by a core of infected macrophages, foam cells and epithelioid macrophages, surrounded by several lymphocyte subsets (CD4 T cells, CD8 T cells, Tγδ cells, NK cells, and B cells). M. tuberculosis releases extracellular vesicles, both spontaneously and under stress conditions; these vesicles carry components, derived from the mycobacterial cell wall, that are ligands for human Toll-like receptors (TLR) 2/6 and 4; as a result, these vesicles activate the innate immune system. To determine if M. tuberculosis extracellular vesicles induce the formation of a granuloma-like structure, we used an in vitro granuloma model, with peripheral blood mononuclear cells from individuals with latent TB (positive for the Quantiferon test). We found that M. tuberculosis extracellular vesicles induced the formation of granuloma-like structures that were similar in numbers, diameters and cellular composition (percentage of monocytes, T cells, Tγδ cells, NK cells and ILCs) to the granuloma-like structures formed in response to live M. tuberculosis H37Rv. These results suggest that M. tuberculosis extracellular vesicles participate in the cellular activation and migration processes that lead to granuloma formation in the lungs of TB patients.
Supported by a grant 20190106 from Secretaría de Investigación y Posgrado, Instituto Politécnico Nacional.
Microbiota metabolites SCFA stimulate epithelial migration to promote wound healing through MFGE8 and PAK1
Authors: Bilotta Anthony James , Ma Chunyan , Huang Xiangsheng , Yang Wenjing , Yao Suxia , Cong Yingzi 
Published: January 2023

Abstract
How microbiota regulate intestinal homeostasis is still not completely understood. Emerging evidence indicates that gut microbiota metabolites play crucial roles in maintenance of intestinal homeostasis and regulation of Inflammatory Bowel Disease (IBD). Many patients with IBD are deficient in the microbial metabolites of dietary fibers, short chain fatty acids (SCFAs). To date, the evidence for use of SCFAs as a treatment for IBD has been inconsistent, highlighting the need for further investigation. In this study, we demonstrate that SCFAs promote intestinal epithelial cell (IEC) wound healing. Treatment of mouse and human IEC in vitro with the SCFAs acetate, propionate, and butyrate enhanced wound closure by promoting cell migration, which was independent of cell proliferation. Further analysis of random cell migration revealed that SCFAs increased cell speed, cell persistence, and consequently mean squared displacement. To investigate the mechanism of enhanced migration, we performed microarray of mouse-derived enteroids, which identified p21 activated kinase 1 (PAK1) and milk fat globule-EGF factor 8 (MFGE8), two genes implicated in tissue repair and motility, to be highly upregulated. We confirmed upregulation of MFGE8 and PAK1 via qRT-PCR and western blot. Inhibition of PAK1 and CRISPR knockout of MFGE8 abrogated the effects of SCFAs on migration. Lastly, the above effects were dependent upon SCFAs inhibition of histone deacetylases (HDAC), as treatment of IECs with the HDAC inhibitor, trichostatin A, fully recapitulated the effects of SCFAs on migration and expression of MFGE8 and PAK1. These findings elucidate a pathway in which SCFAs inhibition of histone deacetylases may enhance wound healing in IBD.
Intestinal IL-17R signaling controls liver inflammation by constraining microbiome induced TLR9 signaling and IL-18 production
Authors: Castillo Patricia , Kumar Pawan , Hand Timothy W , Kolls Jay K 
Published: January 2023

Abstract
Th17 cells and the intestinal microbiome have been linked to liver diseases such as autoimmune and viral hepatitis. Patients exhibit intestinal dysbiosis and increased bacterial translocation. In a mouse model of T cell mediated liver inflammation, concanavalin A (Con A) hepatitis, global Il17ra deficiency and gentamicin treatment are protective. Yet, it is unknown if IL-17 contributes to microbial dysbiosis and bacterial translocation in hepatitis, and how these factors together affect disease.
To study this, we generated intestinal epithelial-specific IL-17RA deficient mice (Il17rafl/flx villin cre+ mice). Absence of enteric IL-17R signaling induced commensal dysbiosis and increased intestinal Th17 cells and Il18. These mice were more susceptible to Con A, exhibiting worsened liver damage and lethality. Single cell RNA sequencing partnered with protein-based assays revealed elevated liver IL-18, IFNγ, and FasL in Il17rafl/flx villin cre+ mice, while cohousing and antibiotic studies implicated the microbiome in disease exacerbation. Il17rafl/flx villin cre+ mice also had increased liver CpG levels, which induced IL-18. Moreover, anti-IL18 ameliorated Con A hepatitis.
We propose that disrupted enteric IL-17 signaling promotes intestinal dysbiosis and translocation of CpG to the liver, stimulating TLR9 and IL-18 production. IL-18 then induces hepatic lymphocyte activation and cell death to worsen inflammation. Understanding the mechanism underlying exacerbated disease in Il17rafl/flx villin cre+ mice will elucidate the role of enteric Th17 and the microbiome in hepatitis, with broader implications on the effects of impaired intestinal immunity and subsequent release of microbial products seen in other diseases.
Sensing Fungal Dysbiosis by Gut-Resident CX3CR1+ Mononuclear Phagocytes Aggravates Allergic Airway Disease
Authors: Li Xin , Leonardi Irina , Semon Alexa , Doron Itai , Gao Iris H , Putzel Gregory Garbe’s , Kim Youngjun , Kabata Hiroki , Artis David , Fiers William D , Ramer-Tait Amanda E , ILIEV ILIYAN 
Published: January 2023

Abstract
Sensing of the gut microbiota, including fungi, regulates mucosal immunity. Whether fungal sensing in the gut can influence immunity at other body sites is unknown. Here we show that fluconazole-induced gut fungal dysbiosis has persistent effects on allergic airway disease in a house dust mite challenge model. Mice with a defined community of bacteria, but lacking intestinal fungi were not susceptible to fluconazole-induced dysbiosis, while colonization with a fungal mixture recapitulated the detrimental effects. Gut-resident mononuclear phagocytes (MNPs) expressing the fractalkine receptor CX3CR1 were essential for the effect of gut fungal dysbiosis on peripheral immunity. However, how mycobiota influence immunity in gut distal sites is not well understood. We developed protocols for gut-targeted depletion of phagocytes to investigate the influence of fungi on gut-lung crosstalk. Depletion of CX3CR1+ MNPs or selective inhibition of Syk signaling downstream of fungal sensing in these cells ameliorated lung allergy. These results indicate that disruption of intestinal fungal communities can mediate gut-lung-directed immune crosstalk and aggravate disease severity through fungal sensing by gut-resident CX3CR1+ MNPs.

Highlights
Gut fungal dysbiosis persistently aggravates allergic airway disease (AAD) in mice.
Gut colonization by commensal fungi is both required and sufficient to aggravate AAD in a mycobiome-free mouse model.
Intestinal CX3CR1+ mononuclear phagocytes (MNPs) are essential for the systemic effects of gut fungal dysbiosis on AAD.
Inhibition of Syk-mediated fungal sensing in intestinal CX3CR1+ MNPs ameliorates AAD.
IL-17 receptor and IL-22 receptor signaling in Citrrobacter rodentium infection
Authors: Matsunaga Yasuka , Wanek Alanna , Song Kejin , Flemington Cathy , Clark Trevon , Bitoun Jacob , Kolls Jay 
Published: January 2023

Abstract
Th17 cells producing cytokines, IL-17A and IL-22 are important key player in host defense. We have reported that intestinal epithelial expression of IL-17 receptors in gut epithelial cells control the development of microbiota. To study the roles of these receptors in response to an attaching and effacing pathogen we challenged Citobacter rodenium (C. rodentium) to intestinal epithelial specific IL-17RA or IL-22RA1 KO mice. Conditional deletion of Il17ra or Il22ra1 resulted in increased bacterial growth in the colon as well as enhanced dissemination to the spleen. It suggested both cytokine receptors are required for control of this infection. In addition this was supported by unbiased RNAseq analysis of the colonic epithelium that showed diminished expression of Pigr, Duox2 and Duoxa2 in the absence of IL-17RA signaling and reduced Bcl3 expression in the absence of IL-22ra1 expression. To elucidate whether production from epithelial cells effect bacteria killing we cultured crypt and stimulated IL-17A. It resulted in increasing Pigr, Duoxa2 and Duox2 were associated with increasing bacteria killing. Pigr binds dimeric IgA and translocation into lumen from lamina propria, while Duox2 and Duoxa2 make complex and produce hydrogen peroxide into lumen from apical epithelial cell membrane. These result suggested mucosal immunity to C. rodentium requires both IL-17RA and IL-22ra1 signaling and that IL-17RA regulates transcytosis of C. rodentium specific IgA as well as luminal hydrogen peroxide concentrations to control bacterial growth. In contrast, Il-22Ra1 regulates epithelial Bcl3 expression, which restrains TNFα and NF-κB signaling in the colonic epithelium.
Maternal Immunoglobulin A protects against the development of necrotizing enterocolitis in preterm infants
Authors: Hand Timothy W , Gopalakrishna Kathyayini , Macadangdang Benjamin R , Rogers Matthew , Tometich Justin T , Ji Junyi , Firek Brian A , Baker Robyn , Burr Ansen H.P. , Ma Congrong , Good Misty , Morowitz Michael 
Published: January 2023

Abstract
Neonates are particularly susceptible to infection by colonizing microbes, and mammals protect their offspring via antibodies secreted into breast milk. Necrotizing Enterocolitis(NEC) is a disease of preterm infants characterized by intestinal epithelial damage and inflammation associated with the microbiota. The incidence of NEC is significantly lower in infants fed with maternal milk, though the mechanisms underlying this protective benefit are not clear. Here, we show that maternal Immunoglobulin A (IgA) is an important factor in protection against NEC. Analysis of IgA-binding within fecal samples from preterm infants indicated that breast milk was the predominant source of IgA in the first month of life and that a relative decrease in the fraction of bacteria bound by IgA is associated with the development of NEC. Sequencing of IgA-bound and unbound bacteria revealed that NEC was associated with increasing domination of the IgA unbound microbiota by Enterobacteriaceae, prior to disease onset. Further, we confirmed that IgA is critical in preventing NEC in a murine model, where we demonstrate that pups reared by IgA deficient mothers are susceptible to disease despite exposure to maternal milk. This study illustrates the importance of maternal IgA in shaping the host-microbiota relationship of preterm neonates and provides evidence that IgA is a critical factor in maternal milk necessary for the prevention of NEC.
Indole-3-carbinol prevents murine colitis development via an IL-22-dependent mechanism that regulates anti-microbial peptides and mucus production
Authors: Busbee Philip Brandon , Alrafas Haider , Dopkins Nicholas , Nagarkatti Mitzi , Nagarkatti Prakash S 
Published: January 2023

Abstract
Colitis is an inflammatory bowel disease characterized by acute or chronic inflammation within the colon. In our previous work, we showed that indole-3-carbinol (I3C), a naturally-occurring plant product found in a number of cruciferous vegetables, was able to prevent development of colitis in murine models. I3C prevented colitis-associated microbial dysbiosis characterized by an overabundance of potentially pathogenic gram-negative bacteria (e.g. Bacteroides acidifaciens), in addition to increasing anti-inflammatory gram-positive butyrate-producing species (e.g. Roseburia). This increase in butyrate by I3C, validated by metabolomics analysis, leads to a decrease in the pro-inflammatory T cell response (Th17) and an increase in anti-inflammatory Tregs, which was confirmed by sodium butyrate supplementation experiments. In the current study, we further defined the mechanisms by which I3C was able to prevent colitis-associated microbial dysbiosis and dysregulation of gut microbial metabolome by evaluating expression of key intestinal regulatory genes in immune cells and colonic epithelial cells (CECs). Transcriptome analysis and PCR validation of CECs showed mucus production (e.g. Muc2) and colonic anti-microbial peptides (e.g. beta-defensins) were significantly increased after I3C. I3C was also shown to, in a butyrate-independent manner, increase expression of colonic bacterial defensive interleukin-22 (IL-22) by epigenetic regulation. Lastly, in vivo IL-22 neutralization experiments negated the beneficial effects of I3C, thus proving this cytokine was crucial in I3C-mediated prevention of colitis development and regulation of key intestinal regulatory components such as mucus and AMP production.
B cells produce IL-27 to upregulate PD-L1 expression and promote breast cancer development
Authors: Yan Hui , Viswanadhapalli Suryavathi , Chupp Daniel , Fernandez Maria , Wu Shuai , Wang Jingwei , Moroney Justin , Taylor Julia , John IM , Rivera Carlos , Luo Yiliao , Liu Junhao , Sareddy Gangadhara , Casali Paolo , Vadlamudi Ratna , Xu Zhenming 
Published: January 2023

Abstract
The host immune response can be corrupted by cancer cells during tumor progression. Here we showed that B lymphocytes played an important role in promoting murine breast cancer (BCa) development, as B cell-deficient mMT mice displayed virtually abrogated tumor growth from transplanted syngeneic ERa+ breast adenocarcinoma cells, concomitant with reduced PD-L1 expression and increased BCa cell death. In murine BCa tumors and tumor-associated lymph nodes, B cells were the major source of IL-27, a pleotropic cytokine with both pro- and anti-inflammatory functions. Deficiency in IL-27 expression specifically in B cells abolished the BCa tumor growth, showing that the effect of B cells in BCa development was mainly through IL-27 production, as likely induced by TLR ligands in the tumor microenvironment together with CD154 and IL-21 expressed by tumor-infiltrating CD4+ T cells. IL-27 treatment of BCa cells resulted in STAT1 and STAT3 phosphorylation, upregulated PD-L1 gene expression and enhanced cell growth. In addition, IL-27 induced activated B cells to express PD-L1 in vitro, consistent with high levels of PD-L1 expression by B cells in tumors. In humans, IL-27 receptor genes were highly expressed in the breast tissue, at levels comparable to those in lymphoid organs, and high IL-27 gene expression was associated with reduced survival in BCa patients, who displayed increased circulating IL-27 levels. Finally, IL-27 supported human BCa cell growth in the presence of tamoxifen, suggesting a role of IL-27 in the drug resistance. Thus, TLR-fueled IL-27 induction in B cells reinforces PD-L1 expression in the tumor environment to drive BCa cell growth and to promote generation of PD-L1hi B regulatory cells, leading to BCa tumor progression.
Precisely timed histone deacetylase inhibition creates a highly proliferative intratumoral CD8 T cell population and sensitizes tumors to checkpoint blockade
Authors: McCaw Tyler R , Liu Mingyong , Li Mei , Starenki Dmytro , Cooper Sara , Arend Rebecca , Forero Andres , Buchsbaum Donald , Randall Troy D 
Published: January 2023

Abstract
We previously showed that a class I histone deacetylase (HDAC) inhibitor, entinostat (ENT), dramatically improves CD8 T cell-mediated control of TS/A murine mammary tumors, but only when given in a precise window. If given too early ENT halts initial T cell activation and expansion and has no effect if given too late— once T cells start becoming dysfunctional. This led us to hypothesize that HDAC inhibition can maintain CD8 T cells in a less differentiated state that enhances effector and proliferative capacities. To test this, we first treated TS/A tumor-bearing mice with ENT and performed single cell RNA-sequencing on sorted tumor-infiltrating CD8 T cells. Relative to vehicle, ENT treated samples clustered separately and included a distinct, highly proliferative cluster of CD8 T cells, a finding corroborated by bulk RNA-sequencing and flow cytometry. Although TS/A tumors do not respond to anti-PD1 alone, ENT-induced changes in CD8 T cell phenotype sensitized tumors to PD1 blockade, leading to tumor rejection in most mice when both agents were timed appropriately. Patient tumors, however, have often passed this window to intervene; thus, we devised a tripartite strategy to “reset the clock”. Treating first with an oncolytic virus to liberate new antigens and stimulate new tumor-specific CD8 T cells, then with ENT and anti-PD1 at the right times significantly impaired growth of late-stage tumors and even led to rejection. Our findings show that precisely timed HDAC inhibition generates highly proliferative and strongly cytotoxic CD8 T cell populations whose functions can be further elaborated with correctly timed checkpoint blockade. Resetting the clock using a tumor-lytic strategy then renders this a potent immunotherapy platform.
Differences in T cell responses to Bordetella Pertussis in adults as a function of whole cell versus acellular childhood vaccination
Authors: da Silva Antunes Ricardo Filipe , Babor Mariana , Pomaznoy Mikhail , Khalil Natalie , Cortese Mario , Mentzer Alexander J , Seumois Gregory , Petro Christopher D , Purcell Lisa A , Vijayanand Pandurangan , Crotty Shane , Pulendran Bali , Peters Bjoern , Sette Alessandro 
Published: January 2023

Abstract
In the mid-1990s vaccine-related side effects prompted the replacement of the whole Pertussis (wP) by a new acellular Pertussis vaccine (aP). Unexpectedly, whooping cough cases have recently increased, and the switch to the aP vaccine suspected to underlie this rise in morbidity, despite the use of routine boosters. Here we compared the immune responses to aP boosters in children who received their initial doses with either wP or aP vaccines. The use of ex vivo AIM assays highlighted a Th2 vs Th1/Th17 differential polarization of PT-specific memory CD4+ T cells as a function of childhood vaccination. The nature of the differences was further investigated for memory subset composition, activation, exhaustion marker expression, and transcriptomic profiles. Remarkably, donors originally primed with aP were defective in their ex vivo capacity to expand memory cells following a booster aP immunization and less capable to proliferate in vitro in response to PT epitopes. By contrast antibody responses are boosted in both aP and wP cohorts following IgG subclass distribution. Most recently, we have assessed early immune responses followed aP boosting for the aP and wP cohorts using longitudinal transcriptomics data from PBMC. We have identified several modules of genes that are co-expressed; using cell frequency data from CyTOF we infer modules correlated with specific cell types. Furthermore, we identified certain modules that show differential expression profiles between the two cohorts and are mostly enriched in immune related functions. In conclusion, our data suggest that the original priming after birth with aP and wP vaccines induce different T cell phenotypes associated with long-term T cell polarization.
Differential infection of dendritic cell subsets by virus-based vaccine platforms
Authors: Chiale Carolina , Robek Michael D 
Published: January 2023

Abstract
The use of virus-based vectors as vaccine platforms represents an attractive alternative to immunization with non-replicating antigens. However, the precise mechanism by which many virus-based vaccine vectors prime the immune response remains unknown. Specific dendritic cell (DC) subtypes play key roles in priming immune responses and controlling virus replication, but their functions in generating protective immunity following vaccination with viral vectors are not well understood. We hypothesized that highly immunogenic viral vectors based on alphaviruses and rhabdoviruses infect DCs and promote direct antigen presentation that leads to the efficient induction of antigen-specific T cells. We found that distinct DC subtypes are differentially infected with specific virus-based vaccine vectors, and display differences in activation following infection. Mouse bone marrow-derived DCs that are produced by GM-CSF stimulation and resemble moDCs rapidly succumbed to infection. In contrast, DCs that are produced by Flt3L stimulation and resemble cDCs and pDCs were efficiently infected but survived. DC activation following virus infection was more pronounced in the Flt3L-derived DCs, and showed distinctive requirements for further virus replication. Although Flt3L-derived DCs produced significant amounts of type I IFN when infected with the vaccine platforms, the mechanism of DC infection and activation was independent of type I IFN receptor signaling. These results highlight the differential survival and activation of specific DC subsets following infection with viral vaccine vectors, and indicate potentially unique roles of DC subtypes in activating the immune response following immunization.
Regulatory Cell Therapy for Sjögren’s Syndrome with E. coli Colonization Factor Antigen I (CFA/I) Fimbriae
Authors: Akgul Ali , Nelson Andrew S , Maddaloni Massimo , Hoffman Carol , Ella Bhagyaraj , Abbott Jeffrey R , Nguyen Cuong , Pascual David W 
Published: January 2023

Abstract
Sjögren’s syndrome (SjS) is one of the most common autoimmune diseases impacting several million people every year in the US. SjS results from autoimmune attack of exocrine (salivary and lacrimal) glands, and women are nine times more likely to be affected than men. To date, no vaccine or therapeutic exists to cure SjS, and patients must rely on lifelong therapies to treat symptoms. Oral treatment with the adhesin from enterotoxigenic E. coli CFA/I fimbriae has been shown to protect against several autoimmune diseases in an antigen (Ag)-independent manner. Lactococcus lactis was recently adapted to express CFA/I (LL-CFA/I), and found effective in suppressing inflammation by the induction of infectious tolerance via Ag-specific regulatory T cells (Tregs) producing IL-10 and TGF-β. We hypothesized that in the spontaneous, genetically defined model for SjS in C57BL/6.NOD-Aec1Aec2 mice, oral treatments with LL-CFA/I would offset the development of inflammatory T cells via Treg induction. To test this, 6 wk-old C57BL/6.NOD-Aec1Aec2 mice were orally dosed with LL-CFA/I and treated every 3 wks; control groups were given L. lactis vector or PBS. We found a significant recovery in salivary flow following treatment with LL-CFA/I by 28 wks of age. Treatment also significantly reduced the incidence of inflammatory infiltrates into the submandibular and lacrimal glands compared to PBS-treated mice. Treg analysis revealed a significant 4-fold increase in Foxp3+ Tregs expressing CD25, not CD39, and produced a 3-fold increase in IL-10. These data show that orally dosing with LL-CFA/I weakens or halts SjS progression via activation of Tregs. These studies will provide the basis for future testing in SjS patients. Work supported by NIH DE026450.