454 results on '"Clostridium Infections immunology"'
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2. Monoclonal antibody-mediated neutralization of Clostridioides difficile toxin does not diminish induction of the protective innate immune response to infection.
- Author
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Denny JE, Alam MZ, Mdluli NV, Maslanka JR, Lieberman LA, and Abt MC
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- Animals, Mice, Enterotoxins immunology, Disease Models, Animal, Antibodies, Neutralizing immunology, Bacterial Proteins immunology, Female, Cytokines metabolism, Broadly Neutralizing Antibodies immunology, Mice, Inbred C57BL, Bacterial Toxins immunology, Antibodies, Monoclonal immunology, Antibodies, Monoclonal pharmacology, Clostridioides difficile immunology, Immunity, Innate, Clostridium Infections immunology, Clostridium Infections prevention & control, Clostridium Infections microbiology
- Abstract
Clostridioides difficile infection causes pathology that ranges in severity from diarrhea to pseudomembranous colitis. Toxin A and Toxin B are the two primary virulence factors secreted by C. difficile that drive disease severity. The toxins damage intestinal epithelial cells leading to a loss of barrier integrity and induction of a proinflammatory host response. Monoclonal antibodies (mAbs) that neutralize Toxin A and Toxin B, actoxumab and bezlotoxumab, respectively, significantly reduce disease severity in a murine model of C. difficile infection. However, the impact of toxin neutralization on the induction and quality of the innate immune response following infection is unknown. The goal of this study was to define the quality of the host innate immune response in the context of anti-toxin mAbs therapy. At day 2 post-infection, C. difficile-infected, mAbs-treated mice had significantly less disease compared to isotype-treated mice despite remaining colonized with C. difficile. C. difficile-infected mAbs-treated mice still exhibited marked neutrophil infiltration and induction of a subset of proinflammatory cytokines within the intestinal lamina propria following infection that is comparable to isotype-treated mice. Furthermore, both mAbs and isotype-treated mice had an increase in IL-22-producing ILCs in the intestine following infection. MAbs-treated mice exhibited increased infiltration of eosinophils in the intestinal lamina propria, which has been previously reported to promote a protective host response following C. difficile infection. These findings show that activation of host protective mechanisms remain intact in the context of monoclonal antibody-mediated toxin neutralization., Competing Interests: Declaration of competing interest Linda A. Lieberman is an employee of Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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3. Screening and selection of eubiotic compounds possessing immunomodulatory and anti-Clostridium perfringens properties.
- Author
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John FA, Gaghan C, Liu J, Wolfenden R, and Kulkarni RR
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- Animals, Cell Line, Macrophages drug effects, Macrophages immunology, Immunologic Factors pharmacology, Immunomodulating Agents pharmacology, Immunomodulating Agents chemistry, Poultry Diseases microbiology, Poultry Diseases immunology, Anti-Bacterial Agents pharmacology, Clostridium Infections veterinary, Clostridium Infections immunology, Chickens, Clostridium perfringens physiology, Clostridium perfringens drug effects
- Abstract
Eubiotics are water and/or feed additives used in poultry to promote gut health and control enteric burden of pathogens, including Clostridium perfringens. While several eubiotic compounds (ECs) are being introduced commercially, it is essential to devise an in vitro model to screen these compounds to assess their immunomodulatory and antimicrobial properties prior to their testing in vivo. A chicken macrophage cell-line (MQ-NCSU) was used to develop an in vitro model to screen the immunological and anti-C. perfringens properties of 10 ECs: monobutyrin, monolaurin, calcium butyrate, tributyrin, carvacrol, curcumin, green tea extract, rosemary extract, monomyristate, and tartaric acid. An optimal concentration for each EC was selected by measuring the effect on viability of MQ-NCSU cells. Cells were then treated with ECs for 6, 12, and 24 h. and expression of interferon-gamma (IFNγ), interleukin (IL)-1β, IL-6, IL-10, transforming growth factor-beta (TGFβ) and cluster of differentiation (CD40) genes, as well as major histocompatibility complex (MHC)-II protein were evaluated. At 6 h post-stimulation, monobutyrin, calcium butyrate, and green tea extract treatments induced a significant downregulation of IFNγ, IL-6, or IL-1β gene transcription and MHC-II expression, while the IL-10 or TGFβ gene expression in these treatments as well as those receiving rosemary extract and tartaric acid was significantly upregulated, when compared to control, suggesting immunomodulatory properties of these ECs. Finally, pretreatment of macrophages with these selected 5 ECs for 24 h followed by C. perfringens infection showed that monobutyrin, green tea extract, rosemary extract, and calcium butyrate treatments can inhibit bacterial growth significantly at 12 and/or 24 h post-infection, when compared to the control. Collectively, our findings show that ECs possessing immunomodulatory and anti-C. perfringens properties can be selected using an in vitro avian macrophage cell-based model so that such ECs can further be tested in vivo for their disease prevention efficacy., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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4. IL-13 protects from Clostridioides difficile colitis.
- Author
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Donlan AN, Leslie JL, Simpson ME, Petri WA, Allen JE, and Petri WA
- Subjects
- Animals, Mice, Colitis immunology, Colitis prevention & control, Colitis microbiology, Mice, Inbred C57BL, Male, Clostridioides difficile immunology, Clostridium Infections prevention & control, Clostridium Infections immunology, Clostridium Infections microbiology, Disease Models, Animal, Interleukin-13 metabolism, Interleukin-13 immunology
- Abstract
Objectives: Clostridioides difficile infection (CDI) is the leading hospital-acquired infection in North America. We have previously discovered that antibiotic disruption of the gut microbiota decreases intestinal IL-33 and IL-25 and increases susceptibility to CDI. We further found that IL-33 promotes protection through type 2 Innate Lymphoid Cells (ILC2s), which produce IL-13. However, the contribution of IL-13 to disease has never been explored., Methods: We used a validated model of CDI in mice, in which we neutralized via blocking antibodies, or administered recombinant protein, IL-13 to assess the role of this cytokine during infection using weight and clinical scores. Fluorescent activated cell sorting (FACS) was used to characterize myeloid cell population changes in response to IL-13 manipulation., Results: We found that administration of IL-13 protected, and anti-IL-13 exacerbated CDI. Additionally, we observed alterations to the monocyte/macrophage cells following neutralization of IL-13 as early as day three post infection. We also observed elevated accumulation of myeloid cells by day four post-infection following IL-13 neutralization. Neutralization of the decoy receptor, IL-13Rα2, resulted in protection from disease, likely through increased available endogenous IL-13., Conclusions: Our data highlight the protective role of IL-13 in protecting from more severe CDI and the association of poor responses with a dysregulated monocyte-macrophage compartment. These results increase our understanding of type 2 immunity in CDI and may have implications for treating disease in patients., Competing Interests: Declaration of competing interest Dr. Petri has a conflict of interest in that he is a consultant for TechLab, Inc, which makes diagnostic tests for C. difficile infection. The other authors have no other conflicts of interest to disclose., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
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- 2024
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5. Supplementation with organic yeast-derived selenium provides immune protection against experimental necrotic enteritis in broiler chickens.
- Author
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Zhang M, Liu J, Yu Z, Chen Z, Yang J, Yin Y, and Xu S
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- Animals, Bacterial Toxins immunology, Necrosis, beta-Defensins metabolism, Jejunum drug effects, Jejunum immunology, Jejunum microbiology, Jejunum pathology, Spleen immunology, Yeasts, Nitric Oxide Synthase Type II metabolism, Interleukin-6 metabolism, Interleukin-8 metabolism, Interleukin-1beta metabolism, Antibodies, Bacterial blood, Chickens, Enteritis prevention & control, Enteritis veterinary, Enteritis immunology, Enteritis microbiology, Selenium pharmacology, Selenium administration & dosage, Poultry Diseases prevention & control, Poultry Diseases immunology, Dietary Supplements, Clostridium perfringens immunology, Clostridium Infections prevention & control, Clostridium Infections veterinary, Clostridium Infections immunology, Animal Feed, Cytokines metabolism
- Abstract
Necrotic enteritis (NE) is a potentially fatal poultry disease that causes enormous economic losses in the poultry industry worldwide. The study aimed to evaluate the effects of dietary organic yeast-derived selenium (Se) on immune protection against experimental necrotic enteritis (NE) in commercial broilers. Chickens were fed basal diets supplemented with different Se levels (0.25, 0.50, and 1.00 Se mg/kg). To induce NE, Clostridium perfringens (C. perfringens) was orally administered at 14 days of age post hatch. The results showed that birds fed 0.25 Se mg/kg exhibited significantly increased body weight gain compared with the non-supplemented/infected birds. There were no significant differences in gut lesions between the Se-supplemented groups and the non-supplemented group. The antibody levels against α-toxin and NetB toxin increased with the increase between 0.25 Se mg/kg and 0.50 Se mg/kg. In the jejunal scrapings and spleen, the Se-supplementation groups up-regulated the transcripts for pro-inflammatory cytokines IL-1β, IL-6, IL-8, iNOS, and LITAF and avian β-defensin 6, 8, and 13 (AvBD6, 8 and 13). In conclusion, supplementation with organic yeast-derived Se alleviates the negative consequences and provides beneficial protection against experimental NE., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Meiyu Zhang, Zehai Yu, Jian Liu, Zhiyuan Chen, Jiehua Yang, Yanbo Yin and Shouzhen Xu reports financial support was provided by The Industry Innovation Team of the Modern Agricultural Technology System of Shandong Province. Shouzhen Xu reports a relationship with The Industry Innovation Team of the Modern Agricultural Technology System of Shandong Province that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
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- 2024
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6. Effect of arginine supplementation on the growth performance, intestinal health, and immune responses of broilers during necrotic enteritis challenge.
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Fathima S, Hakeem WGA, Shanmugasundaram R, and Selvaraj RK
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- Animals, Eimeria physiology, Clostridium perfringens physiology, Male, Clostridium Infections veterinary, Clostridium Infections immunology, Arginine administration & dosage, Arginine pharmacology, Chickens growth & development, Chickens immunology, Poultry Diseases immunology, Dietary Supplements analysis, Enteritis veterinary, Enteritis immunology, Animal Feed analysis, Diet veterinary, Coccidiosis veterinary, Coccidiosis immunology, Random Allocation, Intestines drug effects
- Abstract
The objective of this study was to evaluate the effect of 25% arginine supplementation as a functional amino acid in partially alleviating the detrimental effects of necrotic enteritis (NE) on the growth performance, serum biochemistry, gut integrity, and the relative gene expression of tight junction proteins and inflammatory cytokines in broilers during NE. Three hundred and sixty 1-day-old chicks were randomly allocated to 4 treatments in a 2 × 2 factorial arrangement -basal diet and 125% arginine diet, with or without NE challenge. NE was induced by inoculating 1 × 10
4 Eimeria maxima sporulated oocysts on d 14 and 1 × 108 CFU/bird C. perfringens on d 19, 20, and 21. The NE challenge had a significant effect on the BWG (p < 0.05), FCR (p < 0.05), serum AST (p < 0.05), GLU (p < 0.05), and K+ (p < 0.05) levels, and intestinal permeability (p < 0.05) and jejunal lesion score (p < 0.05). A significant challenge × diet interaction effect was observed in the cecal tonsil CD8+ : CD4+ T-cell ratio on d 21 (p < 0.05) and 28 (p < 0.05) and spleen CD8+ : CD4+ T-cell ratio on d 21 (p < 0.05) and 35 (p < 0.05). Arginine supplementation significantly increased the CD8+ : CD4+ T-cell ratio in uninfected birds but decreased the CD8+ : CD4+ T-cell ratio in infected birds. On d 21, a significant interaction effect was observed on the relative expression of the iNOS gene (p < 0.05). Arginine supplementation significantly downregulated the expression of the iNOS gene in infected birds. A significant effect of the challenge (p < 0.05) was observed on the relative gene expression of the ZO-1 gene in the jejunum. NE challenge significantly downregulated the expression of the ZO-1 gene on d 21. In conclusion, arginine supplementation did not alleviate the depression in growth performance and disease severity during the NE challenge. However, arginine downregulated the expression of inflammatory cytokines and enzymes, preventing inflammatory injury to the tissues during NE. Hence, arginine might be supplemented with other alternatives to downregulate inflammatory response during NE in poultry., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2024
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7. Effect of 125% and 135% arginine on the growth performance, intestinal health, and immune responses of broilers during necrotic enteritis challenge.
- Author
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Fathima S, Al Hakeem WG, Shanmugasundaram R, Periyannan V, Varadhan R, and Selvaraj RK
- Subjects
- Animals, Eimeria physiology, Intestines drug effects, Clostridium Infections veterinary, Clostridium Infections immunology, Dose-Response Relationship, Drug, Male, Immunity, Innate drug effects, Chickens growth & development, Chickens immunology, Arginine administration & dosage, Arginine pharmacology, Poultry Diseases immunology, Poultry Diseases microbiology, Enteritis veterinary, Enteritis immunology, Animal Feed analysis, Diet veterinary, Dietary Supplements analysis, Clostridium perfringens physiology, Coccidiosis veterinary, Coccidiosis immunology, Random Allocation
- Abstract
The objective of this study was to evaluate the effects of 25% and 35% arginine supplementation in partially alleviating the effects of necrotic enteritis (NE) challenge on the production performance, intestinal integrity, and relative gene expression of tight junction proteins and inflammatory cytokines in broilers. Four hundred and eighty 1-day-old chicks were randomly allocated to the 4 treatments- Uninfected + Basal, NE + Basal, NE + Arg 125%, and NE + Arg 135%. NE was induced by inoculating 1 × 10
4 Eimeria maxima sporulated oocysts on d 14 and 1 × 108 CFU/bird C. perfringens on d 19, 20, and 21 of age by oral gavage. The NE challenge significantly decreased body weight gain (BWG) (p < 0.05) and increased the feed conversion ratio (FCR) (p < 0.05). On d 21, the NE challenge also increased the jejunal lesion score (p < 0.05) and relative gene expression of IL-10 and decreased the expression of the tight junction proteins occludin (p < 0.05) and claudin-4 (p < 0.05). The 125% arginine diet significantly increased intestinal permeability (p < 0.05) and the relative gene expression of iNOS (p < 0.05) and IFN-γ (p < 0.05) on d 21 and the bile anti-C. perfringens IgA concentration by 39.74% (p < 0.05) on d 28. The 135% arginine diet significantly increased the feed intake during d 0 - 28 (p < 0.05) and 0 to 35 (p < 0.05) and increased the FCR on d 0 to 35 (p < 0.05). The 135% and 125% arginine diet increased the spleen CD8+ : CD4+ T-cell ratio on d 28 (p < 0.05) and 35 (p < 0.05), respectively. The 135% arginine diet increased the CT CD8+ :CD4+ T-cell ratio on d 35 (p < 0.05). In conclusion, the 125% and 135% arginine diets did not reverse the effect of the NE challenge on the growth performance. However, the 125% arginine diet significantly increased the cellular and humoral immune response to the challenge. Hence, the 125% arginine diet could be used with other feed additives to improve the immune response of the broilers during the NE challenge., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2024
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8. Clostridium perfringens antigens and challenges for development of vaccines against necrotic enteritis in poultry.
- Author
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Waller SB, Galvão CC, Rodrigues RR, Aguirres CL, Quatrin PHDN, Alves MLF, Ferreira MRA, and Conceição FR
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- Animals, Antigens, Bacterial immunology, Vaccine Development methods, Vaccination veterinary, Vaccination methods, Necrosis veterinary, Clostridium perfringens immunology, Clostridium perfringens genetics, Enteritis prevention & control, Enteritis veterinary, Enteritis microbiology, Enteritis immunology, Bacterial Vaccines immunology, Bacterial Vaccines administration & dosage, Poultry Diseases prevention & control, Poultry Diseases microbiology, Clostridium Infections prevention & control, Clostridium Infections veterinary, Clostridium Infections immunology, Chickens
- Abstract
Introduction: Chickens with Necrotic Enteritis (NE), caused by Clostridium perfringens, exhibit acute and chronic symptoms that are difficult to diagnose, leading to significant economic losses. Vaccination is the best method for controlling and preventing NE. However, only two vaccines based on the CPA and NetB toxins have been commercialized, offering partial protection, highlighting the urgent need for more effective vaccines., Objective: This review aimed to identify promising antigens for NE vaccine formulation and discuss factors affecting their effectiveness., Methods: A systematic review using five scientific databases identified 30 eligible studies through the Rayyan tool, which were included for quality review., Results: We identified 25 promising antigens, including CPA, NetB, FBA, ZMP, CnaA, FimA, and FimB, categorized by their role in disease pathogenesis. This review discusses the biochemical, physiological, and genetic traits of recombinant antigens used in vaccine prototypes, their expression systems, and immunization potential in chickens challenged with virulent C. perfringens strains. Market supply challenges, immunogenic potential, vaccine platforms, adjuvants, and factors related to vaccination schedules-such as administration routes, dosing intervals, and age at immunization-are also addressed. Additionally, the study notes that vaccine formulations tested under mild challenges may not offer adequate field-level protection due to issues replicating aggressive conditions, strain virulence loss, and varied methodologies., Conclusions: An ideal NE vaccine should incorporate multiple antigens, molecular adjuvants, and delivery systems via in ovo and oral routes. The review underscores the challenges in developing and validating NE vaccines and the urgent need for a standardized protocol to replicate aggressive challenges accurately., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
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- 2024
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9. Development of two recombinant vaccines against Clostridioides difficile infection and immunogenicity in pregnant sows and neonatal piglets.
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Ramos CP, Siqueira WF, Viana LA, Cunha JLR, Fujiwara RT, Amarante VS, Souza TGV, and Silva ROS
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- Animals, Female, Swine, Rabbits, Pregnancy, Bacterial Proteins immunology, Bacterial Proteins genetics, Enterotoxins immunology, Enterotoxins genetics, Clostridium Infections prevention & control, Clostridium Infections veterinary, Clostridium Infections immunology, Bacterial Vaccines immunology, Bacterial Vaccines administration & dosage, Bacterial Vaccines genetics, Vaccines, Synthetic immunology, Vaccines, Synthetic administration & dosage, Clostridioides difficile immunology, Clostridioides difficile genetics, Animals, Newborn, Antibodies, Bacterial blood, Bacterial Toxins immunology, Bacterial Toxins genetics, Swine Diseases prevention & control, Swine Diseases immunology
- Abstract
Introduction: Clostridioides difficile is the main cause of antibiotic-associated diarrhea in humans and is a major enteropathogen in several animal species. In newborn piglets, colonic lesions caused by C. difficile A and B toxins (TcdA and TcdB, respectively) cause diarrhea and significant production losses., Objective: The present study aimed to develop two recombinant vaccines from immunogenic C-terminal fragments of TcdA and TcdB and evaluate the immune response in rabbits and in breeding sows. Two vaccines were produced: bivalent (rAB), consisting of recombinant fragments of TcdA and TcdB, and chimeric (rQAB), corresponding to the synthesis of the same fragments in a single protein. Groups of rabbits were inoculated with 10 or 50 μg of proteins adjuvanted with aluminum or 0.85 % sterile saline in a final volume of 1 mL/dose. Anti-TcdA and anti-TcdB IgG antibodies were detected in rabbits and sows immunized with both rAB and rQAB vaccines by ELISA. The vaccinated sows were inoculated intramuscularly with 20 μg/dose using a prime-boost approach., Results: Different antibody titers (p ≤ 0.05) were observed among the vaccinated groups of sows (rAB and rQAB) and control. Additionally, newborn piglets from vaccinated sows were also positive for anti-TcdA and anti-TcdB IgGs, in contrast to control piglets (p ≤ 0.05). Immunization of sows with the rQAB vaccine conferred higher anti-TcdA and anti-TcdB responses in piglets, suggesting the superiority of this compound over rAB., Conclusion: The synthesized recombinant proteins were capable of inducing antibody titers against C. difficile toxins A and B in sows, and were passively transferred to piglets through colostrum., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
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- 2024
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10. Production of highly soluble and immuno-reactive recombinant flagellin protein of Clostridium chauvoei.
- Author
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Prajapati A, Hemanth RA, Namrutha MR, Bindu S, Yogisharadhya R, Mohanty NN, Chanda MM, and Shivachandra SB
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- Animals, Rabbits, Guinea Pigs, Bacterial Vaccines immunology, Bacterial Vaccines genetics, Clostridium Infections veterinary, Clostridium Infections immunology, Clostridium Infections microbiology, Cattle, Antigens, Bacterial immunology, Antigens, Bacterial genetics, Cattle Diseases immunology, Cattle Diseases microbiology, Escherichia coli genetics, Escherichia coli metabolism, Enzyme-Linked Immunosorbent Assay, Cloning, Molecular, Flagellin immunology, Flagellin genetics, Clostridium chauvoei immunology, Clostridium chauvoei genetics, Recombinant Proteins immunology, Recombinant Proteins genetics, Antibodies, Bacterial blood, Antibodies, Bacterial immunology
- Abstract
Objective: Flagellin protein, an integral component of flagella, provides motility to several bacterial species and also acts as a candidate antigen in diagnostics and subunit vaccines. The bulk production of flagellin with retention of all conformational epitopes using recombinant protein technology is of paramount importance in the development of pathogen-specific immuno-assays and vaccines. We describe the production of highly soluble and immuno-reactive rFliA(C) protein of Clostridium chauvoei, a causative agent of blackleg or black quarter (BQ) affecting cattle and small ruminants worldwide. The bacterium is known to possess peritrichous flagella that provide motility and also act as a virulence factor with high protective antigenicity., Methods: Upon sequence and structural analysis, a partial fliA(C) gene from Clostridium chauvoei was cloned and the recombinant mature protein with N- and C- terminal truncation was over-expressed as a His-tagged fusion protein (∼25 kDa) in Escherichia coli. Subsequently, rFliA(C) protein was purified by single-step affinity chromatography and characterized for its immuno-reactivity in laboratory animals, Western blot, and indirect-ELISA format., Results: rFliA(C) was highly soluble and was purified in high quantity and quality. rFliA(C) elicited antigen-specific conformational polyclonal antibodies in rabbit and guinea pig models, as well as anti-Clostridium chauvoei-specific antibodies being specifically detected in BQ-vaccinated and convalescent sera of bovines in Western blot and in indirect-ELISA format. Further, no cross reactivity was noted with antibodies against major bovine diseases (e.g., foot-and-mouth disease, IBR, LSDV, hemorrhagic septicaemia, brucellosis, and leptospirosis)., Conclusion: The study indicated the production of conformational recombinant flagellin-rFliA(C)-antigen and its potential utility in development of diagnostics for detection of Clostridium chauvoei-specific antibodies in BQ-recovered and/or vaccinated animals., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
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- 2024
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11. Cell-Free Supernatant of Bacillus subtilis G2B9-Q Improves Intestinal Health and Modulates Immune Response to Promote Mouse Recovery in Clostridium perfringens Infection.
- Author
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Xu Z, Feng X, Song Z, Li X, Li K, Li M, Wang X, Liu B, and Sun C
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- Animals, Mice, Intestines microbiology, Intestines immunology, Swine, Diarrhea microbiology, Diarrhea immunology, Feces microbiology, Disease Models, Animal, Clostridium Infections immunology, Clostridium Infections microbiology, Bacillus subtilis genetics, Clostridium perfringens immunology, Probiotics administration & dosage, Gastrointestinal Microbiome drug effects
- Abstract
Clostridium perfringens is one of the critical causative agents causing diarrhea in piglets, with significant economic losses to the pig industry. Under normal gut microbiota homeostasis and well-managed barns, diarrhea caused by C. perfringens could be controlled. Some reports show that probiotics, such as Bacillus subtilis, are beneficial in preventing necrotic enteritis (NE) in chickens, but few reports on piglets. Clostridium perfringens was found in the piglets' diarrhea with intestinal microbiota dysbiosis in our survey. Bacillus subtilis G2B9-Q, which was isolated from the feces of healthy pigs, was found to have anti-Clostridium activity after screening. Clostridium perfringens was used to challenge mice by intraperitoneal injection for modeling to evaluate the anti-infective activity of cell-free supernatant (CFS) of B. subtilis G2B9-Q and different concentrations of B. subtilis G2B9-Q by oral administration. The results showed that G2B9-Q can mitigate intestinal lesions caused by C. perfringens infection, reduce inflammatory reactions, and modulate intestinal microbiota. The CFS of G2B9-Q can alleviate the pathological damage of intestinal tissues caused by C. perfringens infection, reduce the concentration of TNF-α and IL-10 in the sera of mice, as well as the relative expression levels of alpha toxin (CPA), perfringolysin O (PFO) toxin, IL-10, IL-22, and TNF-α in the jejunum and colon tissues, and alleviate the changes in gut microbiota structure caused by C. perfringens infection, which showed better therapeutic effects and indicated that the metabolites of G2B9-Q are essential mediators for their beneficial effects. Therefore, the CFS of G2B9-Q could potentially replace antibiotics in treating C. perfringens infection., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
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- 2024
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12. Absent in Melanoma 2 Mediates Inflammasome Signaling Activation against Clostridium perfringens Infection.
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Ma Z, Lou Y, Wang N, Zhao Y, Zhang S, Zhang M, Li J, Xu Q, He A, and Yu S
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- Animals, Mice, Mice, Knockout, Immunity, Innate, Mice, Inbred C57BL, Gas Gangrene immunology, Gas Gangrene microbiology, Disease Models, Animal, Clostridium Infections immunology, Clostridium Infections microbiology, Clostridium Infections metabolism, Humans, Inflammasomes metabolism, Inflammasomes immunology, Clostridium perfringens immunology, Clostridium perfringens pathogenicity, Signal Transduction, DNA-Binding Proteins metabolism, DNA-Binding Proteins genetics
- Abstract
Absent in melanoma 2 (AIM2), a key component of the IFI20X/IFI16 (PYHIN) protein family, is characterized as a DNA sensor to detect cytosolic bacteria and DNA viruses. However, little is known about its immunological role during pathogenic Clostridium perfringens ( C . perfringens ) infection, an extracellular bacterial pathogen. In a pathogenic C . perfringens gas gangrene model, Aim2
-/- mice are more susceptible to pathogenic C . perfringens soft tissue infection, revealing the importance of AIM2 in host protection. Notably, Aim2 deficiency leads to a defect in bacterial killing and clearance. Our in vivo and in vitro findings further establish that inflammasome signaling is impaired in the absence of Aim2 in response to pathogenic C . perfringens . Mechanistically, inflammasome signaling downstream of active AIM2 promotes pathogen control. Importantly, pathogenic C . perfringens -derived genomic DNA triggers inflammasome signaling activation in an AIM2-dependent manner. Thus, these observations uncover a central role for AIM2 in host defense and triggering innate immunity to combat pathogenic C . perfringens infections.- Published
- 2024
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13. The impact of existing total anti-toxin B IgG immunity in outcomes of recurrent Clostridioides difficile infection.
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Rigo I, Young MK, Abhyankar MM, Xu F, Ramakrishnan G, Naz F, Madden GR, and Petri WA
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- Humans, Male, Middle Aged, Female, Aged, Bacterial Proteins immunology, Prospective Studies, Antibodies, Neutralizing immunology, Antibodies, Neutralizing blood, Adult, Aged, 80 and over, Clostridium Infections immunology, Clostridium Infections prevention & control, Immunoglobulin G blood, Immunoglobulin G immunology, Bacterial Toxins immunology, Clostridioides difficile immunology, Recurrence, Antibodies, Bacterial blood, Antibodies, Bacterial immunology
- Abstract
Late anti-toxin-B humoral immunity acquired after treatment is important for preventing recurrent Clostridioides difficile infection. We prospectively-measured anti-toxin-B IgG and neutralization titers at diagnosis as potential early predictors of recurrence. High anti-toxin-B-IgG/neutralizing antibodies were associated with short-lasting protection within 6-weeks, however, no difference in recurrence risk was observed by 90-days post-infection., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: William A. Petri Jr is a consultant for TechLab, a company that makes diagnostics for C. difficile. The other authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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14. Systemic neutrophil degranulation and emergency granulopoiesis in patients with Clostridioides difficile infection.
- Author
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Ramakrishnan G, Young MK, Nayak U, Rigo I, Marrs AS, Gilchrist CA, Behm BW, Madden GR, and Petri WA Jr
- Subjects
- Humans, Male, Female, Middle Aged, Aged, Biomarkers blood, Adult, Flow Cytometry, Neutrophil Activation, Aged, 80 and over, Cytokines blood, Lipocalin-2 blood, Neutrophils immunology, Cell Degranulation, Clostridium Infections immunology, Clostridium Infections blood, Clostridium Infections microbiology, Clostridioides difficile
- Abstract
Objectives: Clostridioides difficile infection (CDI) is characterized by neutrophilia in blood, with a high leukocyte count accompanying severe infection. In this study, we characterized peripheral blood neutrophil activation and maturity in CDI by (i) developing a method to phenotype stored neutrophils for disease-related developmental alterations and (ii) assessing neutrophil-associated biomarkers., Methods: We stored fixed leukocytes from blood collected within 24 h of diagnosis from a cohort of hospitalized patients with acute CDI. Additional study cohorts included recurrent CDI patients at time of and two months after FMT therapy and a control healthy cohort. We assessed levels of neutrophil surface markers CD66b, CD11b, CD16 and CD10 by flow cytometry. Plasma neutrophil elastase and lipocalin-2 were measured using ELISA, while G-CSF, GM-CSF and cytokines were measured using O-link Proteomic technology., Results: CD66b
+ neutrophil abundance assessed by flow cytometry correlated well with complete blood counts, establishing that neutrophils in stored blood are sufficiently well-preserved for phenotyping by flow cytometry. Neutrophil abundance was significantly increased in CDI patients compared to healthy controls. Emergency granulopoiesis in acute CDI patients was evidenced by lower neutrophil surface expression of CD10, CD11b and CD16. CD10+ staining of neutrophils started to recover within 3-7 days of CDI treatment. Neutrophil activation and degranulation were higher in acute CDI as assessed by plasma neutrophil elastase and lipocalin-2. Biomarker levels in immunocompetent subjects were associated with recurrence and fatal outcomes., Conclusions: Neutrophil activation and emergency granulopoiesis characterize the early immune response in acute CDI, with plasma degranulation biomarkers predictive of disease severity., Competing Interests: Declaration of competing interest WAP has a conflict of interest in that he is a consultant for TechLab, Inc, which makes diagnostic tests for C. difficile infection. The other authors have no conflicts of interest to disclose., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)- Published
- 2024
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15. Treatment of chickens with lactobacilli prior to challenge with Clostridium perfringens modifies innate responses and gut morphology.
- Author
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Alizadeh M, Shojadoost B, Fletcher C, Wang A, Abdelaziz K, and Sharif S
- Subjects
- Animals, Male, Intestines microbiology, Enteritis veterinary, Enteritis microbiology, Enteritis immunology, Chickens immunology, Chickens microbiology, Clostridium perfringens physiology, Clostridium Infections veterinary, Clostridium Infections immunology, Clostridium Infections therapy, Clostridium Infections microbiology, Poultry Diseases microbiology, Poultry Diseases immunology, Probiotics administration & dosage, Probiotics pharmacology, Immunity, Innate, Lactobacillus
- Abstract
Necrotic enteritis caused by Clostridium perfringens (CP), is a common enteric disease of poultry that has been previously controlled by in-feed antibiotics. However, due to the rapid emergence of antimicrobial resistance, alternatives to antibiotics such as probiotics have received considerable attention because of their immunomodulatory and intestinal health benefits. The present study investigated the effects of probiotic lactobacilli on gut histomorphology and intestinal innate responses in chickens. Day-old male broiler chickens were treated with 1 × 10
7 or 1 × 108 colony-forming units (CFU) of a lactobacilli cocktail on days 1, 7, 14, and 20 post-hatch, while control groups were not treated with lactobacilli. On day 21, birds in all groups (except the negative control) were challenged with 3 × 108 CFU of CP for 3 days. Intestinal tissue samples were collected before and after the CP challenge to assess gene expression and for histomorphological analysis. Lactobacilli treatment at a dose of 1 × 108 CFU conferred partial protection against NE by lowering lesion scores, increasing villus height in the ileum and reducing crypt depth in the jejunum. In addition, 1 × 108 CFU of lactobacilli enhanced the expression of Toll-like receptor (TLR) 2, interferon-gamma (IFN-γ), interleukin (IL)-10, IL-12, and IL-13 in both the jejunum and ileum at different timepoints and subsequently decreased the expression of transforming growth factor beta (TGF-β) and IL-1β post-CP challenge. In conclusion, the results indicate that treatment with lactobacilli mitigated NE in a dose-dependent manner via improvement of intestinal morphology and modulation of innate immune response in chickens., Competing Interests: Declaration of competing interest Authors declare no financial conflict of interest., (Copyright © 2024. Published by Elsevier Ltd.)- Published
- 2024
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16. Clostridioides difficile toxin B subverts germinal center and antibody recall responses by stimulating a drug-treatable CXCR4-dependent mechanism.
- Author
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Norman KM, Lang GA, Shadid TM, Honold ST, Reel JM, Cox MA, Ballard JD, and Lang ML
- Subjects
- Animals, Mice, Mice, Inbred C57BL, B-Lymphocytes immunology, B-Lymphocytes metabolism, Chemokine CXCL12 metabolism, Clostridium Infections immunology, Clostridium Infections microbiology, Humans, Immunoglobulin G immunology, Immunoglobulin G metabolism, Immunologic Memory, Female, Antibody Formation immunology, Receptors, CXCR4 metabolism, Receptors, CXCR4 immunology, Germinal Center immunology, Bacterial Proteins metabolism, Bacterial Proteins immunology, Bacterial Toxins immunology, Bacterial Toxins metabolism, Clostridioides difficile immunology, Clostridioides difficile pathogenicity
- Abstract
Recurrent Clostridioides difficile infection (CDI) results in significant morbidity and mortality. We previously established that CDI in mice does not protect against reinfection and is associated with poor pathogen-specific B cell memory (Bmem), recapitulating our observations with human Bmem. Here, we demonstrate that the secreted toxin TcdB2 is responsible for subversion of Bmem responses. TcdB2 from an endemic C. difficile strain delayed immunoglobulin G (IgG) class switch following vaccination, attenuated IgG recall to a vaccine booster, and prevented germinal center formation. The mechanism of TcdB2 action included increased B cell CXCR4 expression and responsiveness to its ligand CXCL12, accounting for altered cell migration and a failure of germinal center-dependent Bmem. These results were reproduced in a C. difficile infection model, and a US Food and Drug Administration (FDA)-approved CXCR4-blocking drug rescued germinal center formation. We therefore provide mechanistic insights into C. difficile-associated pathogenesis and illuminate a target for clinical intervention to limit recurrent disease., Competing Interests: Declaration of interests K.M.N., M.L.L., and J.D.B. are listed as inventors on provisional US patent OKLA.P0023US.P1 filed on 09/13/23, which is related to the work described here., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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17. Deficiency of IL-22-binding protein enhances the ability of the gut microbiota to protect against enteric pathogens.
- Author
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Fachi JL, Di Luccia B, Gilfillan S, Chang HW, Song C, Cheng J, Cella M, Vinolo MA, Gordon JI, and Colonna M
- Subjects
- Animals, Mice, Receptors, Interleukin metabolism, Receptors, Interleukin genetics, Interleukins metabolism, Mice, Inbred C57BL, Clostridium Infections immunology, Clostridium Infections microbiology, Clostridium Infections prevention & control, Gastrointestinal Microbiome, Citrobacter rodentium, Mice, Knockout, Clostridioides difficile, Interleukin-22, Enterobacteriaceae Infections immunology, Enterobacteriaceae Infections microbiology, Enterobacteriaceae Infections prevention & control
- Abstract
Interleukin 22 (IL-22) promotes intestinal barrier integrity, stimulating epithelial cells to enact defense mechanisms against enteric infections, including the production of antimicrobial peptides. IL-22 binding protein (IL-22BP) is a soluble decoy encoded by the Il22ra2 gene that decreases IL-22 bioavailability, attenuating IL-22 signaling. The impact of IL-22BP on gut microbiota composition and functioning is poorly understood. We found that Il22ra2
-/- mice are better protected against Clostridioides difficile and Citrobacter rodentium infections. This protection relied on IL-22-induced antimicrobial mechanisms before the infection occurred, rather than during the infection itself. Indeed, the gut microbiota of Il22ra2-/- mice mitigated infection of wild-type (WT) mice when transferred via cohousing or by cecal microbiota transplantation. Indicator species analysis of WT and Il22ra2-/- mice with and without cohousing disclosed that IL22BP deficiency yields a gut bacterial composition distinct from that of WT mice. Manipulation of dietary fiber content, measurements of intestinal short-chain fatty acids and oral treatment with acetate disclosed that resistance to C. difficile infection is related to increased production of acetate by Il22ra2-/- -associated microbiota. Together, these findings suggest that IL-22BP represents a potential therapeutic target for those at risk for or with already manifest infection with this and perhaps other enteropathogens., Competing Interests: Competing interests statement:The authors declare no competing interest.- Published
- 2024
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18. Dysregulated Immunity to Clostridioides difficile in IBD Patients Without a History of Recognized Infection.
- Author
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Cook L, Wong MQ, Rees WD, Schick A, Lisko DJ, Lunken GR, Wang X, Peters H, Oliveira L, Lau T, Mah R, Bressler B, Levings MK, and Steiner TS
- Subjects
- Humans, Male, Female, Adult, Case-Control Studies, Middle Aged, Bacterial Toxins immunology, Colitis, Ulcerative immunology, Colitis, Ulcerative microbiology, Th17 Cells immunology, Inflammatory Bowel Diseases immunology, Inflammatory Bowel Diseases microbiology, RNA, Ribosomal, 16S genetics, RNA, Ribosomal, 16S analysis, Crohn Disease immunology, Crohn Disease microbiology, Integrin beta Chains metabolism, Young Adult, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium Infections microbiology, Feces microbiology, Gastrointestinal Microbiome immunology, CD4-Positive T-Lymphocytes immunology, Bacterial Proteins
- Abstract
Background & Aims: Clostridioides difficile is a toxin-secreting bacteria that is an urgent antimicrobial resistance threat, with approximately 25% of patients developing recurrent infections. Inflammatory bowel disease (IBD) patients are at increased risk of severe, recurrent C. difficile infection., Methods: To investigate a role for C. difficile infection in IBD pathogenesis, we collected peripheral blood and stool from 20 each of ulcerative colitis patients, Crohn's disease patients, and healthy control subjects. We used a flow cytometric activation induced marker assay to quantify C. difficile toxin-specific CD4+ T cells and 16S ribosomal RNA sequencing to study microbiome diversity., Results: We found IBD patients had significantly increased levels of C. difficile toxin B-specific CD4+ T cells, but not immunoglobulin G or immunoglobulin A, compared with healthy control subjects. Within antigen-specific CD4+ T cells, T helper type 17 cells and cells expressing the gut homing receptor integrin β7 were reduced compared with healthy control subjects, similar to our previous study of non-IBD patients with recurrent C. difficile infection. Stool microbiome analysis revealed that gut homing, toxin-specific CD4+ T cells negatively associated with microbial diversity and, along with T helper type 17 cells, positively associated with bacteria enriched in healthy control subjects., Conclusions: These data suggest that IBD patients, potentially due to underlying intestinal dysbiosis, experience undiagnosed C. difficile infections that result in impaired toxin-specific immunity. This may contribute to the development of inflammatory T cell responses toward commensal bacteria and provide a rationale for C. difficile testing in IBD patients., (© 2023 Crohn’s & Colitis Foundation. Published by Oxford University Press on behalf of Crohn’s & Colitis Foundation.)
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- 2024
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19. Lactobacilli-derived adjuvants combined with immunoinformatics-driven multi-epitope antigens based approach protects against Clostridium perfringens in a mouse model.
- Author
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Guo Z, Ren H, Chang Q, Liu R, Zhou X, Xue K, Sun T, Luo J, Wang F, and Ge J
- Subjects
- Animals, Mice, Computational Biology, Antigens, Bacterial immunology, Female, Mice, Inbred BALB C, Immunoinformatics, Clostridium perfringens immunology, Lactobacillus immunology, Adjuvants, Immunologic, Epitopes immunology, Bacterial Vaccines immunology, Clostridium Infections prevention & control, Clostridium Infections immunology, Disease Models, Animal
- Abstract
Clostridium perfringens is ubiquitously distributed and capable of secreting toxins, posing a significant threat to animal health. Infections caused by Clostridium perfringens, such as Necrotic Enteritis (NE), result in substantial economic losses to the livestock industry annually. However, there is no effective commercial vaccine available. Hence, we set out to propose an effective approach for multi-epitope subunit vaccine construction utilizing biomolecules. We utilized immunoinformatics to design a novel multi-epitope antigen against C. perfringens (CPMEA). Furthermore, we innovated novel bacterium-like particles (BLPs) through thermal acid treatment of various Lactobacillus strains and selected BLP23017 among them. Then, we detailed the structure of CPMEA and BLPs and utilized them to prepare a multi-epitope vaccine. Here, we showed that our vaccine provided full protection against C. perfringens infection after a single dose in a mouse model. Additionally, BLP23017 notably augmented the secretion of secretory immunoglobulin A (sIgA) and enhanced antibody production. We conclude that our vaccine possess safety and high efficacy, making it an excellent candidate for preventing C. perfringens infection. Moreover, we demonstrate our approach to vaccine construction and the preparation of BLP23017 with distinct advantages may contribute to the prevention of a wider array of diseases and the novel vaccine development., Competing Interests: Declaration of competing interest The authors of this paper declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
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20. Immunization of turkeys with Clostridium septicum alpha toxin-based recombinant subunit proteins can confer protection against experimental Clostridial dermatitis.
- Author
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John FA, Criollo V, Gaghan C, Armwood A, Holmes J, Thachil AJ, Crespo R, and Kulkarni RR
- Subjects
- Animals, Bacterial Vaccines immunology, Bacterial Vaccines administration & dosage, Immunization, Turkeys immunology, Clostridium septicum immunology, Clostridium Infections prevention & control, Clostridium Infections immunology, Clostridium Infections veterinary, Poultry Diseases prevention & control, Poultry Diseases immunology, Poultry Diseases microbiology, Bacterial Toxins immunology, Recombinant Proteins immunology, Recombinant Proteins administration & dosage, Dermatitis prevention & control, Dermatitis immunology, Dermatitis veterinary
- Abstract
Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ntATX-D1 and ntATX-D2, were identified, cloned, and expressed in Escherichia coli as recombinant subunit proteins to investigate their use as potential vaccine candidates. Experimental groups consisted of a Negative control (NCx) that did not receive C. septicum challenge, while the adjuvant-only Positive control (PCx), ntATX-D1 immunization (D1) and ntATX-D2 immunization (D2) groups received C. septicum challenge. Turkeys were immunized subcutaneously with 100 μg of protein at 7, 8 and 9 weeks of age along with an oil-in-water nano-emulsion adjuvant, followed by C. septicum challenge at 11 weeks of age. Results showed that while 46.2% of birds in the PCx group died post-challenge, the rate of mortality in D1- or D2-immunization groups was 13.3%. The gross and histopathological lesions in the skin, muscle and spleen showed that the disease severity was highest in PCx group, while the D2-immunized birds had significantly lower lesion scores when compared to PCx. Gene expression analysis revealed that PCx birds had significantly higher expression of pro-inflammatory cytokine genes in the skin, muscle and spleen than the NCx group, while the D2 group had significantly lower expression of these genes compared to PCx. Peripheral blood cellular analysis showed increased frequencies of activated CD4+ and/or CD8+ cells in the D1 and D2-immunized groups. Additionally, the immunized turkeys developed antigen-specific serum IgY antibodies. Collectively, these findings indicate that ntATX proteins, specifically the ntATX-D2 can be a promising vaccine candidate for protecting turkeys against CD and that the protection mechanisms may include downregulation of C. septicum-induced inflammation and increased CD4+ and CD8+ cellular activation., Competing Interests: US Provisional Patent Nos. 63/609,022, 63/608,491, 63/597,112 titled "Vaccine Composition Against Clostridial Dermatitis". This does not alter our adherence to PLOS ONE policies on sharing data and materials.The sequences of ntATX-D1 and ntATX-D2 vaccine antigens reported in this work are also deposited in the GenBank (https://www.ncbi.nlm.nih.gov/nuccore/PP003322 and https://www.ncbi.nlm.nih.gov/nuccore/PP003323)., (Copyright: © 2024 John et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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21. An in vitro assay for toxicity testing of Clostridium perfringens type C β-toxin.
- Author
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Hoonakker M, Zariri A, de Brouwer L, David D, Borgman A, and Sloots A
- Subjects
- Animals, Humans, Vero Cells, Chlorocebus aethiops, Toxicity Tests methods, Clostridium Infections veterinary, Clostridium Infections immunology, Clostridium Infections diagnosis, THP-1 Cells, Mice, Cell Survival drug effects, Cell Line, Bacterial Vaccines immunology, Animal Testing Alternatives methods, Clostridium perfringens immunology, Bacterial Toxins immunology, Bacterial Toxins toxicity
- Abstract
Introduction: Veterinary vaccines against Clostridium perfringens type C need to be tested for absence of toxicity, as mandated by pharmacopoeias worldwide. This toxicity testing is required at multiple manufacturing steps and relies on outdated mouse tests that involve severe animal suffering. Clostridium perfringens type C produces several toxins of which the β-toxin is the primary component responsible for causing disease. Here, we describe the successful development of a new cell-based in vitro assay that can address the specific toxicity of the β-toxin., Methods: Development of the cell-based assay followed the principle of in vitro testing developed for Cl. septicum vaccines, which is based on Vero cells. We screened four cell lines and selected the THP-1 cell line, which was shown to be the most specific and sensitive for β-toxin activity, in combination with a commercially available method to determine cell viability (MTS assay) as a readout., Results: The current animal test is estimated to detect 100 - 1000-fold dilutions of the Cl. perfringens type C non-inactivated antigen. When tested with an active Cl. perfringens type C antigen preparation, derived from a commercial vaccine manufacturing process, our THP-1 cell-based assay was able to detect toxin activity from undiluted to over 10000-fold dilution, showing a linear range between approximately 1000- and 10000-fold dilutions. Assay specificity for the β-toxin was confirmed with neutralizing antibodies and lack of reaction to Cl. perfringens culture medium. In addition, assay parameters demonstrated good repeatability., Conclusions: Here, we have shown proof of concept for a THP-1 cell-based assay for toxicity testing of veterinary Cl. perfringens type C vaccines that is suitable for all vaccine production steps. This result represents a significant step towards the replacement of animal-based toxicity testing of this veterinary clostridial antigen. As a next step, assessment of the assay's sensitivity and repeatability and validation of the method will have to be performed in a commercial manufacturing context in order to formally implement the assay in vaccine quality control., Competing Interests: All authors were employed by company Intravacc B.V., (Copyright © 2024 Hoonakker, Zariri, de Brouwer, David, Borgman and Sloots.)
- Published
- 2024
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22. Fecal microbiota transplantation stimulates type 2 and tolerogenic immune responses in a mouse model.
- Author
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Moreau GB, Naz F, and Petri WA Jr
- Subjects
- Animals, Mice, Gastrointestinal Microbiome, Dysbiosis therapy, Clostridioides difficile immunology, Immune Tolerance, Mice, Inbred C57BL, Fecal Microbiota Transplantation methods, Disease Models, Animal, Clostridium Infections therapy, Clostridium Infections immunology, Clostridium Infections microbiology
- Abstract
Objectives: Clostridioides difficile infection (CDI) is the leading hospital-acquired infection in North America. While previous work on fecal microbiota transplantation (FMT), a highly effective treatment for CDI, has focused on colonization resistance mounted against C. difficile by FMT-delivered commensals, the effects of FMT on host gene expression are relatively unexplored. This study aims to identify transcriptional changes associated with FMT, particularly changes associated with protective immune responses., Methods: Gene expression was assessed on day 2 and day 7 after FMT in mice after antibiotic-induced dysbiosis. Flow cytometry was also performed on colon and mesenteric lymph nodes at day 7 to investigate changes in immune cell populations., Results: FMT administration after antibiotic-induced dysbiosis successfully restored microbial alpha diversity to levels of donor mice by day 7 post-FMT. Bulk RNA sequencing of cecal tissue at day 2 identified immune genes, including both pro-inflammatory and Type 2 immune pathways as upregulated after FMT. RNA sequencing was repeated on day 7 post-FMT, and expression of these immune genes was decreased along with upregulation of genes associated with restoration of intestinal homeostasis. Immunoprofiling on day 7 identified increased colonic CD45
+ immune cells that exhibited dampened Type 1 and heightened regulatory and Type 2 responses. These include an increased abundance of eosinophils, alternatively activated macrophages, Th2, and T regulatory cell populations., Conclusion: These results highlight the impact of FMT on host gene expression, providing evidence that FMT restores intestinal homeostasis after antibiotic treatment and facilitates tolerogenic and Type 2 immune responses., Competing Interests: Declaration of competing interest Dr. Petri has a conflict of interest in that I am a consultant for TechLab, Inc., which makes diagnostic tests for C. difficile infection. The other authors have no other conflicts of interest to disclose., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)- Published
- 2024
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23. Clostridioides difficile ferrosome organelles combat nutritional immunity.
- Author
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Pi H, Sun R, McBride JR, Kruse ARS, Gibson-Corley KN, Krystofiak ES, Nicholson MR, Spraggins JM, Zhou Q, and Skaar EP
- Subjects
- Animals, Mice, Homeostasis, Bacterial Proteins metabolism, Cell Membrane metabolism, Cryoelectron Microscopy, Electron Microscope Tomography, Disease Models, Animal, Leukocyte L1 Antigen Complex metabolism, Microbial Viability, Inflammation metabolism, Inflammation microbiology, Intestines metabolism, Intestines microbiology, Clostridioides difficile growth & development, Clostridioides difficile immunology, Clostridioides difficile metabolism, Clostridium Infections immunology, Clostridium Infections metabolism, Clostridium Infections microbiology, Iron metabolism, Organelles metabolism, Ferric Compounds metabolism, Host Microbial Interactions
- Abstract
Iron is indispensable for almost all forms of life but toxic at elevated levels
1-4 . To survive within their hosts, bacterial pathogens have evolved iron uptake, storage and detoxification strategies to maintain iron homeostasis1,5,6 . Recent studies showed that three Gram-negative environmental anaerobes produce iron-containing ferrosome granules7,8 . However, it remains unclear whether ferrosomes are generated exclusively by Gram-negative bacteria. The Gram-positive bacterium Clostridioides difficile is the leading cause of nosocomial and antibiotic-associated infections in the USA9 . Here we report that C. difficile undergoes an intracellular iron biomineralization process and stores iron in membrane-bound ferrosome organelles containing non-crystalline iron phosphate biominerals. We found that a membrane protein (FezA) and a P1B6 -ATPase transporter (FezB), repressed by both iron and the ferric uptake regulator Fur, are required for ferrosome formation and play an important role in iron homeostasis during transition from iron deficiency to excess. Additionally, ferrosomes are often localized adjacent to cellular membranes as shown by cryo-electron tomography. Furthermore, using two mouse models of C. difficile infection, we demonstrated that the ferrosome system is activated in the inflamed gut to combat calprotectin-mediated iron sequestration and is important for bacterial colonization and survival during C. difficile infection., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2023
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24. Stool Interleukin-1β Differentiates Clostridioides difficile Infection (CDI) From Asymptomatic Carriage and Non-CDI Diarrhea.
- Author
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Villafuerte Gálvez JA, Pollock NR, Alonso CD, Chen X, Xu H, Wang L, White N, Banz A, Miller M, Daugherty K, Gonzalez-Luna AJ, Barrett C, Sprague R, Garey KW, and Kelly CP
- Subjects
- Humans, Antitoxins, Bacterial Toxins, Enterotoxins, Immunoglobulin A, Immunoglobulin G, Clostridioides difficile, Clostridium Infections complications, Clostridium Infections diagnosis, Clostridium Infections immunology, Diarrhea etiology, Feces chemistry, Interleukin-1beta
- Abstract
Background: Despite advances in the understanding and diagnosis of Clostridioides difficile infection (CDI), clinical distinction within the colonization-infection continuum remains an unmet need., Methods: By measuring stool cytokines and antitoxin antibodies in well-characterized cohorts of CDI (diarrhea, nucleic acid amplification test [NAAT] positive), non-CDI diarrhea (NCD; diarrhea, NAAT negative), asymptomatic carriers (ASC; no diarrhea, NAAT positive) and hospital controls (CON; no diarrhea, NAAT negative), we aim to discover novel biological markers to distinguish between these cohorts. We also explore the relationship of these stool cytokines and antitoxin antibody with stool toxin concentrations and disease severity., Results: Stool interleukin (IL) 1β, stool immunoglobulin A (IgA), and immunoglobulin G (IgG) anti-toxin A had higher (P < .0001) concentrations in CDI (n = 120) vs ASC (n = 43), whereas toxins A, B, and fecal calprotectin did not. Areas under the receiver operating characteristic curve (ROC-AUCs) for IL-1β, IgA, and IgG anti-toxin A were 0.88, 0.83, and 0.83, respectively. A multipredictor model including IL-1β and IgA anti-toxin A achieved an ROC-AUC of 0.93. Stool IL-1β concentrations were higher in CDI compared to NCD (n = 75) (P < .0001) and NCD + ASC+ CON (CON, n = 75) (P < .0001), with ROC-AUCs of 0.83 and 0.86, respectively. Stool IL-1β had positive correlations with toxins A (ρA = +0.55) and B (ρB = +0.49) in CDI (P < .0001) but not in ASC (P > .05)., Conclusions: Stool concentrations of the inflammasome pathway, proinflammatory cytokine IL-1β, can accurately differentiate CDI from asymptomatic carriage and NCD, making it a promising biomarker for CDI diagnosis. Significant positive correlations exist between stool toxins and stool IL-1β in CDI but not in asymptomatic carriers., Competing Interests: Potential conflicts of interest. C. D. A. has received grant support from Merck (investigator-initiated award, paid to institution); consulting fees for advisory board from Cidara Therapeutics, Merck, and Prime Meridian Group (on behalf of AiCuris); and honoraria for presentations from the American Society of Healthcare Pharmacists. A. B. reports being an employee of bioMérieux. M. M. reports being an employee of bioMérieux and a holder of patent (Susceptibility to C. difficile infection and Fecal Immunoglobulins; bioMérieux). C. P. K. has acted as a paid consultant to Artugen, Facile Therapeutics, Ferring, First Light Biosciences, Finch, Janssen (J&J), Milky Way Life Sciences, Pfizer, Seres, Summit, RVAC Medicines, and Vedanta; has received grant support from Milky Way Life Sciences; reports participation on a data and safety monitoring board or advisory board from Finch Therapeutics (payments to self); has served as Secretary (unpaid) for the Society for the Study of Celiac Disease; and holds stock or stock options from First Light. K. W. G. has received grant support paid to the University of Houston from Acurx, Summit, Paratek Pharmaceuticals, Tetraphase Pharmaceuticals, and Seres Health, and reports consulting fees from Acurx and Summit Pharmaceuticals. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2022. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2023
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25. Use of a Clostridioides difficile Murine Immunization and Challenge Model to Evaluate Single and Combination Vaccine Adjuvants Consisting of Alum and NKT Cell-Activating Ligands.
- Author
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Lang GA, Norman K, Amadou Amani S, Shadid TM, Ballard JD, and Lang ML
- Subjects
- Animals, Bacterial Vaccines administration & dosage, Biomarkers, Disease Models, Animal, Dose-Response Relationship, Immunologic, Female, Immunization, Immunophenotyping, Ligands, Lymphocyte Activation immunology, Mice, Natural Killer T-Cells metabolism, Adjuvants, Vaccine administration & dosage, Adjuvants, Vaccine chemistry, Alum Compounds, Bacterial Vaccines immunology, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium Infections prevention & control, Natural Killer T-Cells immunology
- Abstract
Adjuvant combinations may enhance or broaden the expression of immune responses to vaccine antigens. Information on whether established Alum type adjuvants can be combined with experimental CD1d ligand adjuvants is currently lacking. In this study, we used a murine Clostridioides difficile immunization and challenge model to evaluate Alum (Alhydrogel™), α-galactosylceramide (α-GC), and one of its analogs 7DW8-5 singly and in combination as vaccine adjuvants. We observed that the Alum/α-GC combination caused modest enhancement of vaccine antigen-specific IgG1 and IgG2b responses, and a broadening to include IgG2c that did not significantly impact overall protection. Similar observations were made using the Alum/7DW8-5 combination. Examination of the impact of adjuvants on NKT cells revealed expansion of invariant NKT (iNKT) cells with modest expansion of their iNKTfh subset and little effect on diverse NKT (dNKT) cells. Side effects of the adjuvants was determined and revealed transient hepatotoxicity when Alum/α-GC was used in combination but not singly. In summary these results showed that the Alum/α-GC or the Alum/7DW8-5 combination could exert distinct effects on the NKT cell compartment and on isotype switch to produce Th1-driven IgG subclasses in addition to Alum/Th2-driven subclasses. While Alum alone was efficacious in stimulating IgG-mediated protection, and α-GC offered no apparent additional benefit in the C. difficile challenge model, the work herein reveals immune response features that could be optimized and harnessed in other vaccine contexts., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lang, Norman, Amadou Amani, Shadid, Ballard and Lang.)
- Published
- 2022
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26. Effective definition of low humoral response to Clostridioides difficile infection.
- Author
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Ramos-Martínez A, Serrano-Martínez F, Pintos I, Valencia-Alijo Á, Gutiérrez-Rojas Á, Cítores MJ, Ortiz-Balbuena J, Royuela A, Martínez-Ruiz R, Sánchez-Romero I, Asensio Á, Múñez E, and Plaza A
- Subjects
- Aged, Aged, 80 and over, Antibodies, Bacterial immunology, Antigens, Bacterial immunology, Bacterial Proteins immunology, Comorbidity, Female, Humans, Immunoglobulin A immunology, Immunoglobulin G immunology, Immunoglobulin M immunology, Male, Middle Aged, Recurrence, Spain, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium Infections microbiology, Host-Pathogen Interactions immunology, Immunity, Humoral
- Abstract
Background: Determination of the humoral response to Clostridioides difficile (CD) toxins could be of great value in the management of patients with CD infection (CDI)., Methods: A prospective study was conducted on the clinical characteristics and humoral response in patients with CDI. Determination of ELISA IgG CD anti-toxin B (tgcBiomics, Germany) was performed. The following dilutions were planned for each patient, 1:100, 1: 200, 1: 400, 1: 800: 1: 1600. A significant concentration of antibody was considered to be present in each dilution if an optical density 0.2 units higher than the negative control of the technique was evident., Results: Eighty-five patients were included during the study period, November 2018-February 2020. The median age was 73 years (interquartile range: 62.5-85 years), with female predominance (45 patients, 52.9%). Thirty-nine patients (45.9%) had a severe infection. Seven patients (8.2%) had suffered an episode of CDI in the previous three months. Seventeen patients (20%) had one or more recurrent episodes during the three-month follow-up: No patient died during admission or required surgery for severe-complicated infection. The incidence of recurrence in patients with no antibody detected at 1:400 dilution was 25.4% (16 patients) while it was 4.3% (one patient) in patients with antibody present at that dilution (p = 0.03). Liver cirrhosis was associated with higher humoral response against CD., Conclusions: Antibodies IgG CD anti-toxin B detection at a dilution of 1:400, using a B ELISA technique, effectively identified patients at increased risk of recurrence. This information could help assist in the management of patients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Antonio Ramos-Martínez has received honoraria for lecturing activities and funding for conference attendance from MSD, Astellas, ERN and Angelini., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
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- 2021
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27. Toward the development of defined microbial therapeutics.
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Li Y and Honda K
- Subjects
- Animals, Clostridium Infections immunology, Humans, Phenotype, Clostridium Infections therapy, Fecal Microbiota Transplantation, Gastrointestinal Microbiome immunology
- Abstract
The collection of micro-organisms living in the mammalian gastrointestinal tract, termed the gut microbiota, has been shown to have profound impacts on host health and increasingly is regarded as a viable therapeutic target. Clinical studies of fecal microbiota transplantation have demonstrated potential efficacy of microbiota-based therapies for diseases including Clostridioides difficile infections, inflammatory bowel disease, graft-versus-host disease and cancer. However, the lack of understanding of the active ingredients and potential risks of such therapies pose challenges for clinical application. Meanwhile, efforts are being made to identify effector microbes directly associated with a given phenotype, to establish causality and to devise well-characterized microbial therapeutics for clinical use. Strategies based on defined microbial components will likely enhance the potential of microbiota-targeted therapies., (© The Japanese Society for Immunology. 2021. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2021
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28. A Multi-Factorial Observational Study on Sequential Fecal Microbiota Transplant in Patients with Medically Refractory Clostridioides difficile Infection.
- Author
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Monaghan TM, Duggal NA, Rosati E, Griffin R, Hughes J, Roach B, Yang DY, Wang C, Wong K, Saxinger L, Pučić-Baković M, Vučković F, Klicek F, Lauc G, Tighe P, Mullish BH, Blanco JM, McDonald JAK, Marchesi JR, Xue N, Dottorini T, Acharjee A, Franke A, Li Y, Wong GK, Polytarchou C, Yau TO, Christodoulou N, Hatziapostolou M, Wang M, Russell LA, and Kao DH
- Subjects
- Aged, Aged, 80 and over, Animals, Antibodies, Neutralizing metabolism, Bacterial Toxins immunology, Chlorocebus aethiops, Clostridium Infections immunology, Clostridium Infections microbiology, Cluster Analysis, Feces microbiology, Female, Gastrointestinal Microbiome, Genomics, Humans, Immunosenescence, Male, Middle Aged, Phylogeny, Receptors, Antigen, T-Cell metabolism, Time Factors, Treatment Outcome, Vero Cells, Clostridium Infections therapy, Fecal Microbiota Transplantation
- Abstract
Fecal microbiota transplantation (FMT) is highly effective in recurrent Clostridioides difficile infection (CDI); increasing evidence supports FMT in severe or fulminant Clostridioides difficile infection (SFCDI). However, the multifactorial mechanisms that underpin the efficacy of FMT are not fully understood. Systems biology approaches using high-throughput technologies may help with mechanistic dissection of host-microbial interactions. Here, we have undertaken a deep phenomics study on four adults receiving sequential FMT for SFCDI, in which we performed a longitudinal, integrative analysis of multiple host factors and intestinal microbiome changes. Stool samples were profiled for changes in gut microbiota and metabolites and blood samples for alterations in targeted epigenomic, metabonomic, glycomic, immune proteomic, immunophenotyping, immune functional assays, and T-cell receptor (TCR) repertoires, respectively. We characterised temporal trajectories in gut microbial and host immunometabolic data sets in three responders and one non-responder to sequential FMT. A total of 562 features were used for analysis, of which 78 features were identified, which differed between the responders and the non-responder. The observed dynamic phenotypic changes may potentially suggest immunosenescent signals in the non-responder and may help to underpin the mechanisms accompanying successful FMT, although our study is limited by a small sample size and significant heterogeneity in patient baseline characteristics. Our multi-omics integrative longitudinal analytical approach extends the knowledge regarding mechanisms of efficacy of FMT and highlights preliminary novel signatures, which should be validated in larger studies.
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- 2021
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29. Blockade of T helper 17 cell function ameliorates recurrent Clostridioides difficile infection in mice.
- Author
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Wang S, Deng W, Li F, Chen YE, and Wang PU
- Subjects
- Animals, Apoptosis, Cell Line, Tumor, Clostridium Infections metabolism, Clostridium Infections pathology, Colon drug effects, Colon pathology, Disease Models, Animal, Intestinal Mucosa metabolism, Intestinal Mucosa microbiology, Intestinal Mucosa pathology, Janus Kinase 2 metabolism, Male, Mice, Inbred C57BL, Nuclear Receptor Subfamily 1, Group F, Member 3 antagonists & inhibitors, Recurrence, STAT3 Transcription Factor metabolism, Sulfonamides pharmacology, Sulfonamides therapeutic use, Th17 Cells immunology, Thiazoles pharmacology, Thiazoles therapeutic use, Mice, Clostridium Infections drug therapy, Clostridium Infections immunology, Th17 Cells metabolism
- Abstract
Clostridioides difficile infection (CDI) is a common infection of the gastrointestinal tract. Typically, 20%-30% of CDI patients experience recurrent C.difficile infection (RCDI). Although the role of Th17 in infectious and inflammatory diseases including CDI has gained attention, reports on the correlation between Th17 and RCDI are scarce. In this study, CDI and RCDI mice models were challenged with C. difficile. Serum lactic acid dehydrogenase, inflammatory factor levels, reverse transcriptase-polymerase chain reaction, western blot analysis, hematoxylin and eosin staining, immunohistochemistry, flow cytometry analysis, and enzyme-linked immunosorbent assay were performed on the CDI, RCDI, and control group mice. The results showed more serious clinical manifestations in the RCDI group compared with those in the CDI group. More severe gut barrier disruption and higher degree of microbiota translocation were observed in the RCDI group compared with those in the CDI group. Moreover, extremely severe apoptosis was observed in HCT-116 cells incubated with the serum from RCDI mice model. In addition, higher levels of Th17 and IL-17 were detected in the blood or serum from the RCDI mouse model. Treatment with RORγt small molecule inhibitor SR1001 increased the expression of occludin, decreased the apoptotic rate of HCT-116 cells, and decreased the concentrations of Th17 and IL-17. Concisely, Th17 and IL-17 are potential indicators of RCDI and may serve as therapeutic targets for RCDI treatment. This study lays the foundation for future research on RCDI diagnosis and treatment., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2021
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30. The Murine Neonatal Fc Receptor Is Required for Transport of Immunization-Induced C. difficile-Specific IgG to the Gut and Protection against Disease but Does Not Affect Disease Susceptibility.
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Amadou Amani S, Lang GA, Ballard JD, and Lang ML
- Subjects
- Animals, Antibodies, Monoclonal immunology, Antibodies, Neutralizing immunology, Antitoxins immunology, Bacterial Toxins immunology, Clostridium Infections microbiology, Disease Susceptibility microbiology, Enterotoxins immunology, Female, Immunity immunology, Immunization methods, Male, Mice, Mice, Inbred C57BL, Vaccination methods, Clostridioides difficile immunology, Clostridium Infections immunology, Digestive System immunology, Digestive System microbiology, Disease Susceptibility immunology, Histocompatibility Antigens Class I immunology, Immunoglobulin G immunology, Receptors, Fc immunology
- Abstract
The pathology associated with Clostridioides difficile disease is caused in large part by TcdB, an intracellular bacterial toxin that inactivates small GTPases. Despite C. difficile causing enteric disease, antitoxin IgG is a clear correlate of protection against infection-associated pathology. Immunization with TcdB-based immunogens or passive transfer of monoclonal antibodies specific for the TcdB carboxy-terminal domain (CTD) confers protection following C. difficile infection. Whether the mechanism by which circulating IgG is delivered to the gut depends on specific receptor-mediated transport or is solely reflective of infection-induced damage to the gut remains unclear. Here, we tested the hypothesis that neonatal Fc receptor (FcRn) is required for the delivery of systemic TcdB-specific IgG to the gut and protection against C. difficile-associated pathology. FcRn-expressing mice and FcRn-deficient littermates were immunized subcutaneously with Alhydrogel adjuvant-adsorbed CTD before challenge with live C. difficile spores. FcRn was required for the delivery of systemic TcdB-specific IgG to the gut and for vaccine-induced protection against C. difficile-associated disease. The lack of FcRn expression had minimal effects on the composition of the gut microbiome and did not affect susceptibility to C. difficile infection in nonimmunized mice. In further experiments, intraperitoneal injection of immune sera in FcRn-deficient mice led to the transport of protective IgG to the gut independently of infection, confirming a reported method of bypassing the FcRn. Our results reveal an FcRn-dependent mechanism by which systemic immunization-induced IgG protects the gut during enteric C. difficile infection. These findings may be beneficial for the targeting of C. difficile-specific IgG to the gut.
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- 2021
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31. Measurement over 1 Year of Neutralizing Antibodies in Cattle Immunized with Trivalent Vaccines Recombinant Alpha, Beta and Epsilon of Clostridium perfringens .
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Galvão CC, Barbosa JD, Oliveira CMC, Otaka DY, Silva PRO, Ferreira MRA, Moreira Júnior C, Conceição FR, and Salvarani FM
- Subjects
- Animals, Antibodies, Neutralizing immunology, Bacterial Toxins toxicity, Bacterial Vaccines administration & dosage, Bacterial Vaccines immunology, Brazil, Cattle, Cattle Diseases blood, Cattle Diseases microbiology, Clostridium Infections veterinary, Recombinant Proteins administration & dosage, Antibodies, Neutralizing blood, Bacterial Toxins immunology, Cattle Diseases immunology, Cattle Diseases prevention & control, Clostridium Infections immunology, Clostridium Infections prevention & control, Clostridium perfringens immunology, Recombinant Proteins immunology
- Abstract
The alpha (CPA), beta (CPB) and epsilon (ETX) toxins of Clostridium perfringens are responsible for causing diseases that are difficult to eradicate and have lethal potential in production animals. Vaccination of herds is still the best control strategy. Recombinant clostridial vaccines have shown good success at inducing neutralizing antibody titers and appear to be a viable alternative to the conventional production of commercial clostridial toxoids. Research is still needed on the longevity of the humoral immune response induced by recombinant proteins in immunized animals, preferably in target species. The objective of this study was to measure the humoral immune response of cattle immunized with trivalent vaccines containing the recombinant proteins alpha (rCPA), beta (rCPB) and epsilon (rETX) of C. perfringens produced in Escherichia coli at three different concentrations (100, 200, and 400 µg) of each protein for 12 months. The recombinant vaccines containing 200 (RV2) and 400 µg (RV3) yielded statistically similar results at 56 days. They performed better throughout the study period because they induced higher neutralizing antibody titers and were detectable for up to 150 and 180 days, respectively. Regarding industrial-scale production, RV2 would be the most economical and viable formulation as it achieved results similar to RV3 at half the concentration of recombinant proteins in its formulation. However, none of the vaccines tested induced the production of detectable antibody titers on day 365 of the experiment, the time of revaccination typically recommended in vaccination protocols. Thus, reiterating the need for research in the field of vaccinology to achieve greater longevity of the humoral immune response against these clostridial toxins in animals, in addition to the need to discuss the vaccine schedules and protocols adopted in cattle production.
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- 2021
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32. Clostridioides difficile spores stimulate inflammatory cytokine responses and induce cytotoxicity in macrophages.
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Chiu PJ, Rathod J, Hong YP, Tsai PJ, Hung YP, Ko WC, Chen JW, Paredes-Sabja D, and Huang IH
- Subjects
- Animals, Bacterial Toxins immunology, Clostridioides difficile genetics, Clostridium Infections genetics, Clostridium Infections microbiology, Cytokines genetics, Humans, Macrophages microbiology, Mice, RAW 264.7 Cells, Spores, Bacterial genetics, Clostridioides difficile immunology, Clostridium Infections immunology, Cytokines immunology, Macrophages immunology, Spores, Bacterial immunology
- Abstract
Clostridioides difficile is a gram-positive, spore-forming anaerobic bacterium, and the leading cause of antibiotic-associated diarrhea worldwide. During C. difficile infection, spores germinate in the presence of bile acids into vegetative cells that subsequently colonize the large intestine and produce toxins. In this study, we demonstrated that C. difficile spores can universally adhere to, and be phagocytosed by, murine macrophages. Only spores from toxigenic strains were able to significantly stimulate the production of inflammatory cytokines by macrophages and subsequently induce significant cytotoxicity. Spores from the isogenic TcdA and TcdB double mutant induced significantly lower inflammatory cytokines and cytotoxicity in macrophages, and these activities were restored by pre-exposure of the spores to either toxins. These findings suggest that during sporulation, spores might be coated with C. difficile toxins from the environment, which could affect C. difficile pathogenesis in vivo., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
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- 2021
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33. Immunosuppression and Clostridioides (Clostridium) difficile Infection Risk in Metabolic and Bariatric Surgery Patients.
- Author
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Morales-Marroquin E, Xie L, Uppuluri M, Almandoz JP, Cruz-Muñoz N, and Messiah SE
- Subjects
- Adult, Canada epidemiology, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium Infections microbiology, Female, Gastric Bypass statistics & numerical data, Gastroplasty statistics & numerical data, Humans, Laparoscopy adverse effects, Laparoscopy statistics & numerical data, Male, Middle Aged, Obesity, Morbid surgery, Postoperative Complications immunology, Postoperative Complications microbiology, Retrospective Studies, Risk Assessment statistics & numerical data, Risk Factors, United States epidemiology, Clostridium Infections epidemiology, Gastric Bypass adverse effects, Gastroplasty adverse effects, Immunosuppressive Agents adverse effects, Postoperative Complications epidemiology
- Abstract
Background: Immunosuppressant use increases risk of Clostridioides (Clostridium) difficile infection. To date, no studies have analyzed the relationship between immunosuppressant use and C difficile infections after metabolic and bariatric surgery (MBS)., Methods: A retrospective analysis of the 2015-2018 MBSAQIP data was conducted. The MBSAQIP data include information from 854 affiliated practices in the US and Canada. Initial sample size was 760,076 MBS patients. After excluding participants due to missing variables (n = 188,106) and the use of surgical procedures other than Roux-en-Y gastric bypass and sleeve gastroplasty (n = 129,712), final analyses were performed on 442,258 participants. Logistic regression models generated the odds of C difficile infection developing post MBS, according to immunosuppressant status (positive or negative)., Results: Unadjusted logistic regression analysis showed that patients using immunosuppressants were 95% more likely to have postoperative C difficile infection (odds ratio 1.945; 95% CI, 1.230 to 3.075; p < 0.001) vs MBS patients not taking immunosuppressants. After adjusting for age, sex, ethnicity, preoperative BMI, diabetes status, and surgical procedure type, the association remained unaffected (adjusted odds ratio 1.956; 95% CI, 1.236 to 3.095; p < 0.01). Patients who completed the laparoscopic Roux-en-Y gastric bypass procedure had more than double the odds of C difficile infection developing compared with those who completed the laparoscopic sleeve gastrectomy procedure (odds ratio 2.183; 95% CI, 1.842 to 2.587; p < 0.0001)., Conclusions: Our results using a population-based sample of MBS patients showed that those taking immunosuppressants have a significantly higher risk of developing Clostridioides (Clostridium) difficile infection postoperatively. These findings suggest that patients using immunosuppressants should be closely monitored both pre and post procedure., (Copyright © 2021 American College of Surgeons. Published by Elsevier Inc. All rights reserved.)
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- 2021
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34. Assessment of Immune Response and Efficacy of Essential Oils Application on Controlling Necrotic Enteritis Induced by Clostridium perfringens in Broiler Chickens.
- Author
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Gharaibeh MH, Khalifeh MS, Nawasreh AN, Hananeh WM, and Awawdeh MS
- Subjects
- Animal Feed, Animals, Clostridium Infections microbiology, Clostridium Infections veterinary, Enteritis pathology, Enteritis veterinary, Microbial Sensitivity Tests, Necrosis, Oils, Volatile chemistry, Pilot Projects, Poultry Diseases microbiology, Treatment Outcome, Amoxicillin administration & dosage, Anti-Bacterial Agents administration & dosage, Chickens microbiology, Clostridium Infections drug therapy, Clostridium Infections immunology, Clostridium perfringens drug effects, Enteritis drug therapy, Enteritis immunology, Immunity, Oils, Volatile administration & dosage, Poultry Diseases drug therapy, Poultry Diseases immunology
- Abstract
Necrotic enteritis (NE) caused by Clostridium perfringens is one of the most important enteric diseases in poultry. The antibacterial activity of two different essential oil (EO) blends against C. perfringens was investigated both in vitro and in vivo. Additionally, the immunological response to EO treatment was assessed. In the in vitro study, the antibacterial activity of EO formulas and commonly used antibiotics was evaluated against C. perfringens using disk diffusion assay, minimum inhibitory concentration (MIC) assay, and minimum bactericidal concentration (MBC) assay. In the in vivo study, NE experimental infection was performed on 440 Ross broiler chicks at 19 days of age for 4 continuous days. The chicks were treated with either EOs or amoxicillin at 22 days of age for 5 continuous days. One day after the end of treatment, the birds' performance was evaluated by calculating the feed conversion ratio. Serum samples from 120 birds were collected to measure the levels of IL-1β, IFN-γ, IL-8, IL-10, and IL-17. After that, all birds were slaughtered, and their small intestines were subjected to gross and histopathological evaluation. In addition, bacterial counts in the small intestines were evaluated. In the in vitro study, EOs showed higher antimicrobial activities in comparison with antibiotics against C. perfringens . In the in vivo study, birds treated with EOs showed a significant decrease in bacterial counts, a significant decrease in intestinal lesions, and a significant improvement in performance compared with untreated birds ( p < 0.05). Moreover, treating birds with EOs directed the immune system toward an anti-inflammatory pathway. None of the treated birds died due to NE compared with the 10% mortality rate in untreated birds. In conclusion, EOs might be an effective and safe alternative to antibiotics in the treatment of chicken NE.
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- 2021
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35. The Role of Toll-Like Receptor-2 in Clostridioides difficile Infection: Evidence From a Mouse Model and Clinical Patients.
- Author
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Lai YH, Tsai BY, Hsu CY, Chen YH, Chou PH, Chen YL, Liu HC, Ko WC, Tsai PJ, and Hung YP
- Subjects
- Animals, Asian People genetics, Clostridioides difficile, Clostridium Infections genetics, Clostridium Infections pathology, Colon immunology, Colon pathology, Disease Models, Animal, Female, Genotype, Humans, Male, Mice, Knockout, Polymorphism, Single Nucleotide, Toll-Like Receptor 2 genetics, Toll-Like Receptor 4 genetics, Toll-Like Receptor 4 immunology, Mice, Clostridium Infections immunology, Toll-Like Receptor 2 immunology
- Abstract
Background: Clostridioides difficile is the leading cause of nosocomial infectious diarrhea. Toll-like receptors (TLRs) are the major components of innate immunity that sense pathogens. The relationship between TLRs and C. difficile infection (CDI) was analyzed in clinical patients and a mouse model., Materials and Methods: A prospective investigation was conducted in medical wards of Tainan Hospital, Ministry of Health and Welfare, Tainan, Taiwan, from January 2011 to January 2013. Adult patients were followed up for the development of CDI. Single nucleotide polymorphisms (SNPs) of TLR2 and TLR4 were analyzed to assess the relationship between genetic polymorphisms and the development of CDI. A mouse model of CDI was used to investigate the pathogenic role of TLRs in CDI, TLR2 and TLR4 knockout (Tlr2-/- and Tlr4-/-) mice., Results: In the prospective study, 556 patients were enrolled, and 6.5% (36) of patients, accounting for 3.59 episodes per 1000 patient-days, developed CDI. Of 539 patients with available blood samples, the TLR2 rs3804099 polymorphism was more often noted in those with CDI than in those without CDI (64.5% vs. 46.1%; P = 0.046) but was not significant in multivariate analysis. Because the TLR2 rs3804099 polymorphism was moderately associated with CDI, the role of TLR2 and TLR4 was further evaluated in a mouse model. Both Tlr2-/- and Tlr4-/- mice showed more severe CDI disease than wild-type mice in terms of body weight change and fecal content five days after oral challenge with C. difficile . Furthermore, Tlr2-/- mice suffered from more severe disease than Tlr4-/- mice, as evidenced by stool consistency, cecum weight, and survival rate., Conclusion: The TLR2 rs3804099 polymorphism is marginally associated with the development of CDI, and the pathogenic role of TLR2 is further supported by a mouse model., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Lai, Tsai, Hsu, Chen, Chou, Chen, Liu, Ko, Tsai and Hung.)
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- 2021
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36. In silico designing of vaccine candidate against Clostridium difficile.
- Author
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Basak S, Deb D, Narsaria U, Kar T, Castiglione F, Sanyal I, Bade PD, and Srivastava AP
- Subjects
- Computational Biology methods, Escherichia coli metabolism, Humans, Bacterial Vaccines immunology, Bacterial Vaccines therapeutic use, Clostridioides difficile immunology, Clostridioides difficile pathogenicity, Clostridium Infections drug therapy, Clostridium Infections immunology
- Abstract
Clostridium difficile is a spore-forming gram-positive bacterium, recognized as the primary cause of antibiotic-associated nosocomial diarrhoea. Clostridium difficile infection (CDI) has emerged as a major health-associated infection with increased incidence and hospitalization over the years with high mortality rates. Contamination and infection occur after ingestion of vegetative spores, which germinate in the gastro-intestinal tract. The surface layer protein and flagellar proteins are responsible for the bacterial colonization while the spore coat protein, is associated with spore colonization. Both these factors are the main concern of the recurrence of CDI in hospitalized patients. In this study, the CotE, SlpA and FliC proteins are chosen to form a multivalent, multi-epitopic, chimeric vaccine candidate using the immunoinformatics approach. The overall reliability of the candidate vaccine was validated in silico and the molecular dynamics simulation verified the stability of the vaccine designed. Docking studies showed stable vaccine interactions with Toll-Like Receptors of innate immune cells and MHC receptors. In silico codon optimization of the vaccine and its insertion in the cloning vector indicates a competent expression of the modelled vaccine in E. coli expression system. An in silico immune simulation system evaluated the effectiveness of the candidate vaccine to trigger a protective immune response., (© 2021. The Author(s).)
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- 2021
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37. Immunomodulatory effect of short-term supplementation with Bacillus toyonensis BCT-7112 T and Saccharomyces boulardii CNCM I-745 in sheep vaccinated with Clostridium chauvoei.
- Author
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Santos FDS, Maubrigades LR, Gonçalves VS, Alves Ferreira MR, Brasil CL, Cunha RC, Conceição FR, and Leite FPL
- Subjects
- Animals, Antibodies, Bacterial immunology, Clostridium Infections immunology, Clostridium Infections prevention & control, Female, Immunoglobulin G immunology, Immunomodulation, Interferon-gamma genetics, Interleukin-2 genetics, Probiotics administration & dosage, Proto-Oncogene Proteins c-bcl-6 genetics, Sheep, Sheep Diseases immunology, Transcription, Genetic, Bacillus immunology, Bacterial Vaccines immunology, Clostridium Infections veterinary, Clostridium chauvoei immunology, Saccharomyces boulardii immunology, Sheep Diseases prevention & control
- Abstract
The bacterium Clostridium chauvoei is the causative agent of blackleg in livestock, and vaccination is the most effective means of prevention. The aim of this study was to assess the effect of short-term supplementation with Bacillus toyonensis and Saccharomyces boulardii on the immune response to a C. chauvoei vaccine in sheep. Sheep were vaccinated subcutaneously on day 0 and received a booster dose on day 21, with 2 mL of a commercial vaccine formulated with inactivated C. chauvoei bacterin adsorbed on aluminum hydroxide. Probiotics were orally administered B. toyonensis (3 × 10
8 cfu) and S. boulardii (3 × 108 cfu) over five days prior to the first and second doses of the vaccine. Sheep supplemented with B. toyonensis and S. boulardii showed significantly higher specific IgG, IgG1, and IgG2 titers (P<0.05), with approximately 24- and 14-fold increases in total IgG levels, respectively, than the nonsupplemented group. Peripheral blood mononuclear cells from the supplemented group had increased mRNA transcription levels of the IFN-γ, IL2, and Bcl6 genes. These results demonstrate an adjuvant effect of short-term supplementation with B. toyonensis and S. boulardii on the immune response against the C. chauvoei vaccine in sheep., (Copyright © 2021 Elsevier B.V. All rights reserved.)- Published
- 2021
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38. Intestinal Inflammation and Altered Gut Microbiota Associated with Inflammatory Bowel Disease Render Mice Susceptible to Clostridioides difficile Colonization and Infection.
- Author
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Abernathy-Close L, Barron MR, George JM, Dieterle MG, Vendrov KC, Bergin IL, and Young VB
- Subjects
- Animals, Clostridium Infections immunology, Disease Models, Animal, Disease Susceptibility, Female, Intestines microbiology, Intestines pathology, Male, Mice, Mice, Inbred C57BL, Clostridioides difficile pathogenicity, Clostridium Infections etiology, Gastrointestinal Microbiome, Inflammation complications, Inflammatory Bowel Diseases complications, Inflammatory Bowel Diseases microbiology, Intestines immunology
- Abstract
Clostridioides difficile is a noteworthy pathogen in patients with inflammatory bowel disease (IBD). Patients with IBD who develop concurrent C. difficile infection (CDI) experience increased morbidity and mortality. IBD is associated with intestinal inflammation and alterations of the gut microbiota, both of which can diminish colonization resistance to C. difficile. Here, we describe the development of a mouse model to explore the role that IBD-induced changes of the gut microbiome play in susceptibility to C. difficile. Helicobacter hepaticus, a normal member of the mouse gut microbiota, triggers pathological inflammation in the distal intestine akin to human IBD in mice that lack intact interleukin 10 (IL-10) signaling. We demonstrate that mice with H. hepaticus-induced IBD were susceptible to C. difficile colonization in the absence of other perturbations, such as antibiotic treatment. Concomitant IBD and CDI were associated with significantly worse disease than observed in animals with colitis alone. Development of IBD resulted in a distinct intestinal microbiota community compared to that of non-IBD controls. Inflammation played a critical role in the susceptibility of animals with IBD to C. difficile colonization, as mice colonized with an isogenic mutant of H. hepaticus that triggers an attenuated intestinal inflammation maintained full colonization resistance. These studies with a novel mouse model of IBD and CDI emphasize the importance of host responses and alterations of the gut microbiota in susceptibility to C. difficile colonization and infection in the setting of IBD. IMPORTANCE The incidence of C. difficile infection (CDI) has increased significantly among patients with IBD, independently of antibiotic use, yet the relationship between IBD and increased risk for CDI remains to be understood. Our study sought to describe and utilize an antibiotic-independent mouse model to specifically explore the relationship between the IBD-associated gut and susceptibility to C. difficile colonization and CDI development. We demonstrate that the development of IBD is sufficient to render mice susceptible to C. difficile colonization and results in significantly worse disease than IBD alone. Furthermore, this model requires IBD-induced inflammation to overcome colonization resistance to C. difficile. This model recapitulates human IBD and CDI comorbidity and will aid in developing new clinical approaches to predict, diagnose, and treat C. difficile infection in the IBD population.
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- 2021
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39. Mechanisms of Colonization Resistance Against Clostridioides difficile.
- Author
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Pike CM and Theriot CM
- Subjects
- Animals, Anti-Bacterial Agents immunology, Anti-Bacterial Agents metabolism, Anti-Bacterial Agents therapeutic use, Antibiosis, Bile Acids and Salts immunology, Bile Acids and Salts metabolism, Clostridioides difficile drug effects, Clostridioides difficile metabolism, Clostridium Infections drug therapy, Clostridium Infections microbiology, Fatty Acids, Volatile immunology, Fatty Acids, Volatile metabolism, Gastrointestinal Microbiome drug effects, Gastrointestinal Microbiome physiology, Humans, Nutrients immunology, Nutrients metabolism, Clostridioides difficile growth & development, Clostridium Infections immunology
- Abstract
Clostridioides difficile is an urgent antimicrobial-resistant bacterium, causing mild to moderate and sometimes life-threatening disease. Commensal gut microbes are critical for providing colonization resistance against C difficile and can be leveraged as non-antibiotic alternative therapeutics for the prevention and treatment of C difficile infection., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)
- Published
- 2021
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40. Identification of linear epitopes on the flagellar proteins of Clostridioides difficile.
- Author
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Razim A, Pacyga K, Naporowski P, Martynowski D, Szuba A, Gamian A, and Górska S
- Subjects
- Amino Acid Sequence genetics, Animals, Antibodies, Bacterial immunology, Bacterial Proteins immunology, Bacterial Vaccines genetics, Bacterial Vaccines immunology, Clostridioides genetics, Clostridioides immunology, Clostridioides difficile pathogenicity, Clostridium Infections immunology, Epitopes immunology, Flagellin immunology, Humans, Rabbits, Sequence Alignment methods, Clostridioides difficile immunology, Clostridium Infections diagnosis, Flagella immunology
- Abstract
Clostridioides difficile (C. difficile) is an opportunistic anaerobic bacterium that causes severe diseases of the digestive tract of humans and animals. One of the possible methods of preventing C. difficile infection is to develop a vaccine. The most promising candidates for vaccine antigens are the proteins involved in the adhesion phenomena. Among them, the FliC and FliD are considered to be suitable candidates. In this paper, the FliC and FliD protein polypeptide epitopes were mapped in silico and by using PEPSCAN procedure. We identified four promising epitopes:
117 QRMRTLS123 ,205 MSKAG209 of FliC and226 NKVAS230 ,306 TTKKPKD312 of FliD protein. We showed that117 QRMRTLS123 sequence is not only located in TLR5-binding and activating region, as previously shown, but forms an epitope recognized by C. difficile-infected patients' antibodies.205 MSKAG209 is a C. difficile-unique, immunogenic sequence that forms an exposed epitope on the polymerized flagella structure which makes it a suitable vaccine antigen.226 NKVAS230 and306 TTKKPKD312 are well exposed and possess potential protective properties according to VaxiJen analysis. Our results open the possibility to use these epitopes as suitable anti-C. difficile vaccine antigens.- Published
- 2021
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41. Loss of Interleukin-10 (IL-10) Signaling Promotes IL-22-Dependent Host Defenses against Acute Clostridioides difficile Infection.
- Author
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Cribas ES, Denny JE, Maslanka JR, and Abt MC
- Subjects
- Animals, Clostridium Infections immunology, Cytokines metabolism, Disease Models, Animal, Disease Susceptibility, Immunity, Innate, Inflammation Mediators metabolism, Mice, Mice, Knockout, Interleukin-22, Clostridioides difficile physiology, Clostridium Infections metabolism, Clostridium Infections microbiology, Host-Pathogen Interactions immunology, Interleukin-10 metabolism, Interleukins metabolism, Signal Transduction
- Abstract
Infection with the bacterial pathogen Clostridioides difficile causes severe damage to the intestinal epithelium that elicits a robust inflammatory response. Markers of intestinal inflammation accurately predict clinical disease, however, the extent to which host-derived proinflammatory mediators drive pathogenesis versus promote host protective mechanisms remains elusive. In this report, we employed Il10
-/- mice as a model of spontaneous colitis to examine the impact of constitutive intestinal immune activation, independent of infection, on C. difficile disease pathogenesis. Upon C. difficile challenge, Il10-/- mice exhibited significantly decreased morbidity and mortality compared to littermate Il10 heterozygote ( Il10HET ) control mice, despite a comparable C. difficile burden, innate immune response, and microbiota composition following infection. Similarly, antibody-mediated blockade of interleukin-10 (IL-10) signaling in wild-type C57BL/6 mice conveyed a survival advantage if initiated 3 weeks prior to infection. In contrast, no advantage was observed if blockade was initiated on the day of infection, suggesting that the constitutive activation of inflammatory defense pathways prior to infection mediated host protection. IL-22, a cytokine critical in mounting a protective response against C. difficile infection, was elevated in the intestine of uninfected, antibiotic-treated Il10-/- mice, and genetic ablation of the IL-22 signaling pathway in Il10-/- mice negated the survival advantage following C. difficile challenge. Collectively, these data demonstrate that constitutive loss of IL-10 signaling, via genetic ablation or antibody blockade, enhances IL-22-dependent host defense mechanisms to limit C. difficile pathogenesis., (Copyright © 2021 American Society for Microbiology.)- Published
- 2021
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42. The Butyrate-Producing Bacterium Clostridium butyricum Suppresses Clostridioides difficile Infection via Neutrophil- and Antimicrobial Cytokine-Dependent but GPR43/109a-Independent Mechanisms.
- Author
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Hayashi A, Nagao-Kitamoto H, Kitamoto S, Kim CH, and Kamada N
- Subjects
- Animals, Butyrates metabolism, Interferon-gamma metabolism, Interleukin-17 metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Receptors, G-Protein-Coupled genetics, alpha-Defensins metabolism, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium butyricum physiology, Colon immunology, Neutrophils immunology, Receptors, G-Protein-Coupled metabolism
- Abstract
Short-chain fatty acids, such as butyrate, are major gut microbial metabolites that are beneficial for gastrointestinal health. Clostridium butyricum MIYAIRI588 (CBM588) is a bacterium that produces a robust amount of butyrate and therefore has been used as a live biotherapeutic probiotic in clinical settings. Clostridioides difficile causes life-threatening diarrhea and colitis. The gut resident microbiota plays a critical role in the prevention of C. difficile infection (CDI), as the disruption of the healthy microbiota by antibiotics greatly increases the risk for CDI. We report that CBM588 treatment in mice significantly improved clinical symptoms associated with CDI and increased the number of neutrophils and Th1 and Th17 cells in the colonic lamina propria in the early phase of CDI. The protective effect of CBM588 was abolished when neutrophils, IFN-γ, or IL-17A were depleted, suggesting that induction of the immune reactants is required to elicit the protective effect of the probiotic. The administration of tributyrin, which elevates the concentration of butyrate in the colon, also increased the number of neutrophils in the colonic lamina propria, indicating that butyrate is a potent booster of neutrophil activity during infection. However, GPR43 and GPR109a, two G protein-coupled receptors activated by butyrate, were dispensable for the protective effect of CBM588. These results indicate that CBM588 and butyrate suppress CDI, in part by boosting antimicrobial innate and cytokine-mediated immunity., (Copyright © 2021 by The American Association of Immunologists, Inc.)
- Published
- 2021
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43. Fusobacteriumnucleatum Adheres to Clostridioides difficile via the RadD Adhesin to Enhance Biofilm Formation in Intestinal Mucus.
- Author
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Engevik MA, Danhof HA, Auchtung J, Endres BT, Ruan W, Bassères E, Engevik AC, Wu Q, Nicholson M, Luna RA, Garey KW, Crawford SE, Estes MK, Lux R, Yacyshyn MB, Yacyshyn B, Savidge T, Britton RA, and Versalovic J
- Subjects
- Adhesins, Bacterial genetics, Bacterial Adhesion immunology, Biofilms, Bioreactors microbiology, Clostridioides difficile genetics, Clostridioides difficile immunology, Clostridioides difficile metabolism, Clostridium Infections microbiology, Feces microbiology, Flagella genetics, Flagella metabolism, Fusobacterium nucleatum metabolism, HT29 Cells, Humans, Intestinal Mucosa immunology, Intestinal Mucosa metabolism, Intestinal Mucosa microbiology, Intestinal Mucosa pathology, Mucin-2 metabolism, Adhesins, Bacterial metabolism, Clostridioides difficile pathogenicity, Clostridium Infections immunology, Fusobacterium nucleatum immunology, Gastrointestinal Microbiome immunology
- Abstract
Background & Aims: Although Clostridioides difficile infection (CDI) is known to involve the disruption of the gut microbiota, little is understood regarding how mucus-associated microbes interact with C difficile. We hypothesized that select mucus-associated bacteria would promote C difficile colonization and biofilm formation., Methods: To create a model of the human intestinal mucus layer and gut microbiota, we used bioreactors inoculated with healthy human feces, treated with clindamycin and infected with C difficile with the addition of human MUC2-coated coverslips., Results: C difficile was found to colonize and form biofilms on MUC2-coated coverslips, and 16S rRNA sequencing showed a unique biofilm profile with substantial cocolonization with Fusobacterium species. Consistent with our bioreactor data, publicly available data sets and patient stool samples showed that a subset of patients with C difficile infection harbored high levels of Fusobacterium species. We observed colocalization of C difficile and F nucleatum in an aggregation assay using adult patients and stool of pediatric patients with inflammatory bowel disease and in tissue sections of patients with CDI. C difficile strains were found to coaggregate with F nucleatum subspecies in vitro; an effect that was inhibited by blocking or mutating the adhesin RadD on Fusobacterium and removal of flagella on C difficile. Aggregation was shown to be unique between F nucleatum and C difficile, because other gut commensals did not aggregate with C difficile. Addition of F nucleatum also enhanced C difficile biofilm formation and extracellular polysaccharide production., Conclusions: Collectively, these data show a unique interaction of between pathogenic C difficile and F nucleatum in the intestinal mucus layer., (Copyright © 2021 AGA Institute. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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44. Recurrent Clostridioides difficile Infection Is Associated With Impaired T Helper Type 17 Immunity to C difficile Toxin B.
- Author
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Cook L, Rees WD, Wong MQ, Kwok WW, Levings MK, and Steiner TS
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Anti-Bacterial Agents therapeutic use, Case-Control Studies, Clostridioides difficile isolation & purification, Clostridium Infections microbiology, Clostridium Infections therapy, Fecal Microbiota Transplantation, Female, Healthy Volunteers, Humans, Male, Middle Aged, Recurrence, Young Adult, Bacterial Proteins immunology, Bacterial Toxins immunology, Clostridioides difficile immunology, Clostridium Infections immunology, Th17 Cells immunology
- Published
- 2021
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45. A standardised model for stool banking for faecal microbiota transplantation: a consensus report from a multidisciplinary UEG working group.
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Keller JJ, Ooijevaar RE, Hvas CL, Terveer EM, Lieberknecht SC, Högenauer C, Arkkila P, Sokol H, Gridnyev O, Mégraud F, Kump PK, Nakov R, Goldenberg SD, Satokari R, Tkatch S, Sanguinetti M, Cammarota G, Dorofeev A, Gubska O, Ianiro G, Mattila E, Arasaradnam RP, Sarin SK, Sood A, Putignani L, Alric L, Baunwall SMD, Kupcinskas J, Link A, Goorhuis AG, Verspaget HW, Ponsioen C, Hold GL, Tilg H, Kassam Z, Kuijper EJ, Gasbarrini A, Mulder CJJ, Williams HRT, and Vehreschild MJGT
- Subjects
- Age Factors, Biological Specimen Banks standards, Clostridioides difficile, Clostridium Infections immunology, Clostridium Infections therapy, Contraindications, Procedure, Donor Selection, Humans, Immunocompromised Host, Informed Consent, Quality Assurance, Health Care, Recurrence, Specimen Handling, Biological Specimen Banks organization & administration, Fecal Microbiota Transplantation adverse effects, Fecal Microbiota Transplantation methods, Feces
- Abstract
Background: Faecal microbiota transplantation is an emerging therapeutic option, particularly for the treatment of recurrent Clostridioides difficile infection. Stool banks that organise recruitment and screening of faeces donors are being embedded within the regulatory frameworks described in the European Union Tissue and Cells Directive and the technical guide to the quality and safety of tissue and cells for human application, published by the European Council., Objective: Several European and international consensus statements concerning faecal microbiota transplantation have been issued. While these documents provide overall guidance, we aim to provide a detailed description of all processes that relate to the collection, handling and clinical application of human donor stool in this document., Methods: Collaborative subgroups of experts on stool banking drafted concepts for all domains pertaining to stool banking. During a working group meeting in the United European Gastroenterology Week 2019 in Barcelona, these concepts were discussed and finalised to be included in our overall guidance document about faecal microbiota transplantation., Results: A guidance document for all domains pertaining to stool banking was created. This document includes standard operating manuals for several processes involved with stool banking, such as handling of donor material, storage and donor screening., Conclusion: The implementation of faecal microbiota transplantation by stool banks in concordance with our guidance document will enable quality assurance and guarantee the availability of donor faeces preparations for patients., (© 2020 The Authors. United European Gastroenterology Journal published by Wiley Periodicals LLC. on behalf of United European Gastroenterology.)
- Published
- 2021
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46. Protective Effects of Lactobacillus plantarum 16 and Paenibacillus polymyxa 10 Against Clostridium perfringens Infection in Broilers.
- Author
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Gong L, Wang B, Zhou Y, Tang L, Zeng Z, Zhang H, and Li W
- Subjects
- Animals, Chickens immunology, Chickens microbiology, Clostridium Infections immunology, Clostridium Infections microbiology, Clostridium Infections prevention & control, Clostridium perfringens immunology, Lactobacillus plantarum immunology, Paenibacillus polymyxa immunology, Poultry Diseases immunology, Poultry Diseases microbiology, Poultry Diseases prevention & control
- Abstract
This study aimed to investigate the protective effects of Lactobacillus plantarum 16 (Lac16) and Paenibacillus polymyx a 10 (BSC10) against Clostridium perfringens (Cp) infection in broilers. A total of 720 one-day-old chicks were randomly divided into four groups. The control and Cp group were only fed a basal diet, while the two treatment groups received basal diets supplemented with Lac16 (1 × 10
8 cfu·kg-1 ) and BSC10 (1 × 108 cfu·kg-1 ) for 21 days, respectively. On day 1 and days 14 to 20, birds except those in the control group were challenged with 1 × 108 cfu C. perfringens type A strain once a day. The results showed that both Lac16 and BSC10 could ameliorate intestinal structure damage caused by C. perfringens infection. C. perfringens infection induced apoptosis by increasing the expression of Bax and p53 and decreasing Bcl-2 expression and inflammation evidence by higher levels of IFN-γ, IL-6, IL-1β , iNOS , and IL-10 in the ileum mucosa, and NO production in jejunal mucosa, which was reversed by Lac16 and BSC10 treatment except for IL-1β ( P < 0.05). Besides, the two probiotics restored the intestinal microbiota imbalance induced by C. perfringens infection, characterized by the reduced Firmicutes and Proteobacteria and the increased Bacteroidetes at the phyla level and decreased Bacteroides fragilis and Gallibacterium anatis at the genus level. The two probiotics also reversed metabolic pathways of the microbiota in C. perfringens -infected broilers, including B-vitamin biosynthesis, peptidoglycan biosynthesis, and pyruvate fermentation to acetate and lactate II pathway. In conclusion, Lac16 and BSC10 can effectively protect broilers against C. perfringens infection through improved composition and metabolic pathways of the intestinal microbiota, intestinal structure, inflammation, and anti-apoptosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Gong, Wang, Zhou, Tang, Zeng, Zhang and Li.)- Published
- 2021
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47. Host immunity modulates the efficacy of microbiota transplantation for treatment of Clostridioides difficile infection.
- Author
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Littmann ER, Lee JJ, Denny JE, Alam Z, Maslanka JR, Zarin I, Matsuda R, Carter RA, Susac B, Saffern MS, Fett B, Mattei LM, Bittinger K, and Abt MC
- Subjects
- Animals, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes metabolism, Clostridium Infections immunology, Clostridium Infections metabolism, Feces microbiology, Forkhead Transcription Factors metabolism, Homeodomain Proteins metabolism, Inflammation immunology, Inflammation metabolism, Mice, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism, Clostridioides difficile pathogenicity
- Abstract
Fecal microbiota transplantation (FMT) is a successful therapeutic strategy for treating recurrent Clostridioides difficile infection. Despite remarkable efficacy, implementation of FMT therapy is limited and the mechanism of action remains poorly understood. Here, we demonstrate a critical role for the immune system in supporting FMT using a murine C. difficile infection system. Following FMT, Rag1 heterozygote mice resolve C. difficile while littermate Rag1
-/- mice fail to clear the infection. Targeted ablation of adaptive immune cell subsets reveal a necessary role for CD4+ Foxp3+ T-regulatory cells, but not B cells or CD8+ T cells, in FMT-mediated resolution of C. difficile infection. FMT non-responsive mice exhibit exacerbated inflammation, impaired engraftment of the FMT bacterial community and failed restoration of commensal bacteria-derived secondary bile acid metabolites in the large intestine. These data demonstrate that the host's inflammatory immune status can limit the efficacy of microbiota-based therapeutics to treat C. difficile infection.- Published
- 2021
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48. Anti-toxin antibody is not associated with recurrent Clostridium difficile infection.
- Author
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Gilbert J, Leslie J, Putler R, Weiner S, Standke A, Penkevich A, Keidan M, Young VB, and Rao K
- Subjects
- Adult, Aged, Antibodies, Bacterial blood, Antibodies, Neutralizing immunology, Bacterial Proteins immunology, Clostridioides difficile, Clostridium Infections diagnosis, Enzyme-Linked Immunosorbent Assay, Female, Humans, Immunoglobulin G blood, Immunoglobulin G immunology, Logistic Models, Male, Middle Aged, Polymerase Chain Reaction, Prognosis, Prospective Studies, Recurrence, Retrospective Studies, Ribotyping, Risk Factors, Antibodies, Bacterial immunology, Bacterial Toxins immunology, Clostridium Infections immunology
- Abstract
Clostridium difficile infection (CDI) recurs in ∼20% of patients. Prior studies indicated that antibody responses directed against the C. difficile toxins A and B were potentially associated with lower risk of recurrent CDI. Here we tested the hypothesis that circulating anti-toxin IgG antibody levels associate with reduced risk of recurrent CDI. A cohort study with prospective enrollment and retrospective data abstraction examined antibody levels in 275 adult patients at the University of Michigan with CDI. We developed an enzyme linked immunosorbent assay to detect IgG antibodies against toxin A and toxin B in sera obtained at the time of diagnosis. Logistic regression examined the relationship between antibody levels and recurrence, and sensitivity tests evaluated for follow-up and survivor biases, history of CDI, and PCR ribotype. Follow-up data were available for 174 subjects, of whom 36 (20.7%) had recurrence. Comparing antibody levels vs. recurrence and CDI history, anti-toxin A levels were similar, while anti-toxin B levels had a greater range of values. In unadjusted analysis, detection of anti-toxin A antibodies, but not anti-toxin B antibodies, associated with an increased risk of recurrence (OR 2.71 [1.06, 8.37], P = .053). Adjusting for confounders weakened this association. The results were the same in sensitivity analyses. We observed a borderline increased risk of recurrence in patients positive for anti-toxin A antibodies, and sensitivity analyses showed this was not simply a reflection of prior exposure status. Future studies are needed to assess how neutralizing antibody or levels after treatment associate with recurrence., Competing Interests: Declaration of competing interest Krishna Rao has consulted for Bio-K + International, Inc. and Roche Molecular Systems, Inc. Dr. Rao holds a grant for an investigator initiated study supported by Merck and Co., Inc. Vincent Young has consulted for Bio-K + International, Inc., Pantheryx, Exarca Pharmaceuticals, and Vedanta., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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49. Clostridioides difficile exploits toxin-mediated inflammation to alter the host nutritional landscape and exclude competitors from the gut microbiota.
- Author
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Fletcher JR, Pike CM, Parsons RJ, Rivera AJ, Foley MH, McLaren MR, Montgomery SA, and Theriot CM
- Subjects
- Animals, Anti-Bacterial Agents adverse effects, Bacterial Proteins genetics, Bacterial Toxins genetics, Bacterial Toxins immunology, Bacteroides drug effects, Bacteroides metabolism, Clostridioides difficile genetics, Clostridioides difficile immunology, Clostridium Infections microbiology, Clostridium Infections pathology, Disease Models, Animal, Extracellular Matrix metabolism, Female, Gastrointestinal Microbiome drug effects, Gene Expression Regulation, Bacterial immunology, Host-Pathogen Interactions genetics, Host-Pathogen Interactions immunology, Humans, Intestinal Mucosa immunology, Intestinal Mucosa microbiology, Intestinal Mucosa pathology, Male, Matrix Metalloproteinases metabolism, Mice, Nutrients metabolism, Proteolysis, RNA, Bacterial genetics, RNA, Bacterial isolation & purification, RNA-Seq, Sigma Factor genetics, Sigma Factor immunology, Transcriptome immunology, Bacterial Proteins metabolism, Bacterial Toxins metabolism, Clostridioides difficile metabolism, Clostridium Infections immunology, Gastrointestinal Microbiome immunology, Sigma Factor metabolism
- Abstract
Clostridioides difficile is a bacterial pathogen that causes a range of clinical disease from mild to moderate diarrhea, pseudomembranous colitis, and toxic megacolon. Typically, C. difficile infections (CDIs) occur after antibiotic treatment, which alters the gut microbiota, decreasing colonization resistance against C. difficile. Disease is mediated by two large toxins and the expression of their genes is induced upon nutrient depletion via the alternative sigma factor TcdR. Here, we use tcdR mutants in two strains of C. difficile and omics to investigate how toxin-induced inflammation alters C. difficile metabolism, tissue gene expression and the gut microbiota, and to determine how inflammation by the host may be beneficial to C. difficile. We show that C. difficile metabolism is significantly different in the face of inflammation, with changes in many carbohydrate and amino acid uptake and utilization pathways. Host gene expression signatures suggest that degradation of collagen and other components of the extracellular matrix by matrix metalloproteinases is a major source of peptides and amino acids that supports C. difficile growth in vivo. Lastly, the inflammation induced by C. difficile toxin activity alters the gut microbiota, excluding members from the genus Bacteroides that are able to utilize the same essential nutrients released from collagen degradation.
- Published
- 2021
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50. MicroRNA-21-5p targets PDCD4 to modulate apoptosis and inflammatory response to Clostridium perfringens beta2 toxin infection in IPEC-J2 cells.
- Author
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Gao X, Huang X, Yang Q, Zhang S, Yan Z, Luo R, Wang P, Wang W, Xie K, and Gun S
- Subjects
- Animals, Apoptosis, Apoptosis Regulatory Proteins genetics, Bacterial Toxins genetics, Cell Line, Cytokines metabolism, HEK293 Cells, Humans, Intestinal Mucosa pathology, NF-kappa B metabolism, Proto-Oncogene Proteins c-bcl-2 metabolism, Signal Transduction, Apoptosis Regulatory Proteins metabolism, Bacterial Toxins metabolism, Clostridium Infections immunology, Clostridium perfringens physiology, Inflammation immunology, Intestinal Mucosa metabolism, MicroRNAs genetics, Swine immunology
- Abstract
Clostridium perfringens (C. perfringens), a toxin-producing enteric pathogen, causes a variety of intestinal infections in humans and animals. C. perfringens beta2 (CPB2) toxin has been considered to be a strong virulence factor for C. perfringens infectious enteric diseases (CPED). Altered levels and functions of microRNA-21-5p (miR-21-5p) have been associated with apoptosis and inflammation response in pathological processes. However, little is known about its functional mechanism in CPED. Here, we found that miR-21-5p expressed in multiple tissues of pig, had a highest level in jejunum, and significantly upregulated in intestinal porcine epithelial cells (IPEC-J2) exposed to CPB2 toxin. Noteworthily, transfection of CPB2-treated IPEC-J2 cells with miR-21-5p mimic increased cell viability and Bcl2 expression, as well as reduced cytotoxicity, apoptosis rates and Bax level. Moreover, overexpression of miR-21-5p significantly suppressed the levels of interleukin (IL)-6, IL-8, TNF-α, IL-1β and nuclear factor-kappa B (NF-κB p65) activity induced by CPB2 toxin, whereas that of the IL-10 was increased in IPEC-J2 cells. On the contrary, transfection of miR-21-5p inhibitor promoted CPB2-induced cell apoptosis and inflammation. Furthermore, we validated that programmed cell death 4 (PDCD4) was strikingly downregulated in CPB2-treated IPEC-J2 cells. PDCD4 exhibited opposing effects to those of miR-21-5p mimic on IPEC-J2 cells, and restoration of PDCD4 expression counteracted the suppressive effect of miR-21-5p on CPB2-induced apoptosis and inflammatory response. Collectively, our findings demonstrated that miR-21-5p was involved in regulating the immune response triggered by CPB2 toxin and contributed to protective effects in CPB2-induced CPED cell model by targeting PDCD4., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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