bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bovine colostrum increases the viability and the healing process of IPEC-J2 intestinal porcine epithelial cells while modulating the barrier function and Escherichia coli-mediated inflammatory responses.
Authors: Bouchard Michaël , Blais Mylène , Deschenes Karine , Robitaille Gilles , Arcand Yves , Asselin Claude , Lessard Martin 
Published: January 2023

Abstract
Piglet gut response to weaning results in impaired intestinal barrier function, increased inflammatory response and susceptibility to enteric diseases. As the use of antibiotics as growth promoters will be restricted in the swine industry in Canada, novel feeding strategies to improve gut health are needed. This study aims to determine the potential of defatted bovine colostrum (BC) and its fractions, the serocolostrum (SC) and caseins (CAS), to promote viability and the healing process and to modulate barrier integrity and the inflammatory response induced by enterotoxigenic E. coli (ETEC) in IPEC-J2 intestinal epithelial porcine cells. To evaluate cell viability and wound healing, XTT cell proliferation and migration scratch assays were performed on IPEC-J2 cells incubated with BC, SC, or CAS. Monolayer permeability of ETEC infected IPEC-J2 cells incubated with BC, SC or CAS was monitored by transepithelial electrical resistance measurements. Inflammatory and oxidative responses were assessed by gene expression analysis of several cytokines and chemokines by qPCR. Incubation with BC derivatives led to 8 to 14% increases IPEC-J2 cell viability, in comparison to untreated cells (P ≤ 0.05). IPEC-J2 cell migration was increased by 42% after SC addition, as opposed to non-treated cells (P ≤ 0.05). BC and SC treatments significantly reduced ETEC-mediated increases in IL-8 and CCL20 expression (P ≤ 0.05). BC treatment also significantly reduced IPEC-J2 cell monolayer permeability induction after ETEC infection. These results show that BC and SC fractions improve intestinal epithelial cell integrity and decrease ETEC-mediated inflammatory responses. Therefore, BC should be considered as a potential alternative to antibiotics.
AlgR controls in vivo persistence of Pseudomonas aeruginosa
Authors: Penaranda Cristina , Hung Deborah 
Published: January 2023

Abstract
Pseudomonas aeruginosa (PsA) is one of the most virulent opportunistic human pathogens and is often highly resistant to antibiotic therapy. Although it is traditionally considered an extracellular pathogen, PsA has been shown to invade and persist inside mammalian cells. We hypothesize that this ability to persist intracellularly allows bacteria to become antibiotic insensitive, leading to the formation of reservoirs for recurrent infections. We have developed an in vitro infection model where we identified that the two-component response regulator AlgR is required for infection of human bladder epithelial cells. We demonstrate that AlgR controls invasion, via pilus expression, and intracellular persistence, via an unknown mechanism. Furthermore, we show that although its cognate histidine kinase, AlgZ, is also required, AlgR phosphorylation is not required for persistence. RNA profiling of host cells infected with WT PsA shows induction of hallmark TNFα signaling via NFKβ, which is absent in cells infected with ΔalgR PsA, suggesting that recognition of infecting bacteria may be necessary for their intracellular persistence. Finally, we show that AlgR is also required in an in vivo model of murine urinary tract infection, demonstrating that the in vitro model correlates with chronic infections in a whole animal model. Our results advance our understanding of the requirements for in vivo bacterial persistence and identify a potential target for the development of novel antibiotics.
Chlamydia-Specific CD8+ T cells Enhance Neutrophil Infiltration into Chlamydia-Infected Female Mouse Genital Tract
Authors: Li weidang , Ramsey Kyle H. , Murthy Ashlesh K. 
Published: January 2023

Abstract
We recently demonstrated that Chlamydia-specific TNF-α producing and TNF receptor 2 bearing CD8+ T cells mediate chlamydial pathogenesis in the mouse female upper genital tract (UGT). Additionally, TNF-α production from CD8+ T lymphocytes is necessary for pathogenesis; however, CD8+ T lymphocytes are not a sufficient source of TNF-α production for pathogenesis. Myeloid cells including neutrophils are important sources of TNF-α production and have been implicated in chlamydial pathogenesis. Herein, we evaluated frequency of neutrophils in Chlamydia muridarum-infected female genital tracts of C57BL/6J (WT) mice, mice deficient in Chlamydia-specific CD8+ T cells (OT-1 mice) and TNFR2 knockout (KO) mice. Frequencies of Gr-1+ cells (neutrophils) in UGT were evaluated on days 4, 9, 15, and 24 following genital Chlamydia muridarum infection. OT-1 and TNFR2 KO displayed significant reduction in neutrophil frequency compared to WT animals on day 9 following chlamydial inoculation. Furthermore, we found that OT-1 mice replete with WT CD8+ T cells at the time of chlamydial inoculation display restoration of neutrophil (Ly-6G) frequency to WT levels. These results suggest that Chlamydia-specific CD8+ T cells engage other inflammatory cells such as neutrophils in infected genital tracts in order to cause chronic pathologies in the upper reproductive tract.
Role of Interferon-γ in promoting disease severity in neonatal Bordetella pertussis infection
Authors: Mitchell Ashley E. , Scanlon Karen , Carbonetti Nicholas 
Published: January 2023

Abstract
Pertussis is an infectious disease caused by the bacterial pathogen Bordetella pertussis. In recent years, there has been a re-emergence of pertussis disease. Pertussis morbidity is extensive and spans across all age groups, but a higher incidence of severe disease and mortality is exhibited in young infants. Mortality is associated with severe respiratory infection and other systemic secondary co-morbidities. The infant immune system has reduced capacity to generate proinflammatory/T-helper (Th) 1 cell polarizing responses. Secretion of Interferon gamma (IFN-γ), a Th1 polarizing cytokine, by immune cells plays a critical role in macrophage activation and microbicidal activity in the lung. Furthermore, B. pertussis respiratory challenge of adult IFN-γ receptor deficient mice results in systemic dissemination and lethality. We hypothesize that the dissemination and lethality observed in our neonatal mouse model is a result of inadequate IFN-γ signaling or production, which contribute to insufficient control of the infection. We found that B. pertussis infection results in a significantly lower level of transcription of IFN-γ in the lungs and increased bacterial dissemination in neonatal mice when compared to adult mice. Additionally, infected neonatal mice deficient in IL-10, a potent regulator of IFN-γ, showed reduced dissemination and significantly increased IFN-γ and other pro-inflammatory cytokines as compared to age-matched wild type mice. Future studies include characterization of IFN-γ secreting immune cells during infection. Results generated by this work will contribute to a better understanding of severe disease in infants, addressing the limitations of infant immunity and treatment options for pertussis.
Dynamics and function of group 1 CD1-restricted T cells during disseminated Staphylococcus aureus infection
Authors: Visvabharathy Lavanya , Wang Chyung-Ru 
Published: January 2023

Abstract
Staphylococcus aureus (SA) is frequently associated with nosocomial and community-acquired infections. Some promising vaccine candidates act by priming conventional T cell responses, but less is known about the contribution of non-conventional T cells to SA infection. Group 1 CD1-restricted T cells are a unique subset of T cells that respond to lipid antigens presented by group 1 CD1 (CD1a, -b, –c) molecules. These molecules have low polymorphism in their antigen-binding grooves, making them attractive vaccine targets. Previous studies have shown that group 1 CD1-restricted T cells respond to Mycobacterium tuberculosis (Mtb) lipids. As group 1 CD1 molecules can accommodate a variety of lipid species, we questioned whether and how group 1 CD1-restricted T cells can be activated by SA lipids.
Using a transgenic mouse strain expressing human group 1 CD1 molecules (hCD1Tg), we found that group 1 CD1-restricted T cells recognize SA lipid during primary infection. hCD1Tg mice were also protected against SA kidney pathology compared with WT control mice at late times post-infection. Group 1 CD1-restricted T cells produced both IL-17 and IFN-γ in response to SA lipid. Fractionation of SA lipid into 11 discrete parts found that portions enriched in cardiolipins and phosphatidyl glycerol contained immunodominant group 1 CD1-restricted antigens. Finally, group 1 CD1-restricted SA lipid-specific T cells can be specifically detected via FACS to ultimately assay differences in gene expression between these and conventional SA Ag-specific T cells. These findings suggest a role for CD1-restricted, SA lipid-specific T cells in control of systemic SA infection, with strong implications for vaccine design.
Increasing Domain Coverage Improves Neutralizing Potency of C. difficile Toxin-specific Antibodies
Authors: Li Lu , Cole Leah E. , Jetley Utsav , Zhang Jinrong , Pacheco Kristl , Ma Fuqin , Zhang Jianxin , Mundle Sophia , Yan Yanhua , Barone Lucianna , Rogers Christopher , Beltraminelli Nicola , Quemeneur Laurence , Kleanthous Harry , Anderson Stephen F. , Anosova Natalie G. 
Published: January 2023

Abstract
Clostridiumdifficile (C.difficile)is a significant human pathogen. C.difficile infection (CDI) causesclinical symptoms ranging from diarrhea to life-threatening fulminant pseudo membranous colitis. The pathogenesis of C. difficile is mediated by two large exotoxins, toxins A and B. These two toxins are highly homologous, single chain proteins consisting of four functional domains: N-terminal glucosyl transferase domain (GTD), cysteine protease domain (CPD), translocation domain (TLD) and a C-terminal receptor-binding domain (RBD). The important role played by anti-toxin sera and antibodies in the prevention of primary and recurrent CDI has been described and demonstrated in both clinical and pre-clinical studies; however, work focused on the impact of the toxin domain specificity of these antibodies is limited. To address this deficit, sera from a C. difficile vaccine immunized hamsters and toxin-specific human monoclonal antibodies (mAbs) were used to assess the impact of toxin domain specificity on antibody mediated inhibition of cytotoxicity in a Vero cell-based functional assay. Results from toxin domain immunoabsorption assays using hamster anti-toxinsera indicated that no single domain fragment from either toxin A or toxin B could inhibit neutralizing activities. Anti-toxin A activity was prevented with a combination of GTD and CTD fragments while anti-toxin B activity required the GTD, CTD and CPD fragments to block activity. Assays with human mAbs demonstrated that combining mAbs that target multiple toxin domains greatly improves neutralizing potency when compared to equivalent concentrations of either a single mAb or a combination of mAbs against a single domain.
A novel function of ADP-ribosyl cyclases to regulate the length of the small intestine revealed in CD38/CD157 double knockout mice
Authors: ISHIHARA Katsuhiko , YAHAGI Ayano , ISEKI Masanori 
Published: January 2023

Abstract
Mammalian ADP-ribosyl cyclase includes two ectoenzymes; GPI-anchored, Bone marrow stromal cell antigen-1 (BST-1)/CD157 and a type II membrane protein, CD38. Although they are expressed in various tissues including the lung, kidney, intestine, brain, spleen and bone marrow, the enzymatic activity of CD38 is dominant at physiological pH. We generated and analyzed CD157 deficient mice (CD157KO), and reported that CD157 is a positive regulator of humoral immunity in 129 x C57BL/6J (B6) mixed background. Recently, various groups reported novel functions of murine CD157 in intestinal regeneration and depressive or anxious behaviors. Thus, CD157 is now appreciated as an entero-neuro-immune regulator. The phenotypes of CD38KO also revealed the multiple functions of CD38 in immunity, metabolism, social behaviors, as well as in the kidney and lung.
To further clarify the function of ADP-ribosyl cyclase family, we generated CD38/CD157 doubly deficient mice (CD38/157DKO) by sequential targeting at ES cell level, and compared with CD38KO, CD157KO and wild type in B6 background. A novel phenotype unique to B6.CD157KO was enhanced humoral immune responses to TNP-LPS, indicating the suppressive role of CD157 in TLR signal on marginal zone B cell. Although CD38/157DKO developed apparently normal with no histological changes in the major tissues by HE staining, the length of the small intestine increased by approximately 20% at 4 months old. Among the lymphoid organs, the cell numbers in the mesenteric lymph node increased by 1.5 times without changes in the lineage or subset, probably reflecting elongation of the small intestine.
These data suggest that ADP-ribosyl cyclases, CD38 and CD157, have roles in regulating functions of the small intestine.
Differences in psychosocial behavior, salivary IgA, and oral microbiota in student-athletes and sedentary college students.
Authors: Costan Sophia , Appasamy Pierette M. , Edenborn Sherie , Ubinger Elizabeth , Carrillo Andres 
Published: January 2023

Abstract
Identification of behaviors and physiology that positively impact psychosocial and physical well-being in college students is very important. Exercise and social connectedness have been shown to influence psychosocial well-being, immunologic function, and the microbiome. Our previous studies demonstrated reduced anxiety, enhanced oral immunity and an altered oral microbiota in student athletes during the “in-season” period (Lamb et al, 2017). In our present studies, we found that the the composition of the oral microbiota in female NCAA Division III ice hockey players was different in athletes in the “off-season” compared to “in-season.” Specifically, there was a greater proportion of streptococcal strains in the “off-season” athletes, whereas behavioral measures, including anxiety and perceived stress, and salivary cortisol levels, were similar. Salivary IgA levels were different between the two groups. In order to examine whether participation on an athletic team alone is beneficial to students with respect to behavioral and immunologic measures, our most recent study compared female student athletes (NCAA Division III soccer players), “in-season”, with female college students who were not athletes. As expected, student athletes were more “fit”. We found that the degree of perceived stress and depression was higher in the non-athletes, although both groups reported similar degrees of social connectedness. Non-athletes reported a higher rate of illness compared to the athletes. Saliva samples were collected and are being evaluated for microbial composition, IgA and cortisol levels, in order to further define relationships between psychosocial and physical well-being, the microbiome and the immune response.
miR-146a modulates IgA class-switch DNA recombination by targeting Smad2, Smad3 and Smad4 genes
Authors: Daw Cassidy C. , Zan Hong , Casali Paolo 
Published: January 2023

Abstract
IgA is the predominant antibody isotype at mucosal surfaces. It plays a critical role in mucosal homeostasis and provides a first line of immune protection. Class-switch DNA recombination (CSR) to IgA is a strictly controlled process that involves cytokine induction of transcription factors and expression induced cytidine deaminase (AID). CSR to IgA is initiated by TGF-β activation of Smad2, Smad3, Smad4, and Runx3, which bind to the Iα promotor region to activate germline Iα-Cα transcription. This critically opens up the Igα locus and is greatly enhanced by retinoic acid (RA). As we have suggested, CSR is modulated by epigenetic modifications, including microRNA-mediated gene regulation. miR-146a has been shown to regulate T cell, dendritic cell and macrophage functions in innate and adaptive immune responses. Its role, however, in B cells and in the antibody response, is unknown. Here we showed that, in B cells, miR-146a expression is downregulated by LPS/CD154, TGF-β, IL-4, anti-BCR plus RA, the stimuli that induce a robust CSR to IgA. Deficiency of miR-146a in miR-146a−/− B cells resulted in upregulated Smad2, Smad3 and Smad4 expression, increased recruitment of Smad2, Smad3 and Smad4 to Ia promoter, as well as elevated germline Iα-Cα transcription and CSR to IgA, without a significant alteration in AID expression and CSR to other isotypes, including IgG1. Indeed, as we also showed, miR-146a targets the 3′UTRs of Smad2, Smad3 and Smad4 mRNAs, thereby inhibiting expression of these genes. Thus, by downregulating the level of Smad2, Smad3 and Smad4 in B cells, miR-146a plays an important role in the modulation of the IgA response.
Erythroid suppressor cells promote feto-maternal tolerance through arginase-2 and PDL-1
Authors: Elahi Shokrollah , Delyea Cole , Bozorgmehr Najmeh , Dunsmore Garett , Shahbaz Shima , Huang Vivian 
Published: January 2023

Abstract
Survival of the allogenic pregnancy depends on the maintenance of immune tolerance to paternal alloantigen at the feto-maternal interface. Multiple localized mechanisms contribute to fetal evasion from the mother’s immune rejection as fetus is exposed to a wide range of stimulatory substances such as maternal allo-antigens, microbes and amniotic fluids. Here, we demonstrate that CD71+ erythroid cells are expanded at the feto-maternal interface and in the periphery during pregnancy. These cells exhibit immunosuppressive property and their abundance is associated with a Th2 skewed immune response, as their depletion results in a pro-inflammatory immune response at the feto-maternal interface. In addition to their function in suppressing pro-inflammatory responses in vitro, maternal CD71+ erythroid cells inhibit an aggressive allogeneic response directed against the fetus such as reduction in TNF-a production through arginase-2 activity and PD-1/PDL-1 interactions. Their depletion leads to the failure of gestation due to immunological rejection of the fetus. Similarly, fetal liver CD71+ erythroid cells exhibit immunosuppressive activity. Therefore, immunosuppression mediated by CD71+ erythroid cells on both sides (mother/fetus) is crucial for feto-maternal tolerance. Thus, our results reveal a previously unappreciated role for CD71+ erythroid cells in pregnancy and indicate that these cells mediate homeostatic immunosuppressive/immune-regulatory responses during pregnancy.
Unique Cytokine Profile of Novel Innate Lymphoid Cells in Human Decidua
Authors: Vazquez Jessica , Li Yan , Stanic-Kostic Aleksandar 
Published: January 2023

Abstract
Immune cells at the maternal-fetal interface play a complex role in regulation of vascular remodeling, fetal tolerance and protection from infection. We have previously presented the identification of two novel CD56highNKs based on Eomes expression (Eomeslo/hi). Activation of these subsets revealed preferential production of IL-8 and IL-17A by decidual innate lymphoid cells (ILCs).
Decidual specimens were dissected from term placentas and mononuclear cells (MCs) were isolated by mechanical (GentleMACS) and enzymatic (Collagenase, DNAse) disruption. MCs were then simulated with PMA/ionomycin or IL-23, then labeled by flurochrome-conjugate antibodies against surface CD3, 14, 16, 19, 34, 45, 49a, 56, 94, 117, 127, 335 and intracellular Granzyme B, IL-8, IL-17A, IL-22, INFγ, TNFα. Data acquisition was performed using BD Fortessa flow cytometer in a 5 laser, 18 detector configuration and data analysis was performed using FlowJo 10.2.
Activation of decidual ILCs revealed a unique milieu of cytokine production. The novel subsets, CD56high Eomeslo/hi, preferentially produced IL-8 and IL-17A, while decreasing production of INFγ, Granzyme B, and TNFα. In addition, as expected, ILC3s and LTi-like cells produced IL-17A and IL-22. Furthermore, differences in response were observed between decidual basalis and decidual parietalis ILCs, with decidual parietalis ILCs being more responsive to stimulation.
Preliminary results suggest that both CD56high Eomeslo/hi subsets demonstrate a similar altered cytokine profile that is conducive to decidual remodeling. Further gene expression analysis will reveal if these novel subsets represent a truly unique population or are members of the expanding tissue-resident NK cell family.
Novel strategies to assess the synaptic interface of primary human T cells from peripheral blood and lymphoid tissue
Authors: Steblyanko Maria A. , Anikeeva Nadia , Buggert Marcus , Betts Michael R. , Sykulev Yuri 
Published: January 2023

Abstract
Our current understanding of the kinetics and structural features of T cell synaptic interfaces has largely been determined through the use of glass supported artificial bilayers and in vitro-derived T-cell clones or lines. How these findings apply to the primary human T cells isolated from blood or lymphoid tissues in unclear, given the significant difficulty in obtaining sufficient amounts of these cells for use on conventional glass-supported planar bilayers. Moreover, emerging data suggest that tissue resident T cells and T cells from lymphoid organs are significantly different in their phenotype and functional activity from those in peripheral blood, further solidifying the need to better understand the features of T cell synaptic interfaces in primary human T cells. To this end, we developed a new mini scale approach exploiting bilayers built into multichannel flow slides that allows us to perform imaging studies of T cell/bilayer interfaces with as few as 105 cells primary T cells isolated from human peripheral blood and lymphoid tissues without sacrificing the quality of the analysis. Our results indicate that while primary peripheral blood CD8+ T cells can share synaptic interface properties similar to in vitro derived T cell clones and lines, we found significant differences in the structure of the synaptic interface formed by primary lymphoid tissue compared to peripheral blood T cells, consistent with different efficiencies of these T cells.
A novel peptide pool with broad infectious antigen and MHC coverage for use as a positive stimulation control or as a means to elicit general T-cell responsiveness
Authors: Castro Aaron , Holenya Pavlo , Eckey Maren , Schulz Marco , Wenschuh Holger , Reimer Ulf , Tech Tatjana , Chan Kam , Janani Ramesh , Broaten Blake , Kern Florian 
Published: January 2023

Abstract
T-cell activation assays such as Elispot or Intracellular Cytokine Staining (ICS) require appropriate positive stimulation controls. Compared with conventional assay controls based on polyclonal stimulation (e.g. PHA/Ionomycin), peptide pools have the advantage of providing a more physiological, TCR-mediated signal. A frequently used positive control is the so-called ‘CEF’ pool, containing 23 selected class-I-MHC-presented T-cell-stimulating peptides from CMV, EBV, and Influenza A. As a result of frequent and widespread exposure, most if not all individuals have T-cells specific for at least one of these agents. However, the ‘CEF pool’ only covers the most frequent HLA alleles in European-Caucasoids but not other ethnic groups where its usefulness is limited. We, therefore, designed an extended pool referred to as ‘CEFX pool’ with significantly improved coverage of both infectious agents and HLA-alleles (176 peptides), which also contains numerous CD4 T-cell-stimulating peptides. This novel pool provides a much stronger and more universal positive control than the ‘CEF’ pool and was successfully tested in various assay formats including Elispot, ICS, and ELISA of stimulated T-cell supernatants. For specific questions, sub-pools for CD4 or CD8 T-cell stimulation or not containing CMV-derived peptides are available. Because of its broad antigen coverage we are currently exploring the novel pool’s potential for measuring/monitoring general T-cell responsiveness in clinical situations such as immunodeficiency, immunosuppression, or immunosenescence.