bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of the interactions between fish serum proteins and pathogenic bacteria by using label-free quantitative proteomics
Authors: Dong Miao , Tang Sze Wing , Shen Wei , Au Doris W.T. , Lam Yun Wah 
Published: January 2023

Abstract
Fish blood is a valuable source of antimicrobial proteins which form a crucial part of the innate immune system, but the types and functions of these proteins are still not fully understood. The objectives of this study are to systematically characterize the serum proteins from fish that stably bind to bacteria and to investigate how bacteria respond to these interactions. Serum from turbot, but not the heat-inactivated control, significantly decreased the number of E. tarda bacteria. By conjugating fish serum proteins with fluorescent dye, we showed that E.tarda were coated with multiple labeled proteins. In order to systematically identify these bacteria-binding proteins, we used E.tarda cells to capture turbot serum proteins and subjected the samples to shotgun proteomic analysis, followed by label-free quantitation. A total of 76 fish proteins have been identified, including known antimicrobial proteins such as complement components and Wap65-2. 34 proteins with no previously known immunological function were also identified to bind to E.tarda. This approach also allowed the study of the proteomic changes in E.tarda exposed to turbot serum. Our data indicated a rapid decrease of translation factors and an upregulation of membrane components in response to serum treatment. Interestingly, fish serum induced a rapid suppression of the expression of bacterial antioxidant enzymes like catalase and led to an accumulation of reactive oxygen species, suggesting that fish serum can override a bacterial self-defence mechanism. Taken together, this work offers a comprehensive view of the interactions between fish serum proteins and pathogenic bacteria, and reveals previously unknown factors and mechanisms in fish innate immunity.
Expanded CD4+ CD25− FoxP3+ Regulatory T cell population in HIV-TB patients
Authors: Quiroga M Florencia , Vecchione Maria Belen , Suarez Guadalupe , Laufer Natalia , Ben Graciela , Sued Omar , Angerami Matias T 
Published: January 2023

Abstract
Tuberculosis (TB) and HIV alter the immune system, and coinfected (HIV-TB) individuals usually show deregulations in lymphocyte’s immune functions. We previously observed an increased frequency of “unconventional” Treg (uTreg, CD4+CD25−FoxP3+) during HIV-TB. Therefore, we aimed to explore the phenotype and function of uTreg and conventional Treg subsets (cTreg, CD4+CD25+FoxP3+) among HIV-TB (n=34) patients and healthy donors (HD, n=30). We evaluated the expression of CD39, PD1, GITR and maturation status by flow cytometry in cTreg and uTreg. Also, IL-10, TGF-β, IFN-γ production and suppressive capacity were analyzed in cocultures of Tregs and effector lymphocytes. We found diminished expression of CD39 and higher levels of PD-1 on uTreg compared to cTreg in both HIV-TB (p&lt;0.01) and HD (p&lt;0.001). Nevertheless, GITR expression was increased only in uTreg of HIV-TB patients (p&gt;0.05). In addition, uTreg and cTreg showed differences in maturation status both in HIV-TB and HD groups (p&lt;0.05, p&lt;0.01 respectively), due to Effector Memory uTregs expansion (p&gt;0.05). Interestingly, both HIV-TB and HD showed higher production of IFN-γ only in uTreg population (p&lt;0.01), though no significant differences were observed for IL-10 and TGF-β production between uTreg and cTreg. Moreover, IFN-γ+ cells were restricted to the CD39− uTreg population. Finally, when suppressive capacity was evaluated, both uTreg and cTreg inhibited T cell-proliferation and IFN-γ production in a similar extent (p&lt;0.05). These findings suggest that uTregs, which are expanded during HIV-TB coinfection, are fully functional in terms of inhibition of T cell responses despite of an altered surface expression of characteristic Treg markers and cytokine production.
Environmental bioaerosol antigen Methanosphaera stadtmanae induces a TH17 inflammatory lung response via TLR4 signaling
Authors: Bernatchez Emilie , Gold Matthew J , Langlois Anick , Blais-Lecours Pascale , Duchaine Caroline , Marsolais David , McNagny Kelly M , Blanchet Marie-Renée 
Published: January 2023

Abstract
Bioaerosols in occupational environments are highly associated with the development of inflammatory lung diseases. The archaea specie Methanosphaera stadtmanae (MSS) is found in high concentrations in poultries, dairy farms and swine confinement buildings bioaerosols (up to 108 archaea/m3). MSS induces a strong specie-dependent inflammatory lung response in mice, characterized by T cells, eosinophils, neutrophils, and IgG production. However, the polarity of lung response induced by MSS and the mechanisms underlying this inflammatory response remain unknown. Using a mouse model, we show that MSS induced a weak TH2 (CD4+/IL-13+T cells) strong TH17 (CD4+/IL-17A+T cells) lung response, characterized by IgG1 (but not IgG2a and IgE) production. Moreover, mice did not develop airway hyperresponsiveness following MSS exposure. Interestingly, increasing MSS quantity led to a lower eosinophil count associated with a decreased TH2 response. Using transgenic mice, we found that eosinophils, mast cells and ILC2 cells are not required for the TH2 and TH17 inflammatory response to MSS. However, Tlr4−/− mice (but not Tlr2−/−) had reduced airway inflammation compared to WT mice after exposure to MSS, indicating a crucial role for TLR4 activation in this specific response. Finally, heat denaturation and zymography studies suggested TLR4 activation likely occurs through MSS protein antigen recognition rather than by enzymatic activity and cell damage. We conclude that MSS induces a TH17 inflammatory lung response via TLR4 signaling.
How genetically modified probiotic bacteria can modify human dendritic cell functions?
Authors: Tóth Márta 
Published: January 2023

Abstract
Selected lactic-acid bacterial strains such as Lactobacilli are classified as probiotics with beneficial effects on the human host. It has previously been shown that the group of Lactobacillus rhamnosus/casei/paracasei are able to prevent cytokine-induced apoptosis of intestinal epithelial cells by secreted factors identified as peptidoglycan-hydrolases. To exert their protective and immunomodulatory effects, these bacteria have to interact with the underlying epithelial and immune cells, such as dendritic cells (DCs). DCs recognize Lactobacilli and their components by pattern recognition receptors (PRR) that may induce stimulatory or inhibitory T-lymphocyte responses.
Our results showed that the inflammatory response of human monocyte-derived DCs (moDCs) to bacterial stimuli and the ability to polarize T-lymphocytes to different directions was highly dependent on the intact structure of the bacterial cell wall, while the expression of the cell surface molecules of moDCs was independent on the modification of the peptidoglycan (PG) layer. It was also found that the secreted bacterial cell wall preparations were able to activate moDCs demonstrated by the increased levels of pro – and anti-inflammatory cytokines and elevated expression of cell surface markers independently on the genetic manipulation of the cell wall.
Based on these results we conclude that the genetic modification of the bacterial cell wall is able to modulate the stimulatory potential of moDCs and has an impact on the outcome of T-lymphocyte responses at the level of the whole bacteria. In contrast to the effects of the specific PG extracts, the functions of moDCs are independent on the genetically modified bacteria or their cell wall and its components.
Type I IFN dependent and independent mechanisms of protection in C. difficile infection
Authors: Snyder Deann Teresa , Hedges Jodi F , Robison Amanda , Broadaway Susan C. , Walk Seth T. , Jutila Mark A 
Published: January 2023

Abstract
Clostridium difficile infection (CDI) is increasingly prevalent in individuals in health care settings and in the community. C. difficile is a gram positive, anaerobic bacteria that forms spores resistant to common disinfectants. Upon infection, C. difficile spores germinate, divide and produce damaging toxins resulting in intestinal inflammation. While inflammatory mediators are known to affect disease severity, the role of type I IFN signaling in CDI has not been characterized. We hypothesized that type I IFN signaling was important for innate protection in CDI. IFNAR-deficient mice succumbed to CDI more rapidly than wild type mice. Since type I IFNs can reduce inflammasome activity, we assessed IL-1β expression in multiple tissues, and discovered that IFNAR mice expressed more IL-1β protein, and less insulin-like growth factor (IGF) transcripts in response to CDI. To investigate a potential treatment, we treated mice with the type I IFN-inducing dsRNA mimetic, polyIC. PolyIC treatment delayed onset of symptoms in survival studies. Surprisingly, the protective effect of polyIC was not a result of IFNAR signaling; treatment increased IGF and decreased IL-1β in both wild type and IFNAR-deficient mice. Finally, treating wild type mice with recombinant type I IFN also reduced disease severity. These findings suggest direct type I IFN signaling protects from CDI and treatment with polyIC may protect mice via a pathway independent of IFNAR signaling. Considering that recombinant type I IFN is already used clinically and novel treatment options for CDI are needed, further investigation of innate immune pathways may present new treatment options for this emerging infection.
Increased Treg cells and expression of CD39/CD73 during listeriosis in aging mice
Authors: Cavanaugh Christopher , Gaines Dennis , Pereira Marion , Williams Kristina , Alam M. Samiul 
Published: January 2023

Abstract
Foodborne Listeria monocytogenes (LM) is a cause of serious illness & death in the elderly. Aging is associated with increased susceptibility to pathogens. We investigated the cause of this increased susceptibility of aging mice to LM infection using an oral model. Young & old C57BL/6 mice were gavaged with murinized LM and T-helper (Th) cell responses were evaluated. After infection, tissues were examined for bacterial burden & histology. Isolated splenocytes were analyzed for intracellular Th1/Th2 cytokine expression (TNF-α, IL-10 and IFN-γ) & distribution of Treg (CD4+CD25h) & Th17 cells before & after infection. The expression of adenosine producing enzymes, CD39/CD73, in Th cells was analyzed by FACS & qRT-PCR. Old LM-infected mice lost more body-weight & had a higher bacterial burden in the liver/spleen than young-infected mice. Uninfected old mice showed a higher baseline proinflammatory response when compared to young control mice. After infection, the proinflammatory cytokine, IFN-g, was higher in young mice as compared to old mice. An increase in Th-17 cells was observed in the older mice before & after infection. Increased levels of IL-10 & Treg cells were observed in older mice after infection. Higher expression of CD39/CD73 was observed in the Th cells of the older mice & the Treg expression of CD39 doubled after infection, suggesting increased production of adenosine, an anti-inflammatory mediator. Increased LM infection in aging mice is associated with reduced IFN-g response & increased anti-inflammatory responses from Treg cells with high expression of CD39 in splenocytes. Increased accumulation of splenic Th17 cells in LM-infected old mice can further contribute to increased inflammation leading to LM pathogenesis.
Host response to Neisseria gonorrhoeae infection in female wild type mice is dictated by the dominant sex hormone
Authors: Francis Ian P , Shaik-Dasthagirisaheb Yazdan , Islam Epshita , Gray-Owen Scott D , Wetzler Lee 
Published: January 2023

Abstract

BACKGROUND
Gonorrhea, a sexually transmitted disease (STD) caused by Neisseria gonorrhoeae (GC), causes major sequelae in women. Epidemiology suggests a link between symptomatic gonorrhea and the normal female hormone cycle. The aim of this study was to evaluate the murine gonorrhea-model immune responses to infection during progesterone dominate diestrus, and estrogen dominate estrus.

METHODS
Hormone cycle stage was determined by cytology of vaginal washes. Mice were infected with 107 bacteria in PBS directly placed into a uterine horn. Serum, upper and lower genitourinary (UGU, LGU respectively) tract tissues were collected at 6 or 18 h post infection. Cytokines levels in these tissues were measured by multiplex immuno assay. Gene transcription in UGU and LGU samples was measured by microarray.

RESULTS
Infection during estrus induced a moderate cytokine response at the site of infection. Cytokine levels were much greater when infection occurred during diestrus. Microarray analysis of infected diestrus tissues showed significant enrichment of those pathways associated with host immune response when infection occurred in diestrus.

CONCLUSIONS
GC infection during diestrus was shown to be highly inflammatory. Immune pathways were strongly upregulated as determined by Gene set enrichment analysis (GSEA) of microarray data. The most differentially expressed pathways were associated with pattern-recognition-receptor activation and cytokine response. GSEA analysis of estrus tissue showed very limited enrichment of any immune processes. These data together lead us to conclude that the inflammatory response to GC infection in mice is strongly dependent on the dominant sex hormone at the time of infection.
Gene expression profiles predict treatment outcomes in Brucellosis
Authors: Dufort Matthew J , Hanniffy Sean , Álvarez Raquel Conde , Gorvel Vilma Arce , Altman Matthew C. , Khaenam Prasong , Solera Javier , Gersuk Vivian H. , Bosilkovski Mile , Moriyón Ignacio , Linsley Peter S , Gorvel Jean-Pierre 
Published: January 2023

Abstract
Brucellosis is a zoonotic disease caused by Brucella bacteria, acquired by humans through contact with or consumption of products from infected animals. Treatment outcomes vary widely; some patients recover, others relapse, and others develop chronic symptoms despite therapy. We employed expression profiling using RNA sequencing (RNAseq) to better understand variation in outcomes and investigate disease mechanisms. We performed RNAseq analysis of whole blood samples from 125 Brucellosis patients and 51 healthy controls from Macedonia. Patients were subdivided based on disease history into primary, secondary, and chronic infection. Secondary and chronic cases did not show differences in gene expression from healthy controls or from each other. Primary cases displayed numerous transcriptional changes relative to healthy controls. Up-regulated genes were enriched for response to interferon-gamma, cytolysis, T cell proliferation, and cell cycle; down-regulated genes, for B cell proliferation. Primary cases were further subdivided based on outcome into those who resolved with treatment and those who developed disease relapse during treatment. In comparison to healthy controls, relapse cases showed larger magnitude differences in expression at diagnosis than resolution cases; this suggests that relapse could potentially be predicted based on transcription levels at time of diagnosis. Differences in gene expression observed at diagnosis were no longer present 6–9 months after diagnosis. Our results indicate opportunities to prognostically identify patients that may require more intensive treatment and monitoring, and to better understand the symptomology of and immune response to Brucella infection.
CD4+ T cells from human neonates and infants respond variously against antigens of Staphylococcus aureus, but not against Bifidobacter longum ssp. infantis
Authors: Brunner-Weinzierl Monika C , Pierau Mandy , Schlueter Dirk , Lampe Karin , Arens Christoph , Jorch Gerhard , Katrin Vogel 
Published: January 2023

Abstract
T cells in human neonates, infants, and adults differ dramatically in the initiation, strength, and stability of their responses. In this study, we investigate cellular mechanisms of CD4+ T cells from neonates, infants and adults to show the antigen specific response to Staphylococcus aureus and Bifidobacterium longum ssp. infantis.
T cells from surgically excised adenoids of infants in different ages, cord blood and peripheral blood from healthy donors were characterized by flow cytometry and functional assays. Therefore CD14+ monocytes were incubated with extracts of Staphylococcus aureus and Bifidobacterium longum ssp. infantis to stimulate enriched T cells.
CD4+ CD45RA and CD4+ CD45R0 T cells proliferate and up regulate the activation-associated molecule CD25 in response to Staphylococcus aureus whereas stimulation with Bifidobacterium longum ssp. infantis had no effect. In functional assays, we could show that T-cell proliferation is induced by Staphylococcus aureus via TCR-specific signal transduction. In this line, the antigen-specific response can be reduced by blockade of MHC class II molecules. Surprisingly, an inverse correlation between the percentages of Staphylococcus aureus induced proliferating T cells and age of infants is observed.
In the case of activation with Staphylococcus aureus functional specific T cells are identified in cord blood, adenoids of infants, and adults with age-dependent characteristics.
This work highlights the gap between specific T cell responses of neonates, infants, and adults in terms of quality and quantity. Our findings will help to understand the relationship between pathogens and T cells and to optimize future strategies for therapeutic interventions.
Towards an Understanding of Protective Cellular Immunity - a TCR Transgenic Mouse Reactive with Chlamydia muridarum
Authors: Poston Taylor , Qu Yanyan , Russell Ali , Girardi Jenna , Nagarajan Uma , Darville Toni 
Published: January 2023

Abstract
A major limitation in understanding the development of protective T cell memory against Chlamydia is the difficulty in characterizing low frequencies of antigen-specific T cells. To better understand the kinetics and phenotype of specific CD4 T cells, we developed a C. muridarum-specific T-Cell Receptor (TCR) transgenic (Tg) mouse. Primed T cells from C57BL/6 mice immune to C. muridarum were stimulated with EB/RB for 5 days and fused with BW5147 cells. Specific clones producing IL-2 and IFN-γ were harvested and screened for TCR Vα and Vβ by qPCR. Cloned transgenes were extracted, purified, and co-microinjected into fertilized oocytes. PCR and FACS confirmed integration and expression. Tg T cells were analyzed ex vivo for activation markers and cytokine production after EB/RB stimulation. 90% of peripheral CD4+ T cells express TCR transgenes and exhibited enhanced proliferation and increased expression of IFN-γ, IL-2, and CD69, compared to controls. Naive T cells were adoptively transferred from CD45.2 Tg mice into CD45.1 mice one day before intravaginal inoculation with C. muridarum. Tg cells showed enhanced proliferation and TNF-α, IL-2, and IFN-γ production by day 5 compared to controls. Tg T cells collected from SLOs and oviducts on days 5, 8, 12, 22, 44, and 65 post-infection exhibited enhanced expression of memory and activation markers. Adoptive transfer of Tg or WT CD4 T cells to Rag1−/− mice rescued an otherwise lethal C. muridarum genital infection. WT and Tg CD4 T cells exhibited comparable chlamydial clearance from the genital tract. Adoptive transfer of Chlamydia-specific CD4+ T cells offers a powerful approach to characterize cellular activation, differentiation, and memory development.
Development of a metal-based detection method for simultaneous protein and gene expression analysis in single cells by mass cytometry
Authors: Mavropoulos Anastasia , Majonis Daniel , He Mingxiao , Park Emily , Ornatsky Olga 
Published: January 2023

Abstract
The Fluidigm® CyTOF system is a mass cytometer that uniquely enables high-dimensional single-cell analysis of complex populations. Mass cytometry is based on inductively coupled plasma time-of-flight mass spectrometry used for multiplex proteomic analysis. In this approach, metal-conjugated affinity reagents are used to tag the components of cells. The cells are nebulized and sent to an argon plasma, ionizing the multi-atom metal tags, which are then analyzed by a time-of-flight mass spectrometer.
Gene expression can be finely tuned through the synthesis of RNA and through the control of its stability and location. Misregulation of gene expression can have severe consequences, such as developmental disorders, cancer and autoimmune diseases. The ability to acquire complete spatial-temporal profiles of gene expression is therefore critical for the understanding of disease pathophysiology, medical diagnostics and drug discovery.
In this presentation, we discuss the development of a multiplex method for targeted RNA detection using the Fluidigm CyTOF® and Advanced Cell Diagnostics RNAscope® platforms. This novel assay includes the hybridization of RNA-specific target probes, followed by signal amplification and ending with the binding of amplifier-specific metal-labelled probes. Presently, we are able to detect four different mRNA probes in a single-cell analysis setting. The detection of RNA is compatible with Fluidigm products such as Maxpar® antibodies. Future developments will include high-dimensional detection that enables researchers to investigate the simultaneous expression profile of RNA and protein across millions of cells.
Transcutaneous delivery of protein tyrosine phosphatase ameliorates inflammatory skin diseases
Authors: Kim Won-Ju , Koo Ja-Hyun , Cho Hyun-Jung , Kim Ji-Yun , Lee Sohee , Kim Jong Hoon , Suh Minah , Shin Eui-Cheol , Choi Je-Min 
Published: January 2023

Abstract
Transcutaneous delivery of therapeutic drugs has many advantages of topical treatment over the systemic administration for inflammatory skin diseases such as atopic dermatitis or psoriasis. However, the greatest challenge for transcutaneous drug delivery limits clinical applications due to the skin tissue barrier. Here, we identified a novel transcutaneous delivery peptide, AP, from human neuronal adhesion protein (Astrotactin 1) which could deliver a macromolecule such as a protein into skin tissue. AP-conjugated EGFP protein exhibited significantly higher intracellular transduction efficacy in HaCaT (keratinocyte), NIH3T3 (fibroblast) and Jurkat (T cell) cells compared with control proteins. In addition, AP-EGFP and –dTomato protein was efficiently localized into the dermis as well as epidermis in mouse skin tissue following transcutaneous administration. Next, we generated AP-rPTP protein, which is chimeric protein of AP and phosphatase domain of TC-PTP. AP-rPTP inhibited phosphorylation of STAT1, 3, 5 in mouse splenocytes and T cells and reduced cytokines production by activated splenocyets. To confirm its in vivo relevance, we transcutaneously applied it to dermatitis mouse models. Multiple skin patch administrations of AP-rPTP protein significantly ameliorated tissue inflammation in oxazolone-induced contact dermatitis and imiquimod-induced psoriasis mouse model. Our results collectively demonstrate that AP is a novel human derived transcutaneous delivery peptide and AP-rPTP protein could be a therapeutic biomolecule in inflammatory skin diseases such as allergic dermatitis and psoriasis.