28 results on '"Péchiné S"'
Search Results
2. Structure and assembly of a Clostridioides difficile spore polar appendage
- Author
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Pereira Fc, Melo Lv, Vultos Td, Péchiné S, Serrano M, Henriques Ao, Antunes W, Isabelle Martin-Verstraete, Brito P, Feliciano C, Janoir C, Couture-Tosi E, Laure Saujet, Jean-François Bruxelle, and Bruno Dupuy
- Subjects
Appendage ,Crystallography ,fungi ,Polar ,Biology - Abstract
Clostridioides difficile, a strict anaerobic spore-former, is the main cause of nosocomial disease associated to antibiotic therapy in adults and a growing concern in the community. Spores are the main infectious, persistence and transmission vehicle. Spore germination occurs in the intestine and the resulting vegetative cells will produce the toxins responsible for the disease symptoms, and spores. During sporulation, a wild type population bifurcates into two main spore morphotypes, with or without a thick exosporium. We show that this bifurcation extends to the formation of spores with a robust polar appendage or spores with a short appendage or that lack this structure. The cysteine-rich CdeM protein localizes to the appendage and around the entire surface of the spore, and is a major structural component of the exosporium, which we show is continuous with the appendage. In aCdeMmutant, when present, the polar appendage is short and disorganized. We show that wild type andcdeMspores with a short or no appendage germinate poorly in response to taurocholate, compared to those with an appendage.cdeMspores of the two types, however, germinate faster than their wild type counterparts. Thus, while the absence of CdeM may increase the permeability of spores to taurocholate, proper assembly of the appendage is also important for germination. Consistent with an overall enhancement of germination, acdeMmutant shows increased virulence in a hamster model of disease. For a wild type population, spores with a short or no appendage germinate slower than the appendage-bearing spores. Differences in transmission, persistence and disease severity may result, in part, from their proportion in a spore population.
- Published
- 2018
- Full Text
- View/download PDF
3. Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs
- Author
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Jauffret-Roustide, Marie, Chollet, A, Santos, A, Benoit, T, Péchiné, S, Duplessy, C, Bara, J-L, Lévi, Y, Karolak, S, Néfau, T, CERMES3 - Centre de recherche Médecine, sciences, santé, santé mentale, société (CERMES3 - UMR 8211 / U988 / UM 7), and École des hautes études en sciences sociales (EHESS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
paraphernalia ,user acceptability ,bacterial and fungal risk ,injecting practice ,filtering tool ,[SHS]Humanities and Social Sciences - Abstract
International audience; Introduction and Aims : :. People who inject drugs (PWID) are exposed to associated viral, bacterial and fungal risks. These risks can be reduced by filtration. Large disparities in the quality of filtration exist between the various available filters. This paper compares both performance and user acceptability of three filters for drug injection (cotton filters, Sterifilt® and wheel filters) by combining epidemiological and bacteriological analyses. Design and Methods. A cross-sectional epidemiological study (ANRS-Coquelicot) using time–location sampling combined with the generalised weight sampling method was conducted among 985 PWID in France. Two filtration-based bacteriological studies of 0.20- and 0.45-μm wheel filters, Sterifilt filters and cotton filters were also conducted. Results. The bacteriological study highlighted the value of using wheel filters with a porosity of less than 0.5 μm, as they limit the risk of bacterial and fungal infection. The results of this study clearly highlight a distinction between the efficiency of Sterifilt and wheel filters, the latter being more effective. Our epidemiological study highlighted that the use of cotton filters is widespread and routine, but is the subject of much criticism among PWID. Sterifilt is not widely used, and its adoption is slow. Finally, the wheel filter remains a largely untested tool. Discussion and Conclusions. Low product retention and ease of use are the two most important factors for filters for PWID. Bacterial and fungal risk filtration is less important. It is essential to educate PWID about the benefits of wheel filters. [Jauffret-Roustide M, Chollet A, Santos A, Benoit T, Péchiné S, Duplessy C, Bara J-L, Lévi Y, Karolak S, Néfau T. Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs. Drug Alcohol Rev 2017;00:000-000]
- Published
- 2018
4. Protection against Clostridium difficile infection in a hamster model by oral vaccination using flagellin FliC-loaded pectin beads
- Author
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Bruxelle, J.F., primary, Tsapis, N., additional, Hoys, S., additional, Collignon, A., additional, Janoir, C., additional, Fattal, E., additional, and Péchiné, S., additional
- Published
- 2018
- Full Text
- View/download PDF
5. The state of play of rodent models for the study of Clostridioides difficile infection.
- Author
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Brosse A, Coullon H, Janoir C, and Péchiné S
- Subjects
- Animals, Mice, Cricetinae, Humans, Rodentia microbiology, Germ-Free Life, Clostridium Infections microbiology, Disease Models, Animal, Clostridioides difficile pathogenicity
- Abstract
Clostridioides difficile is the most common cause of nosocomial antibiotic-associated diarrhoea and is responsible for a spectrum of diseases characterized by high levels of recurrence and morbidity. In some cases, complications can lead to death. Currently, several types of animal models have been developed to study various aspects of C. difficile infection (CDI), such as colonization, virulence, transmission and recurrence. These models have also been used to test the role of environmental conditions, such as diet, age and microbiome that modulate infection outcome, and to evaluate several therapeutic strategies. Different rodent models have been used successfully, such as the hamster model and the gnotobiotic and conventional mouse models. These models can be applied to study either the initial CDI infectious process or recurrences. The applications of existing rodent models and their advantages and disadvantages are discussed here.
- Published
- 2024
- Full Text
- View/download PDF
6. Anti-S-layer monoclonal antibodies impact Clostridioides difficile physiology.
- Author
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Hunault L, Auria E, England P, Deschamps J, Briandet R, Kremer V, Iannascoli B, Vidal-Maison L, Guo C, Macdonald L, Péchiné S, Denève-Larrazet C, Dupuy B, Gorochov G, Bruhns P, and Sterlin D
- Subjects
- Adult, Humans, Antibodies, Monoclonal therapeutic use, Cell Death, Clostridioides difficile, Gastrointestinal Microbiome
- Abstract
Clostridioides difficile ( C. difficile ), a gram-positive anaerobic and spore-forming bacterium, is the leading cause of nosocomial antibiotic-associated diarrhea in adults which is characterized by high levels of recurrence and mortality. Surface (S)-layer Protein A (SlpA), the most abundantly expressed protein on the bacterial surface, plays a crucial role in the early stages of infection although the nature of its involvement in C. difficile physiology is yet to be fully understood. Anti-S-layer antibodies have been identified in the sera of convalescent patients and have been correlated with improved outcomes of C. difficile infection (CDI). However, the precise mechanisms by which anti-S-layer antibodies confer protection to the host remain unknown. In this study, we report the first monoclonal antibodies (mAbs) targeting the S-layer of reference strain 630. Characterization of these mAbs unraveled important roles for the S-layer protein in growth, toxin secretion, and biofilm formation by C. difficile , with differential and even opposite effects of various anti-SlpA mAbs on these functions. Moreover, one anti-SlpA mAb impaired C. difficile growth and conferred sensitivity to lysozyme-induced lysis. The results of this study show that anti-S-layer antibody responses can be beneficial or harmful for the course of CDI and provide important insights for the development of adequate S-layer-targeting therapeutics.
- Published
- 2024
- Full Text
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7. Immunization Strategies Against Clostridioides difficile.
- Author
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Campidelli C, Bruxelle JF, Collignon A, and Péchiné S
- Subjects
- Animals, Humans, Immunization, Vaccination, Immunization, Passive, Virulence Factors, Clostridioides difficile
- Abstract
Clostridioides difficile (C. difficile) infection (CDI) is an important healthcare but also a community-associated disease. CDI is considered a public health threat and an economic burden. A major problem is the high rate of recurrences. Besides classical antibiotic treatments, new therapeutic strategies are needed to prevent infection, to treat patients, and to prevent recurrences. If fecal transplantation has been recommended to treat recurrences, another key approach is to elicit immunity against C. difficile and its virulence factors. Here, after a summary concerning the virulence factors, the host immune response against C. difficile, and its role in the outcome of disease, we review the different approaches of passive immunotherapies and vaccines developed against CDI. Passive immunization strategies are designed in function of the target antigen, the antibody-based product, and its administration route. Similarly, for active immunization strategies, vaccine antigens can target toxins or surface proteins, and immunization can be performed by parenteral or mucosal routes. For passive immunization and vaccination as well, we first present immunization assays performed in animal models and second in humans and associated clinical trials. The different studies are presented according to the mode of administration either parenteral or mucosal and the target antigens and either toxins or colonization factors., (© 2024. The Author(s), under exclusive license to Springer Nature Switzerland AG.)
- Published
- 2024
- Full Text
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8. Impact of Subinhibitory Concentrations of Metronidazole on Morphology, Motility, Biofilm Formation and Colonization of Clostridioides difficile .
- Author
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Doan TH, Bernet-Camard MF, Hoÿs S, Janoir C, and Péchiné S
- Abstract
Clostridioides difficile infection (CDI) is the primary cause of health-care-associated infectious diarrhea. Treatment requires mostly specific antibiotics such as metronidazole (MTZ), vancomycin or fidaxomicin. However, approximately 20% of treated patients experience recurrences. Treatment with MTZ is complicated by reduced susceptibility to this molecule, which could result in high failure and recurrence rates. However, the mechanism remains unclear. In this study, we investigated the impact of subinhibitory concentrations of MTZ on morphology, motility, biofilm formation, bacterial adherence to the intestinal Caco-2/TC7 differentiated monolayers, and colonization in monoxenic and conventional mouse models of two C. difficile strains (VPI 10463 and CD17-146), showing different susceptibility profiles to MTZ. Our results revealed that in addition to the inhibition of motility and the downregulation of flagellar genes for both strains, sub-inhibitory concentrations of MTZ induced various in vitro phenotypes for the strain CD17-146 exhibiting a reduced susceptibility to this antibiotic: elongated morphology, enhanced biofilm production and increased adherence to Caco-2/TC7 cells. Weak doses of MTZ induced higher level of colonization in the conventional mouse model and a trend to thicker 3-D structures entrapping bacteria in monoxenic mouse model. Thus, sub-inhibitory concentrations of MTZ can have a wide range of physiological effects on bacteria, which may contribute to their persistence after treatment.
- Published
- 2022
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9. Impact of subinhibitory concentrations of metronidazole on proteome of Clostridioides difficile strains with different levels of susceptibility.
- Author
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Doan TH, Yen-Nicolaÿ S, Bernet-Camard MF, Martin-Verstraete I, and Péchiné S
- Subjects
- Adaptation, Physiological, Biofilms drug effects, Biofilms growth & development, Chromatography, Liquid, Clostridioides difficile drug effects, Clostridioides difficile metabolism, Dose-Response Relationship, Drug, Drug Resistance, Bacterial, Gene Expression Regulation, Bacterial drug effects, Sigma Factor metabolism, Tandem Mass Spectrometry, Anti-Bacterial Agents pharmacology, Bacterial Proteins metabolism, Clostridioides difficile growth & development, Metronidazole pharmacology, Proteomics methods
- Abstract
Clostridioides difficile is responsible for various intestinal symptoms from mild diarrhea to severe pseudomembranous colitis and is the primary cause of antibiotic-associated diarrhea in adults. Metronidazole was the first-line treatment for mild to moderate C. difficile infections for 30 years. However, clinical failure and recurrence rates of metronidazole is superior to oral vancomycin and metronidazole is now recommended only as an alternative to vancomycin or fidaxomicin, for an initial non-severe infection. The mechanisms of treatment failure and infection recurrence remain unclear. Given the poor fecal concentrations of metronidazole, the bacteria may be exposed to subinhibitory concentrations of metronidazole and develop adaptation strategy, which is likely to be the origin of an increase in treatment failures. In this study, a proteomic approach was used to analyze changes in the proteome of two strains with different levels of susceptibility to metronidazole in the presence of subinhibitory concentrations of this antibiotic. The two strains were grown to stationary phase: CD17-146, a clinical C. difficile isolate with reduced susceptibility to metronidazole, and VPI 10463, a metronidazole susceptible strain. Our study revealed that, whatever the strain, subinhibitory concentrations of metronidazole modified the amount of proteins involved in protein biosynthesis, glycolysis, and protection against stress induced by metronidazole, as well as in DNA repair. Several proteins involved in stress response are known to be synthesized under the control of Sigma factor B, which suggests a close link between Sigma factor B and metronidazole. Interestingly, impact of metronidazole on protein production for VPI 10463 strain differed from CD17-146 strain, for which the amount of two proteins involved in biofilm formation of CD17-146 were modified by metronidazole., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2020
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10. Molecular features of lipoprotein CD0873: A potential vaccine against the human pathogen Clostridioides difficile .
- Author
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Bradshaw WJ, Bruxelle JF, Kovacs-Simon A, Harmer NJ, Janoir C, Péchiné S, Acharya KR, and Michell SL
- Subjects
- Animals, Colony Count, Microbial, Disease Models, Animal, Female, Humans, Immunization, Immunoglobulin A, Secretory metabolism, Intestines microbiology, Intestines pathology, Ligands, Lipoproteins chemistry, Mice, Inbred C57BL, Mutation genetics, Recombinant Proteins immunology, Bacterial Vaccines immunology, Clostridioides difficile immunology, Clostridium Infections immunology, Clostridium Infections microbiology, Lipoproteins genetics, Lipoproteins immunology
- Abstract
Clostridioides difficile is the primary cause of antibiotic-associated diarrhea and colitis, a healthcare-associated intestinal disease resulting in a significant fatality rate. Colonization of the gut is critical for C. difficile pathogenesis. The bacterial molecules essential for efficient colonization therefore offer great potential as vaccine candidates. Here we present findings demonstrating that the C. difficile immunogenic lipoprotein CD0873 plays a critical role in pathogen success in vivo We found that in a dixenic colonization model, a CD0873-positive strain of C. difficile significantly outcompeted a CD0873-negative strain. Immunization of mice with recombinant CD0873 prevented long-term gut colonization and was correlated with a strong secretory IgA immune response. We further present high-resolution crystal structures of CD0873, at 1.35-2.50 Å resolutions, offering a first view of the ligand-binding pocket of CD0873 and provide evidence that this lipoprotein adhesin is part of a tyrosine import system, an amino acid key in C. difficile infection. These findings suggest that CD0873 could serve as an effective component in a vaccine against C. difficile ., Competing Interests: The authors declare that they have no conflicts of interest with the contents of this article., (© 2019 Bradshaw et al.)
- Published
- 2019
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11. 2,4-Disubstituted 5-Nitroimidazoles Potent against Clostridium difficile.
- Author
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Spitz C, Mathias F, Péchiné S, Doan THD, Innocent J, Pellissier S, Di Giorgio C, Crozet MD, Janoir C, and Vanelle P
- Subjects
- Animals, Anti-Bacterial Agents pharmacology, CHO Cells, Cell Survival drug effects, Clostridioides difficile drug effects, Cricetulus, Drug Design, Drug Resistance, Bacterial drug effects, Humans, Metronidazole pharmacology, Molecular Structure, Nitroimidazoles pharmacology, Structure-Activity Relationship, Sulfones chemistry, Anti-Bacterial Agents chemical synthesis, Clostridium Infections drug therapy, Nitroimidazoles chemical synthesis
- Abstract
Metronidazole is one of the first-line treatments for non-severe Clostridium difficile infections (CDI). However, resistance limits its use in cases of severe and complicated CDI. Structure-activity relationships previously described for the 5-nitroimidazole series have shown that functionalization at the 2- and 4-positions can impart better activity against parasites and anaerobic bacteria than metronidazole. Herein we report the synthesis of new 2,4-disubstituted 5-nitroimidazole compounds that show potent antibacterial activity against C. difficile. We used a vicarious nucleophilic substitution of hydrogen (VNS) reaction to introduce a phenylmethylsulfone at the 4-position and a unimolecular radical nucleophilic substitution (S
RN 1) reaction to introduce an ethylenic function at the 2-position of the 5-nitroimidazole scaffold., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)- Published
- 2019
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12. Prospective evaluation of the adaptive immune response to SlpA in Clostridium difficile infection.
- Author
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Mizrahi A, Bruxelle JF, Péchiné S, and Le Monnier A
- Subjects
- Adaptive Immunity, Antibodies, Bacterial blood, Antibodies, Bacterial immunology, Bacterial Proteins genetics, Clostridioides difficile genetics, Clostridioides difficile isolation & purification, Clostridioides difficile physiology, Clostridium Infections blood, Clostridium Infections immunology, Humans, Immunoglobulin G blood, Immunoglobulin G immunology, Prospective Studies, Bacterial Proteins immunology, Clostridioides difficile immunology, Clostridium Infections microbiology
- Abstract
Clostridium difficile is a major cause of healthcare-associated diarrhea. SlpA is the precursor of the S-layer of C. difficile. The aim of this work was to evaluate the humoral immune response of hospitalized patients to SlpA and its potential role on CDI outcome. Sera of 87 included patients were collected the day of CDI diagnosis as well as at early and late periods after infection. SlpA appeared to be immunogenic with a heterogeneous response between patients after a CDI. Patients with a single episode had a significantly higher anti-SlpA IgG antibody level than patients with recurrent CDI (p = 0.0197). These preliminary results will be useful to understand better the inter-individual variability of immune responses to C. difficile as well as for the development of new therapeutics., (Copyright © 2018 Elsevier Ltd. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
13. Targeting Clostridium difficile Surface Components to Develop Immunotherapeutic Strategies Against Clostridium difficile Infection.
- Author
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Péchiné S, Bruxelle JF, Janoir C, and Collignon A
- Abstract
New therapies are needed to prevent and treat Clostridium difficile infection and to limit the rise in antibiotic resistance. Besides toxins, several surface components have been characterized as colonization factors and have been shown as immunogenic. This review will focus on passive and active immunization strategies targeting C. difficile surface components to combat C. difficile . Concerning passive immunization, the first strategies used antisera raised against the entire bacterium to prevent infection in the hamster model. Then, surface components such as the flagellin and the S-layer proteins were used for immunization and the passive transfer of antibodies was protective in animal models. Passive immunotherapy with polyvalent immunoglobulins was used in humans and bovine immunoglobulin concentrates were evaluated in clinical trials. Concerning active immunization, vaccine assays targeting surface components were tested mainly in animal models, mouse models of colonization and hamster models of infection. Bacterial extracts, spore proteins and surface components of vegetative cells such as cell wall proteins, flagellar proteins, and polysaccharides were used as vaccine targets. Vaccine assays were performed by parenteral and mucosal routes of immunization. Both gave promising results and pave the way to development of new vaccines.
- Published
- 2018
- Full Text
- View/download PDF
14. Understanding Clostridium difficile Colonization.
- Author
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Crobach MJT, Vernon JJ, Loo VG, Kong LY, Péchiné S, Wilcox MH, and Kuijper EJ
- Subjects
- Cross Infection microbiology, Cross Infection transmission, Humans, Risk Factors, Clostridioides difficile growth & development, Clostridium Infections microbiology, Clostridium Infections transmission, Gastrointestinal Tract microbiology
- Abstract
Clostridium difficile is the main causative agent of antibiotic-associated and health care-associated infective diarrhea. Recently, there has been growing interest in alternative sources of C. difficile other than patients with Clostridium difficile infection (CDI) and the hospital environment. Notably, the role of C. difficile -colonized patients as a possible source of transmission has received attention. In this review, we present a comprehensive overview of the current understanding of C. difficile colonization. Findings from gut microbiota studies yield more insights into determinants that are important for acquiring or resisting colonization and progression to CDI. In discussions on the prevalence of C. difficile colonization among populations and its associated risk factors, colonized patients at hospital admission merit more attention, as findings from the literature have pointed to their role in both health care-associated transmission of C. difficile and a higher risk of progression to CDI once admitted. C. difficile colonization among patients at admission may have clinical implications, although further research is needed to identify if interventions are beneficial for preventing transmission or overcoming progression to CDI., (Copyright © 2018 American Society for Microbiology.)
- Published
- 2018
- Full Text
- View/download PDF
15. Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs.
- Author
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Jauffret-Roustide M, Chollet A, Santos A, Benoit T, Péchiné S, Duplessy C, Bara JL, Lévi Y, Karolak S, and Néfau T
- Subjects
- Adult, Female, Filtration, Humans, Male, Middle Aged, Needle Sharing, Drug Users, Harm Reduction, Infection Control methods, Infections etiology, Substance Abuse, Intravenous complications
- Abstract
Introduction and Aims: People who inject drugs (PWID) are exposed to associated viral, bacterial and fungal risks. These risks can be reduced by filtration. Large disparities in the quality of filtration exist between the various available filters. This paper compares both performance and user acceptability of three filters for drug injection (cotton filters, Sterifilt
® and wheel filters) by combining epidemiological and bacteriological analyses., Design and Methods: A cross-sectional epidemiological study (ANRS-Coquelicot) using time-location sampling combined with the generalised weight sampling method was conducted among 985 PWID in France. Two filtration-based bacteriological studies of 0.20- and 0.45-μm wheel filters, Sterifilt filters and cotton filters were also conducted., Results: The bacteriological study highlighted the value of using wheel filters with a porosity of less than 0.5 μm, as they limit the risk of bacterial and fungal infection. The results of this study clearly highlight a distinction between the efficiency of Sterifilt and wheel filters, the latter being more effective. Our epidemiological study highlighted that the use of cotton filters is widespread and routine, but is the subject of much criticism among PWID. Sterifilt is not widely used, and its adoption is slow. Finally, the wheel filter remains a largely untested tool., Discussion and Conclusions: Low product retention and ease of use are the two most important factors for filters for PWID. Bacterial and fungal risk filtration is less important. It is essential to educate PWID about the benefits of wheel filters. [Jauffret-Roustide M, Chollet A, Santos A, Benoit T, Péchiné S, Duplessy C, Bara J-L, Lévi Y, Karolak S, Néfau T. Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs. Drug Alcohol Rev 2018;37:106-115]., (© 2017 Australasian Professional Society on Alcohol and other Drugs.)- Published
- 2018
- Full Text
- View/download PDF
16. Immunization Strategies Against Clostridium difficile.
- Author
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Bruxelle JF, Péchiné S, and Collignon A
- Subjects
- Animals, Antibodies, Bacterial immunology, Bacterial Vaccines immunology, Clostridioides difficile pathogenicity, Host-Pathogen Interactions immunology, Humans, Clostridium Infections immunology, Immunization
- Abstract
C. difficile infection (CDI) is an important healthcare- but also community-associated disease. CDI is considered a public health threat and an economic burden. A major problem is the high rate of recurrences. Besides classical antibiotic treatments, new therapeutic strategies are needed to prevent infection, to treat patients and prevent recurrences. If fecal transplantation has been recommended to treat recurrences, another key approach is to restore immunity against C. difficile and its virulence factors. Here, after a summary concerning the virulence factors, the host immune response against C. difficile and its role in the outcome of disease, we review the different approaches of passive immunotherapies and vaccines developed against CDI. Passive immunization strategies are designed in function of the target antigen, the antibody-based product and its administration route. Similarly, for active immunization strategies, vaccine antigens can target toxins or surface proteins and immunization can be performed by parenteral or mucosal routes. For passive immunization and vaccination as well, we first present immunization assays performed in animal models and second in humans and associated clinical trials. The different studies are presented according to the mode of administration either parenteral or mucosal and the target antigens, either toxins or colonization factors.
- Published
- 2018
- Full Text
- View/download PDF
17. Clostridium difficile flagellin FliC: Evaluation as adjuvant and use in a mucosal vaccine against Clostridium difficile.
- Author
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Bruxelle JF, Mizrahi A, Hoÿs S, Collignon A, Janoir C, and Péchiné S
- Subjects
- Animals, Antibodies, Bacterial immunology, Cell Count, Clostridioides difficile drug effects, Clostridioides difficile metabolism, Colony Count, Microbial, Enterocolitis, Pseudomembranous blood, Enterocolitis, Pseudomembranous immunology, Enterocolitis, Pseudomembranous microbiology, Female, Immunity drug effects, Immunization, Immunoglobulin G blood, Intestinal Mucosa drug effects, Intestinal Mucosa immunology, Intestinal Mucosa microbiology, Membrane Glycoproteins immunology, Mice, Inbred C57BL, Ovalbumin immunology, Rectum immunology, Vaccination, Adjuvants, Immunologic pharmacology, Bacterial Proteins immunology, Bacterial Vaccines immunology, Clostridioides difficile immunology, Flagellin metabolism, Immunity, Mucosal drug effects
- Abstract
The immunogenicity of bacterial flagellin has been reported in different studies. By its close interaction with the immune system, the flagellin represents an interesting adjuvant and vaccine candidate. Salmonella Typhimurium flagellin has already been tested as adjuvant to stimulate mucosal immunity. Here, we assessed the ability of Clostridium difficile flagellin FliC to act as a mucosal adjuvant, first combined with ovalbumin as antigen and second with a C. difficile surface protein, the precursor of the S-layer proteins SlpA. Using ovalbumin as antigen, we compared the gut mucosal adjuvanticity of FliC to Salmonella Typhimurium flagellin and cholera toxin. Two routes of immunization were tested in a mouse model: intra-rectal and intra-peritoneal, following which, gut mucosal and systemic antibody responses against ovalbumin (Immunoglobulins G and Immunoglobulins A) were analyzed by Enzyme-Linked Immuno Assay in intestinal contents and in sera. In addition, ovalbumin-specific immunoglobulin producing cells were detected in the intestinal lamina propria by Enzyme-Linked Immunospot. Results showed that FliC as adjuvant for immunization targeting ovalbumin was able to stimulate a gut mucosal and systemic antibody response independently of the immunization route. In order to develop a mucosal vaccine to prevent C. difficile intestinal colonization, we assessed in a mouse model the efficacy of FliC as adjuvant compared with cholera toxin co-administrated with the C. difficile S-layer precursor SlpA as antigen. After challenge, a significant decrease of C. difficile intestinal colonization was observed in immunized groups compared to the control group. Our results showed that C. difficile FliC could be used as adjuvant in mucosal vaccination strategy against C. difficile infections.
- Published
- 2017
- Full Text
- View/download PDF
18. Emerging monoclonal antibodies against Clostridium difficile infection.
- Author
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Péchiné S, Janoir C, and Collignon A
- Subjects
- Animals, Antibodies, Monoclonal immunology, Bacterial Proteins administration & dosage, Bacterial Proteins immunology, Clostridium Infections diagnosis, Clostridium Infections immunology, Disease Models, Animal, Drug Delivery Systems trends, Humans, Vaccines administration & dosage, Vaccines immunology, Antibodies, Monoclonal administration & dosage, Clostridioides difficile, Clostridium Infections drug therapy, Immunization, Passive methods
- Abstract
Introduction: Clostridium difficile infections are characterized by a high recurrence rate despite antibiotic treatments and there is an urgent need to develop new treatments such as fecal transplantation and immonotherapy. Besides active immunotherapy with vaccines, passive immunotherapy has shown promise, especially with monoclonal antibodies. Areas covered: Herein, the authors review the different assays performed with monoclonal antibodies against C. difficile toxins and surface proteins to treat or prevent primary or recurrent episodes of C. difficile infection in animal models and in clinical trials as well. Notably, the authors lay emphasis on the phase III clinical trial (MODIFY II), which allowed bezlotoxumab to be approved by the Food and Drug Administration and the European Medicines Agency. They also review new strategies for producing single domain antibodies and nanobodies against C. difficile and new approaches to deliver them in the digestive tract. Expert opinion: Only two human Mabs against TcdA and TcdB have been tested alone or in combination in clinical trials. However, many animal model studies have provided rationale for the use of Mabs and nanobodies in C. difficile infection and pave the way for further clinical investigation.
- Published
- 2017
- Full Text
- View/download PDF
19. Immune responses induced by Clostridium difficile.
- Author
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Péchiné S and Collignon A
- Subjects
- Adaptive Immunity, Animals, Antibodies, Bacterial physiology, Bacterial Proteins immunology, Bacterial Toxins immunology, Enterocolitis, Pseudomembranous microbiology, Humans, Immunity, Innate, Intestinal Mucosa immunology, Intestinal Mucosa microbiology, Virulence Factors immunology, Clostridioides difficile immunology, Enterocolitis, Pseudomembranous immunology
- Abstract
The spectrum of Clostridium difficile infections is highly variable, ranging from asymptomatic carriage to fatal colitis depending on the strain virulence and on the host, its gut microbiota and its immune response. After disruption of the gut microbiota, C. difficile pathogenesis can be divided into three steps: 1) contamination by spores and their germination; 2) multiplication of vegetative cells and intestinal colonization using colonization factors; 3) production of the toxins TcdA and TcdB, and for some strains, the binary toxin, which are responsible for the clinical signs. Three lines of defense counteract C. difficile. The first line is the epithelial barrier, which is breached by the toxins. Then, a rapid innate immune response follows, which forms the second line of defense. It provides very quick defense reactions against C. difficile but is non-specific and does not confer memory. C. difficile and its virulence factors, the toxins and colonization factors, induce a highly pro-inflammatory response, which can be either beneficial or harmful, but triggers the adaptive immunity as the third line of defense required to control the infectious process. Adaptive immunity provides a highly specific immune response against C. difficile with memory and long lasting immunity. The innate and adaptive immune responses against the toxins and surface components are analyzed as well as their role in disease susceptibility, severity and recurrences., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
20. Clostridium difficile Adhesins.
- Author
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Péchiné S, Denève-Larrazet C, and Collignon A
- Subjects
- ADP Ribose Transferases genetics, ADP Ribose Transferases metabolism, Adhesins, Bacterial metabolism, Animals, Bacterial Adhesion, Bacterial Proteins genetics, Bacterial Proteins metabolism, Bacterial Toxins genetics, Bacterial Toxins metabolism, Caco-2 Cells, Chaperonin 60 genetics, Chaperonin 60 metabolism, Chlorocebus aethiops, Clostridioides difficile metabolism, Clostridium Infections microbiology, Clostridium Infections pathology, Enterotoxins genetics, Enterotoxins metabolism, Fibronectins chemistry, Fibronectins metabolism, Germ-Free Life, Humans, Intestines microbiology, Intestines pathology, Mice, Microscopy, Immunoelectron methods, Periplasmic Binding Proteins metabolism, Protein Binding, Vero Cells, Virulence, Adhesins, Bacterial genetics, Clostridioides difficile genetics, Clostridioides difficile pathogenicity, Gene Expression Regulation, Bacterial, Periplasmic Binding Proteins genetics
- Abstract
Clostridium difficile is responsible for a large spectrum of intestinal diseases ranging from mild diarrhea to fatal colitis depending on the one hand on the strain virulence and on the other on the host. The pathogenesis of C. difficile infection could be seen as a three-step process that takes place after disruption of the digestive microbiota by antibiotics: (1) contamination by and germination of spores; (2) multiplication of vegetative cells in the colonic niche using colonization factors; (3) production of the two toxins TcdA and TcdB and for some strains an additional toxin, the binary toxin CDT. Several studies have been performed to characterize the bacterial factors involved in the colonization step and particularly adhesins.Here, we describe first the methods used to study C. difficile adherence in vitro to epithelial cells and in vivo in animal model intestinal tract, and second the methods used to demonstrate the adhesive properties of surface proteins using Cwp66, GroEL, and FbpA as examples.
- Published
- 2016
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21. Passive and active immunization strategies against Clostridium difficile infections: state of the art.
- Author
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Mizrahi A, Collignon A, and Péchiné S
- Subjects
- Animals, Clinical Trials as Topic, Clostridium Infections immunology, Drug Evaluation, Preclinical, Humans, Clostridioides difficile immunology, Clostridium Infections prevention & control, Immunization, Passive methods, Vaccination methods
- Abstract
The pathophysiology of Clostridium difficile infections (CDI) could be considered as a three-step process that takes place after disruption of the digestive microbiota by antibiotics: 1) germination of spores; 2) multiplication and persistence of C. difficile in the colonic niche thanks to colonization factors; 3) production of the two toxins TcdA and TcdB and for some strains an additional toxin, the binary toxin CDT. Different immunization strategies against C. difficile have been developed, first regarding the toxins. Immunization assays with colonization factors have followed, and allowed accumulation of new data concerning theirs functions and immunogenicity. Here, we present the toxins, the colonization factors and their use in passive and active immunizations to treat and/or to prevent C. difficile infections. The various experiments performed in animal models and the first clinical trials in humans are reported., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
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- View/download PDF
22. Immunization using GroEL decreases Clostridium difficile intestinal colonization.
- Author
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Péchiné S, Hennequin C, Boursier C, Hoys S, and Collignon A
- Subjects
- Animals, Antibodies, Bacterial blood, Antibodies, Bacterial immunology, Antibody Specificity immunology, Bacterial Proteins metabolism, Bacterial Vaccines immunology, Cell Wall immunology, Cell Wall metabolism, Chaperonin 60 metabolism, Clostridioides difficile metabolism, Cricetinae, Disease Models, Animal, Enterocolitis, Pseudomembranous immunology, Enterocolitis, Pseudomembranous microbiology, Enterocolitis, Pseudomembranous mortality, Female, Immunization, Mice, Proteomics methods, Recombinant Proteins immunology, Bacterial Proteins immunology, Chaperonin 60 immunology, Clostridioides difficile immunology, Intestines immunology, Intestines microbiology
- Abstract
Clostridium difficile is a pathogen which is responsible for diarrhea and colitis, particularly after treatment with antibiotics. Clinical signs are mainly due to two toxins, TcdA and TcdB. However, the first step of pathogenesis is the colonization process. We evaluated C. difficile surface proteins as vaccine antigens in the hamster model to prevent intestinal colonization. This vaccination induced a partial protection of hamsters against death after a C. difficile challenge. A proteomic analysis of animal sera allowed us to identify proteins which could be responsible for the protection observed. Among these proteins, we identified the GroEL heat shock protein. To confirm the role of the specific GroEL antibodies in the delayed C. difficile colonization of hamsters, we performed an immunization assay in a mouse model. After intranasal immunization with the recombinant protein GroEL, we observed a lower C. difficile intestinal colonization in the immunized group as compared to the control group.
- Published
- 2013
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23. Encapsulation of Cwp84 into pectin beads for oral vaccination against Clostridium difficile.
- Author
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Sandolo C, Péchiné S, Le Monnier A, Hoys S, Janoir C, Coviello T, Alhaique F, Collignon A, Fattal E, and Tsapis N
- Subjects
- Administration, Oral, Animals, Anti-Bacterial Agents administration & dosage, Anti-Bacterial Agents therapeutic use, Antibodies, Bacterial blood, Bacterial Vaccines administration & dosage, Chemistry, Pharmaceutical, Clostridioides difficile enzymology, Clostridium Infections immunology, Cricetinae, Cysteine Endopeptidases administration & dosage, Cysteine Endopeptidases chemistry, Disease Models, Animal, Drug Compounding, Drug Stability, Enzyme-Linked Immunosorbent Assay, Female, Immunoglobulin G blood, Molecular Weight, Survival Analysis, Bacterial Vaccines immunology, Clostridioides difficile immunology, Clostridium Infections prevention & control, Cysteine Endopeptidases immunology, Drug Carriers chemistry, Pectins chemistry
- Abstract
We have designed an oral vaccine against Clostridium difficile infection. The virulent factor Cwp84, that is a cystein protease highly immunogenic in patients with C. difficile-associated disease, was entrapped within pectin beads. Beads encapsulating Cwp84 were shown to be stable in the simulated intestinal medium and to release the cystein protease once in the simulated colonic medium. Three groups of hamsters were immunized, the first receiving pectin beads encapsulating Cwp84, the second unloaded beads and the third one free Cwp84. After three immunizations by the intragastric route, all groups received clindamycine. Post-challenge survival with a strain of C. difficile showed that 2 days after infection, all hamsters treated with unloaded beads and all hamsters treated with free Cwp84 have deceased after 7 days, whereas about 40% of hamsters administered with Cwp84-loaded beads survived 10 days after challenge, proving that oral vaccination provides partial protection. These first data obtained with an oral vaccine against C. difficile appear promising for preventing this infection., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
24. Immunization of hamsters against Clostridium difficile infection using the Cwp84 protease as an antigen.
- Author
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Péchiné S, Denève C, Le Monnier A, Hoys S, Janoir C, and Collignon A
- Subjects
- Administration, Rectal, Animals, Antibodies, Bacterial blood, Antigens, Bacterial immunology, Bacterial Vaccines administration & dosage, Clostridioides difficile enzymology, Clostridium Infections immunology, Cricetinae, Disease Models, Animal, Gastric Lavage, Gastrointestinal Tract microbiology, Injections, Subcutaneous, Male, Survival Analysis, Bacterial Vaccines immunology, Clostridioides difficile immunology, Clostridium Infections prevention & control, Cysteine Endopeptidases immunology, Immunization methods
- Abstract
Clostridium difficile is a pathogen responsible for diarrhoea and colitis, particularly after antibiotic treatment. We evaluated the C. difficile protease Cwp84, found to be associated with the S-layer proteins, as a vaccine antigen to limit the C. difficile intestinal colonization and therefore the development of the infection in a clindamycin-treated hamster model. First, we evaluated the immune response and the animal protection against death induced by several immunization routes: rectal, intragastric and subcutaneous. Antibody production was variable according to the immunization routes. In addition, serum Cwp84 antibody titres did not always correlate with animal protection after challenge with a toxigenic C. difficile strain. The best survival rate was observed with the rectal route of immunization. Then, in a second assay, we selected this immunization route to perform a larger immunization assay including a Cwp84 immunized group and a control group. Clostridium difficile intestinal colonization and survival rate, as well as the immune response were examined. Clostridium difficile hamster challenge resulted in a 26% weaker and slower C. difficile intestinal colonization in the immunized group. Furthermore, hamster survival in the Cwp84 immunized group was 33% greater than that of the control group, with a significant statistical difference., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)
- Published
- 2011
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25. Cwp84, a surface-associated protein of Clostridium difficile, is a cysteine protease with degrading activity on extracellular matrix proteins.
- Author
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Janoir C, Péchiné S, Grosdidier C, and Collignon A
- Subjects
- Antibodies, Bacterial metabolism, Bacterial Proteins chemistry, Bacterial Proteins genetics, Caseins metabolism, Cysteine Endopeptidases chemistry, Cysteine Endopeptidases genetics, Cysteine Proteinase Inhibitors pharmacology, Enzyme Stability, Hydrogen-Ion Concentration, Immunoblotting, Membrane Glycoproteins metabolism, Molecular Weight, Recombinant Proteins immunology, Recombinant Proteins isolation & purification, Bacterial Proteins metabolism, Clostridioides difficile enzymology, Cysteine Endopeptidases metabolism, Extracellular Matrix Proteins metabolism
- Abstract
Clostridium difficile pathogenicity is mediated mainly by its A and B toxins, but the colonization process is thought to be a necessary preliminary step in the course of infection. The aim of this study was to characterize the Cwp84 protease of C. difficile, which is highly immunogenic in patients with C. difficile-associated disease and is potentially involved in the pathogenic process. Cwp84 was purified as a recombinant His-tagged protein, and specific antibodies were generated in rabbits. Treatment of multiple-band-containing eluted fractions with a reducing agent or with trypsin led to accumulation of a unique protein species with an estimated molecular mass of 61 kDa, corresponding most likely to mature autoprocessed Cwp84 (mCwp84). mCwp84 showed concentration-dependent caseinolytic activity, with maximum activity at pH 7.5. The Cwp84 activity was inhibited by various cysteine protease inhibitors, such as the specific inhibitor E64, and the anti-Cwp84-specific antibodies. Using fractionation experiments followed by immunoblot detection, the protease was found to be associated with the S-layer proteins, mostly as a nonmature species. Proteolytic assays were performed with extracellular matrix proteins to assess the putative role of Cwp84 in the pathogenicity of C. difficile. No degrading activity was detected with type IV collagen. In contrast, Cwp84 exhibited degrading activity with fibronectin, laminin, and vitronectin, which was neutralized by the E64 inhibitor and specific antibodies. In vivo, this proteolytic activity could contribute to the degradation of the host tissue integrity and to the dissemination of the infection.
- Published
- 2007
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26. Diminished intestinal colonization by Clostridium difficile and immune response in mice after mucosal immunization with surface proteins of Clostridium difficile.
- Author
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Péchiné S, Janoir C, Boureau H, Gleizes A, Tsapis N, Hoys S, Fattal E, and Collignon A
- Subjects
- Administration, Rectal, Animals, Antibodies, Bacterial biosynthesis, Antibodies, Bacterial immunology, Bacterial Outer Membrane Proteins chemistry, Bacterial Proteins chemistry, Bacterial Proteins immunology, Bacterial Vaccines immunology, Cholera Toxin immunology, Cholera Toxin pharmacology, Clostridioides difficile immunology, Cysteine Endopeptidases chemistry, Cysteine Endopeptidases immunology, Female, Humans, Immunity, Mucosal immunology, Immunoglobulin A biosynthesis, Immunoglobulin A immunology, Intestines immunology, Mice, Mice, Inbred C3H, Bacterial Outer Membrane Proteins immunology, Bacterial Vaccines administration & dosage, Clostridioides difficile growth & development, Enterocolitis, Pseudomembranous immunology, Enterocolitis, Pseudomembranous microbiology, Intestines microbiology
- Abstract
Clostridium difficile pathogenesis is mainly due to toxins A and B. However, the first step of pathogenesis is the colonization process. We evaluated C. difficile surface proteins as vaccine antigens to diminish intestinal colonization in a human flora-associated mouse model. First, we used the flagellar cap protein FliD of C. difficile, in order to test several immunization routes: intranasal, rectal, and intragastric. The rectal route, which is the most efficient, was used to vaccine groups of mice with different antigen combinations. After immunizations, the mice were challenged with the toxigenic C. difficile and a significant statistical difference between the control group and the immunized groups was observed in the colonization levels of C. difficile.
- Published
- 2007
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- View/download PDF
27. Variability of Clostridium difficile surface proteins and specific serum antibody response in patients with Clostridium difficile-associated disease.
- Author
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Péchiné S, Janoir C, and Collignon A
- Subjects
- Animals, Bacterial Proteins genetics, Bacterial Proteins immunology, Bacterial Proteins metabolism, Clostridioides difficile genetics, Clostridioides difficile immunology, Enterocolitis, Pseudomembranous microbiology, Flagellin genetics, Flagellin immunology, Flagellin metabolism, Genotype, Humans, Membrane Proteins genetics, Membrane Proteins metabolism, Mice, Phenotype, Polymerase Chain Reaction, Polymorphism, Restriction Fragment Length, Ribotyping, Sensitivity and Specificity, Antibodies, Bacterial blood, Antibody Specificity, Clostridioides difficile classification, Enterocolitis, Pseudomembranous immunology, Genetic Variation, Membrane Proteins immunology
- Abstract
Pathogen attachment is a crucial early step in mucosal infections. This step is mediated by important virulence factors, such as surface proteins. Clostridium difficile surface proteins have been identified as (i) adhesins (the flagellar cap protein FliD; the flagellin FliC; and the cell wall protein Cwp 66 with a two domain-structure [Cw 66 N-terminal and Cwp 66 C-terminal domains]) and (ii) protease (the Cwp 84 protein). To address the roles of these proteins in the pathogenesis of Clostridium difficile and to identify vaccine antigen candidates, we analyzed the variability of the proteins and their immunogenicities in 17 patients with C. difficile-associated disease. PCR-restriction fragment length polymorphism analysis of amplified gene products revealed interstrain homogeneity with fliC and fliD, in contrast to cwp 66 genes. Immunoblot analysis showed that FliC and FliD were detected in the majority of isolates. The N-terminal domain of Cwp 66 and Cwp 84 were present in all strains tested, in contrast to the Cwp 66 C-terminal domain, the expression of which was heterogeneous. The 17 sera from the corresponding patients were analyzed by enzyme-linked immunosorbent assay to detect antibodies directed against these proteins. Many patients developed antibodies to FliC, FliD, Cwp 84, and the Cwp 66 C-terminal domain, but not to the Cwp 66 N-terminal domain. In conclusion, this study confirms the expression of these surface proteins of C. difficile during the course of the disease. In addition, the FliC, FliD, and Cwp 84 proteins appeared to be good potential vaccine candidates.
- Published
- 2005
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28. Immunological properties of surface proteins of Clostridium difficile.
- Author
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Péchiné S, Gleizes A, Janoir C, Gorges-Kergot R, Barc MC, Delmée M, and Collignon A
- Subjects
- Adhesins, Bacterial analysis, Adhesins, Bacterial immunology, Adolescent, Adult, Aged, Bacterial Adhesion, Child, Clostridioides difficile chemistry, Clostridium Infections microbiology, Enzyme-Linked Immunosorbent Assay, Female, Flagellin immunology, Humans, Male, Middle Aged, Antibodies, Bacterial blood, Clostridioides difficile immunology, Clostridium Infections immunology, Membrane Glycoproteins immunology
- Abstract
Sera from patients with Clostridium difficile-associated disease (CDAD) and sera from a control group were analysed by an ELISA to detect antibodies directed against four surface proteins and toxins A and B of C. difficile. The surface proteins were the flagellar cap protein FliD, the flagellin FliC, the adhesin Cwp66 divided into two domains, Cwp66-Nterminal and Cwp66-Cterminal, and the fibronectin-binding protein Fbp68. For each antigen, antibody levels in the CDAD patient group and in the control group were compared. In the CDAD patient group, the mean of the antibody levels decreased from Cwp66-Cterminal to Fbp68, FliD, toxins B and A, Cwp66-Nterminal and finally FliC, suggesting different immunogenic properties among these adhesins. For Cwp66-Nterminal, FliC, FliD and Fbp68, the antibody level observed in the control group was higher than in the CDAD group with a statistically significant difference whereas the antibody level for toxins A and B was not statistically different. In conclusion, this study suggests that during the clinical course of disease, C. difficile adhesins are able to induce an immune response which could play a role in the defence mechanism of the host.
- Published
- 2005
- Full Text
- View/download PDF
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