bluuebunny/crossref_metadata_2025_split huggingface dataset:
Swine as a Surrogate Model for Human Brucellosis Following Oropharyngeal Vaccination with a Live Attenuated Brucella melitensis Mutant
Authors: Clapp Beata , Yang Guan , Artiaga Bianca L. , Hoffman Carol , Yang Xinghong , Driver John P , Pascual David W 
Published: January 2023

Abstract
Human brucellosis is a global health problem, and no human vaccines exist. Although Brucella infections primarily occur via the oropharynx and the upper respiratory tract mucosa, mucosal aspects of Brucella’s pathogenesis are often ignored. Mucosal vaccination with our live attenuated mutant was found to confer exceptional protection against Brucella challenge. While mice have been instrumental to study immunity to Brucella, they may be less suitable to evaluate mucosal infections. Domestic pigs’ close resemblance to human immune system and anatomical features of the oropharyngeal (OPG) mucosa enables studying mucosal brucellosis in a natural host. To assess our mutant’s immunogenicity, 8 wk-old pigs were vaccinated by directly applying 109 CFUs onto the tonsils and sublingual mucosa plus a buccal injection with 108 CFUs on days 0, 2, and 4, and study was terminated on day 60. Isolated peripheral blood mononuclear cells (PBMCs) on days 0, 15, 23, 35, and 60 as well as terminal head and neck lymphoid tissue lymphocytes were evaluated for IFN-g responses by flow cytometry. The greatest IFN-g came from CD8+ and CD4+ CD8+ T cells, as well as NK cells. Changes in peripheral blood T cells over the course of the response showed a significant reduction in the percentage of total CD4+ T cells, and a significant increase in the percentage of total CD8+ and CD4+ CD8+ T cells. Among the lymphoid tissues examined, with the exception of the tonsils where the IFN-g was mostly derived from CD8+ T cells, IFN-g came from CD4+CD8+ T cells, particularly in the mandibular lymph nodes. Thus, our Brucella mutant is immunogenic in swine and capable of eliciting elevated IFN-g responses following OPG vaccination. Work supported by R03 AI128123 & USDA-NIFA2013-01165.
Establishment of a Regulatory Microenvironment by Lactococcus Expressing Colonization Factor Antigen I (CFA/I) Fimbriae Ameliorates Type 1 Diabetes (T1D) in Non-Obese Diabetic (NOD) Mice
Authors: Nelson Andrew Scott , Maddaloni Massimo , Hoffman Carol , Abbott Jeffrey , Pascual David W 
Published: January 2023

Abstract
Oral treatment with the adhesin from enterotoxigenic E. coli, CFA/I fimbriae protect in murine models of multiple sclerosis and rheumatoid arthritis. Although CFA/I fimbriae initial mode of action is in a bystander or in an antigen (Ag)-independent fashion, protection was found to be ultimately dependent upon the induction and/or activation of auto-Ag-specific regulatory T cells (Tregs). Our recent findings show that oral dosing with a L. lactis vector expressing CFA/I fimbriae (LL-CFA/I) reduces incidence of T1D in NOD mice by 45% with a concomitant 8-fold increase in their splenic Foxp3+CD25+ Tregs. However, little is known about how protection transitions from bystander suppression to Ag-specific Tregs. We hypothesized that LL-CFA/I stimulates dendritic cells (DCs) to establish a regulatory microenvironment since DCs play an integral role in fate decisions for T cells becoming inflammatory or tolerogenic. In this study, 4 wk-old NOD mice were orally dosed with LL-CFA/I and treated every 2 wks; control groups were given L. lactis vector or PBS. At 11 wks of age, frequency of insulitis was reduced by more than half, and insulin-specific T cells were reduced in the pancreatic lymph nodes (PaLNs; P &lt; 0.05). To discern how this was mediated, groups of mice were examined at 3, 7, 14, and 21 days post-treatment for changes in DC and T cell phenotypes. As early as 3 days post-treatment, DCs exhibited significantly less expression of costimulatory molecules, CD40 and CD86, and stably maintained this phenotype for at least 7 wks. Splenic DCs showed reduced IL-6 production. These data show that orally dosing with LL-CFA/I ameliorates T1D in NOD mice by establishing a regulatory microenvironment via DCs. Work is supported by AI121745.
IL-6 producing intraepithelial lymphocytes in the colon modulate epithelial barrier integrity to protect against C. rodentium infection
Authors: Kuhn Kristi , Schulz Hanna M. , Whitney Alyssa K. , Campbell Eric L , Colgan Sean P. 
Published: January 2023

Abstract
Citrobacter rodentium infection of mice is frequently used as a model to study host responses during pathogenic gastrointestinal infection. IL-6 signaling in the colon is required for protection against C. rodentium infection; however the cellular source of IL-6 for this protection is unclear. Our previous work has identified IELs as one source of IL-6 in the colon. Therefore, we hypothesized that IL-6 produced by colonic IELs was sufficient to provide protection during C. rodentium infection. To test our hypothesis, we orally infected mice with C. rodentium following transfer of donor IL-6+/+ or IL-6−/− IELs into IL-6+/+ or IL-6−/− recipients. IL-6−/− mice that received IL-6−/− IELs had significantly more weight loss, increased intestinal histopathology, and increased bacterial translocation after 12 days of infection compared to transfer of IL-6+/+ IELs into recipient IL-6−/− mice and IL-6+/+ IELs into IL-6+/+ mice. IEL-derived IL-6 is functionally important in the maintenance of the epithelial barrier as IL-6−/− mice were noted to have increased paracellular permeability, decreased claudin-1 expression, and a thinner mucus-gel layer, all of which were reversed by transfer of IL-6+/+ IELs. In vitro studies utilizing model epithelia confirmed IL-6 was able to signal and increase claudin-1 and mucin-2 expression. Therefore, we conclude that IL-6 expression by IELs is sufficient to restore protection against C. rodentium infection through modulation of the epithelial barrier integrity.
Interleukin-33 promotes REG3γ expression in intestinal epithelial cells via activation of mTOR, STAT3 and ERK1/2
Authors: Xiao Yi , Chen Feidi , Zhao Ye , Sun Mingming , Huang Xiangsheng , Chen Liang , Yao Suxia , Liang Yuejin , Sun Jiaren , Yan Fang , Cong Yingzi 
Published: January 2023

Abstract
REG3γ, an antimicrobial peptides typically expressed by intestinal epithelial cells (IEC), plays crucial roles in maintaining intestinal mucosal barrier functions against microbiota and potentially pathogenic microorganisms in the intestines. However, the mechanisms by which regulate IEC expression of REG3γ are still not fully understood. IL-33, a member of the IL-1 cytokine family, has been implicated in maintenance of intestinal homeostasis. In this study, we investigated whether and how IL-33 regulates IEC expression of REG3γ, thus contributes to the maintenance of intestinal homeostasis. We report here that the expression of REG3γ in IEC of IL-33−/− mice is significantly lower than that in the wild type (WT) mice at both mRNA and protein levels. Treatment with IL-33 induced IEC expression of REG3γ in mouse MSIE intestinal epithelial cell line. Consistently, IL-33 also induced expression of REG3α, the human ortholog of mouse REG3γ, in human epithelial cells. Mechanistically, rIL-33 activated phosphorylation of STAT3, mTOR and ERK1/2 in MSIE cells. Knockdown of STAT3, mTOR and ERK-activating enzyme MEK by using siRNA or CRISPR abrogated IL-33 induction of REG3γ expression in IEC, indicating that IL-33 promoted IEC expression of REG3γ though activating STAT3, mTOR and ERK1/2 pathways. REG3γ has been shown to be able kill gram-positive gut bacteria. Interestingly, total gut bacteria were significantly lower in IL-33−/− mice compared to WT mice, indicating that IL-33 regulation of gut microbiota, possibly through induction of IEC REG3γ. Put all together, our study demonstrated a novel crosstalk between IL-33 and REG3γ in maintenance of intestinal homeostasis.
Antimicrobial Cytotoxic Immune Proteins in Human Milk
Authors: Filgueira Luis , Kueffer Gwendoline , Twigger Alecia-Jane , Kharoubi Solange , Walch Michael , Geddes Donna T 
Published: January 2023

Abstract
Human milk (HM) contains a large variety of immune components providing protection to the infant. The presence of the cytotoxic immune proteins perforin (Per), granulysin (Grly)and granzymes (Grzm)has not yet been shown in HM cells.
Per, Grnly and Grzm (A, B, H and M) were examined in cells isolated from samples of prepartum HM (PS) collected during pregnancy and HM collected longitudinally (2–5 sampling occasions) from mothers (n=31) during the first year postpartum, including 3 samples from breast infections. Gene expression was analyzed in RNA sequencing data and using qRT-PCR. Linear modeling and principle component analysis between the genes was conducted. Flow cytometry (n=5) was performed to investigate the presence of these immune proteins in HM cells.
Gene expression of all immune proteins was confirmed in PS and in HM during month 1 of lactation, where variation of expression was found between women and time lactation. Strong correlations were found between GrzmA – Per (r2=0.86), GrzmA – CD45 (r2=0.76), Per – CD45 (r2=0.68) and Per – GrzmB (r2=0.67). Comparison between healthy and mastitis samples showed a higher expression of CD45 and all immune peptides in mastitis. Moreover, flow cytometry analysis showed an increase in CD45 positive cells in mastitis in comparison with healthy participants (15.1% and 4.7% respectively of total cell amount).
The presence of Per, Grly and Grzm has been confirmed in HM cells. An increase in gene expression of these immune proteins has been confirmed in 3 participants suffering from breast infection. Further investigations are required to elucidate the roles of these immune peptides for the infant and/or mother
IL-10 Can Enhance FcγR-mediated Mast Cell Activation
Authors: Haque Tamara , Ryan John J 
Published: January 2023

Abstract
Mast cells are involved in many inflammatory diseases, partly by their production of cytokines that promote inflammation. IgG signaling through FcγRIII/IIb in mast cells is linked to autoimmune disease such as rheumatoid arthritis, but factors controlling this signaling remain unclear. Our group, as well as others, has previously shown that IL-10 suppresses mast cell IgE signaling, consistent with known anti-inflammatory effects of IL-10. However, we recently found that in IL-10 has unexpected pro-inflammatory effects on IgE-mediated mast cell functions. Therefore, we designed a study to determine the effects of IL-10 on FcγR signaling. Our data show that IL-10 enhances IgG-mediated IL-6 and MCP-1 production in bone marrow derived mast cells. Interestingly, we also found that IgG-induced cytokine production is suppressed by IL-10 in bone marrow derived macrophages under the same conditions. Current work is examining this different response to IL-10 to have a better understanding of the overall affect of IL-10 in the inflamed microenvironment. These studies suggest that mast cell and macrophage responses are altered by cytokines in their microenvironment, with distinct and opposite effects.
Partial protection from lupus-like disease in B-cell specific IFNAR-deficient B6.Nba2 mice
Authors: Keller Emma J , Patt Madeline M , Nemunaitis Teddy , Reth Michael , Sen Ganes C , Jorgensen Trine N 
Published: January 2023

Abstract
A majority of patients presenting with systemic lupus erythematosus (SLE) have increased serum concentrations of interferon alpha (IFNα). IFNα has previously been shown to affect lymphocyte activities such as B cell differentiation and T cell maintenance, but whether IFNα signaling specifically in B cells is required for disease pathology remains unknown. In order to examine the role of IFNα stimulation on B cells, we backcrossed mice lacking the IFNα receptor on B cells, Mb1.cre IFNARflx/flx, with the B6.Nba2 lupus mouse model. B6.Nba2.BΔIFNAR mice were evaluated alongside littermate control mice for splenomegaly, abnormal spleen subset distribution, IgG immune-complex deposition in the kidney glomeruli. Serum total IgM, total IgG and anti-chromatin IgG levels were determined monthly. BΔIFNAR mice display normal IgM and IgG levels, but decreased serum antinuclear autoantibody levels (p&lt;0.05), while deposition of IgG immune complexes in kidney glomeruli was similar to that of control mice. Both spleen weight and total splenocyte number were found to be significantly decreased in BΔIFNAR mice (p&lt;0.05). Analyses of spleen cell subsets at 4 months of age revealed a trend towards decreased plasma cells and GC B cells in spleens of BΔIFNAR mice. Despite IFNAR being removed from B cells only, we observed a shift in the balance of effector/memory to naïve CD4 T cells in BΔIFNAR mice. In summary, we found that B6.Nba2.BΔIFNAR mice were partly protected from lupus pathology including splenomegaly, autoantibody production and B cell hyperactivation. Most surprisingly, T cell activation appeared to be diminished in B6.Nba2.BΔIFNAR mice, suggesting that B cells play a central role as both APCs and ASCs in the B6.Nba2 mouse model of lupus.
RELT binds HIC and induces apoptosis by a mechanism distinct from TNFR1.
Authors: Cusick John K , Moua Pachai , Talbott George , Sumida Cara , Jacobs Aaron 
Published: January 2023

Abstract
Receptor Expressed in Lymphoid Tissues (RELT) is a human Tumor Necrosis Factor Receptor (TNFR) that is expressed most prominently in cells and tissues of the hematopoietic system. RELT has two identified homologous binding partners RELL1 and RELL2. This study sought to further elucidate the function of RELT by identifying novel protein interactions with RELT family members. Human I-mfa domain-containing protein (HIC), also known as MDFIC, was identified in a yeast two-hybrid genetic screen using RELL1 as bait. HIC is a transcription factor that was initially identified to differentially regulate HTLV and HIV gene expression. HIC is now known to regulate many other cellular processes and the hic gene is located proximally to regions of chromosome 7 (7q31.1) frequently lost in Acute Myeloid Leukemia (AML) patients. HIC physically interacts with both RELT and RELL1, as determined by in vitro co-immunoprecipitations. HIC co-localizes with RELL1 at the plasma membrane and co-localizes with RELT in intracellular compartments. Deletion mutants of RELT were utilized to determine regions of RELT required for both the activation of p38 and the induction of apoptosis in HEK-293 cells, two previously described functions of RELT. Co-immunoprecipitation experiments utilizing the deletion mutants indicate that regions of RELT proximal to the plasma membrane are sufficient for physical interaction with HIC. Overexpression of RELT induces cleavage of PARP and Caspase-3 as determined by western blotting. Interestingly, induction of apoptotic morphology by RELT overexpression was not prevented if signaling by FADD or Caspase-8 was blocked, indicating RELT induces apoptosis by a pathway distinct from death-domain containing TNFRs such as TNFR1.
Evolution of interleukin-1 receptor-like 1 and its role in rainbow trout (Oncorhynchus mykiss) resistance to Flavobacterium psychrophilum
Authors: Shaw Cassidy H. , Palti Yniv , Gao Guangtu , Wiens Gregory D 
Published: January 2023

Abstract
Rainbow trout exhibit extensive phenotypic variation in innate disease resistance and we have divergently selected lines with either increased or reduced survival following exposure to the gram-negative bacterium, Flavobacterium psychrophilum (Fp). Following five generations of selection, gene expression was analyzed between the resistant and susceptible lines by RNA-seq and RT-PCR in either whole body or spleen tissue samples. Lower base-line expression of interleukin-1 receptor-like 1 (il1rl1) was identified in susceptible line fish as well as decreased expression in response to infection. Genomic analyses identified three tandem, il1rl1 genes on chromosome 3 located within a previously identified, bacterial cold water disease QTL. Here, we report the genomic organization and evolution of the three il1rl1 genes, and develop gene specific assays to measure the expression of the individual il1rl1 genes in susceptible and resistant rainbow trout lines. Sequence homology between these three putative genes is approximately 95%. Each gene contains a signal peptide, three IG/IG-like domains, a transmembrane region and a TIR domain. Two il1rl1 genes and one pseudogene are present in Atlantic salmon (Salmo salar) genome with 90–94% sequence identity. Phylogenetic analyses suggest that the salmonid il1rl1 genes expanded by tandem duplication within the salmonid linage. Our working model is that il1rl1 gene copy number or differential expression may contribute to specific disease resistance.
Irgm1 suppresses NLRP3 Inflammasome-mediated IL-1β production
Authors: pei chunying , wang chun , xu hongwei 
Published: January 2023

Abstract
Immune-related GTPase M (Irgm1) plays a crucial role in resolving immune responses during infection. The molecular mechanism underlying the immunoregulatory function of Irgm1 is not fully understood. NLRP3 inflammasome triggered by variety of stimulus is actively involved in immune responses during both physiological and pathological conditions. In the current study, we investigate the potential role of Irgm1 in regulating NLRP3-Caspase-1-IL-1β signal cascades. To test that, bone marrow derived-macrophage and peritoneal macrophage from either Irgm1-deficient (Irgm1−/−) or wild-type (WT) mice were challenged with bacterial LPS and ATP. Secretion of IL-1β was measured by ELISA. NLRP3 Inflammasome activity was measured by ASC oligomerization, caspase-1 activity. While the expression of pro-IL-1β is not different between Irgm1-deficient (Irgm1−/−) or wild-type (WT), mature IL-1β secretion by Irgm−/− macrophage is strongly up-regulated upon stimulation as compared to macrophage from WT mouse. Furthermore, we found that the expression NLRP3, ASC and cleaved caspase-1 is also increased in the Irgm−/− macrophage. In keeping with that, ASC oligmerization as well as caspase-1 activity is significantly enhanced in the macrophage from Irgm1-deficient mice. We blocked NLRP3 inflammasome activity by using specific inhibitors against NLRP3 (MCC950) and caspase-1 (VX765). Interestingly, the addition of NLRP3 inflammasome inhibitors normalized the up-regulation of IL-1β observed in Irgm−/− macrophage. Taken together, these data suggested that Irgm1 negatively regulates NLRP3 inflammasome, which may be involved in the resolution of immune responses.
A novel method for the detection of IFN-lambda 3 binding to cells to quantify IFN-lambda receptor expression on epithelial and immune cell subsets
Authors: Santer Deanna , Minty Gillian , Mohamed Adil , Baldwin Lesley , Bhat Rakesh , Joyce Michael , Egli Adrian , Tyrrell D. Lorne , Houghton Michael 
Published: January 2023

Abstract
Type III interferons (IFN-lambdas) are important antiviral cytokines that also modulate immune responses by acting through a unique IFN-λR1/IL-10R2 heterodimeric receptor. Conflicting data has been reported for which human cells express the IFN-λR1 subunit and directly respond to IFN-λ. Since the commercially available anti-IFN-λR1 flow cytometry antibodies we tried were suboptimal, we developed a novel method to measure IFN-λ3 binding to IFN-λR1/IL-10R2 on the surface of cells via flow cytometry. We found that Huh7.5 hepatoma cells bound IFN-λ3 to the greatest extent with the lowest Kd(app) (83.2nM), and had the corresponding highest induction of IFN stimulated genes (ISGs). Raji and Jurkat cell lines, representing B and T cells, respectively, moderately bound IFN-λ3 and had lower ISG responses. U937 cells, representing monocytes, did not bind IFN-λ3 and therefore, did not have detectable ISG induction. We confirmed that IFN-λ3 was bound to the surface of cells through imaging flow cytometry. Importantly, lentivirus shRNA knockdown of IFNLR1 in Huh7.5 cells decreased our binding signal proportionally and reduced ISG induction by up to 93%. IFN-λ3 responsiveness increased over time with maximal ISG responses seen at 24 hrs for all but one gene. We next applied our assay to human peripheral blood mononuclear cells and saw that only specific immune cell subsets bound IFN-λ3 (eg. plasmacytoid dendritic cells and B cells). These data confirm our new IFN-λ3 binding assay can be used to quantify where the IFN-λ receptor is expressed and reflects IFN-λ3 responsiveness. Knowing which cells express the IFN-λ receptor will be crucial for determining how IFN-λ3 modulates the adaptive immune response.
Recombinant Fasciola hepatica fatty acid binding protein down regulates the expression of inflammatory cytokines from macrophages induced by LPS in vitro
Authors: Ramos-Benítez Marcos Javier , Espino Ana M. 
Published: January 2023

Abstract
Gram-negative sepsis develops as a consequence of an uncontrolled bacterial infection that constantly release LPS into the bloodstream triggering an unrestrained systemic inflammatory response that leads to multi-organ failure and death. Molecules that can neutralize inflammation induced by LPS are highly pursued by the scientific community. Our research group revealed that a 12kDa F. hepatica FABP (Fh12) exhibit potent anti-inflammatory properties and prevented the septic shock development in mice. Recently, we optimized the expression and purification of a recombinant variant of Fh12, termed Fh15 and demonstrated that both molecules possess immunological identity and similar capacity to inhibit the NF-κB activation induced by LPS in THP1-CD14 cells. In the current work we investigated whether Fh15 can suppress the expression of inflammatory cytokines from murine bone marrow-derived macrophages (BMDM) and also examined whether the integrity of the secondary structure is determinant for the anti-inflammatory role of this molecule. qRT-PCR results demonstrated that Fh15 significantly suppressed the expression of TNF-α and IL-1β cytokines (p&lt;0.0041) from BMDM in response to LPS. Moreover, Fh15 significantly repressed the expression of CD14 co-receptor (p&lt;0.0145), reinforcing our notion that one of mechanisms used by Fh15 to suppress the inflammatory response is by targeting this co-receptor. Additionally, anti-Fh12 antibodies does not inhibit the capacity of Fh15 to block the NF-κB activation in THP1-CD14 cells. After a thermal treatment, Fh15 capacity to suppress NF-κB activation was not reduced. Circular dichroism analysis confirmed that the secondary structure of Fh15 was not altered by thermal denaturation.
Metformin Suppresses LPS-induced CXCL8 Expression
Authors: Xiao Zhihui , Wu Wenjun , Poltoratsky Vladimir 
Published: January 2023

Abstract
Human chemokine CXCL8 plays an important role in cancer development by promoting tumor growth, angiogenesis, and tumor cell migration. It also recruits monocytes that, after differentiation into tumor associated macrophages, further increase tumor progression. Anti-diabetic therapy with Metformin reduces cancer incidence and mortality and has a potential application in anti-cancer therapies. In this study we tested the effects of metformin on LPS-induced CXCL8 expression, cell proliferation, and migration. We used the modified HEK293 cell line which express the Toll like receptor 4 signaling pathway receptor complex, but lack expression of CXCL8 receptors.
We demonstrated that Metformin significantly suppresses the LPS-induced CXCL8 expression in a dose-dependent manner. Metformin reduced LPS-induced CXCL8 activation on a transcriptional level through the suppression of NF-κB, but not AP-1 or C/EBP transcription factor activities. Immunofluorescent analysis revealed that Metformin suppresses the NF-κB translocation into the nucleus. In addition, Metformin suppresses LPS-independent HEK293/TLR4 cell migration, but not cell proliferation. In conclusion, our data suggested that metformin suppresses CXCL8 expression via inhibiting NF-κB activities, and inhibits HEK293/TLR4 cell migration without affecting cell proliferation.