Interrelationships between digestive proteolytic activities and production and quantitation of toxins in pseudomembranous colitis induced by Clostridium difficile in gnotobiotic mice
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| Published in | Infection and Immunity Vol. 57; no. 12; pp. 3922 - 3927 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
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Washington, DC
American Society for Microbiology
01.12.1989
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| AbstractList | Clostridium difficile pathogenicity is related to in vivo production of toxins, and it is of great interest to detect toxins produced in biological samples. Several reports have shown that proteases in stools interfere with immunological methods for quantitation of toxin A. The purpose of this work was to estimate the relationship between the proteases and the C. difficile toxins produced in a gnotobiotic mouse model of pseudomembranous cecitis. Cecal proteolytic activities hydrolyzed toxin A, and immunoglobulin G bound to the microtiter plate used in immunoassays. This interference could be blocked by the addition of trypsin inhibitor to the samples. The ability of soluble toxin A to bind to bound antibodies in an enzyme-linked immunosorbent assay was not affected by the proteases, but the biological activity was reduced 100-fold. The cytotoxicity of toxin B was not modified by proteolytic activity treatment. Mice inoculated with a low toxin A-producing strain of C. difficle did not die, and no modulation of proteolytic activities occurred. Classifications Services IAI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue IAI About IAI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy Connect to IAI IAI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0019-9567 Online ISSN: 1098-5522 Copyright © 2014 by the American Society for Microbiology. For an alternate route to IAI .asm.org, visit: IAI Clostridium difficile pathogenicity is related to in vivo production of toxins, and it is of great interest to detect toxins produced in biological samples. Several reports have shown that proteases in stools interfere with immunological methods for quantitation of toxin A. The purpose of this work was to estimate the relationship between the proteases and the C. difficile toxins produced in a gnotobiotic mouse model of pseudomembranous cecitis. Cecal proteolytic activities hydrolyzed toxin A, and immunoglobulin G bound to the microtiter plate used in immunoassays. This interference could be blocked by the addition of trypsin inhibitor to the samples. The ability of soluble toxin A to bind to bound antibodies in an enzyme-linked immunosorbent assay was not affected by the proteases, but the biological activity was reduced 100-fold. The cytotoxicity of toxin B was not modified by proteolytic activity treatment. Mice inoculated with a low toxin A-producing strain of C. difficile did not died, and no modulation of proteolytic activities occurred. After inoculation with the lethal VPI strain of C. difficile, toxins A and B reached maximum levels in the ceca at 12 h postinfection. At this time, the proteolytic activities did not decrease from the levels seen at zero time. Mice died within 2 days. At this time (about 32 postinfection), proteolytic activities were sharply decreased in the lower parts of the digestive tracts. The findings that serum inhibited the proteases and that there was a 100-fold increase in serum-derived mouse immunoglobulins in the lumen as the C. difficile infection progressed suggest that the decrease in protease activity in the lower digestive tract may be related to the exudation of serum from the inflammation process. |
| Author | G W Elmer M C Muller F Lucas F Dubos-Ramaré G Corthier |
| AuthorAffiliation | Laboratoire d'Ecologie Microbienne, Institut National de la Recherche Agronomique, CRJ, Jouy-en-Josas, France |
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| Keywords | Gnotobiotic animal Digestive system Enzyme Proteinase Rodentia Clostridium difficile Clostridiales Enzyme inhibitor Infection Toxin Vertebrata Mammalia Enzymatic activity Mouse Clostridiaceae Bacteria Pseudomembranous colitis Quantitative analysis |
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| SubjectTerms | Animals Bacterial Proteins Bacterial Toxins - metabolism Bacteriology Biological and medical sciences Clostridium - pathogenicity Digestive System - enzymology Digestive System - metabolism Enterocolitis, Pseudomembranous - microbiology Enterocolitis, Pseudomembranous - physiopathology Enterotoxins - metabolism Fundamental and applied biological sciences. Psychology Germ-Free Life Mice Mice, Inbred C3H Microbiology Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains Peptide Hydrolases - metabolism |
| Title | Interrelationships between digestive proteolytic activities and production and quantitation of toxins in pseudomembranous colitis induced by Clostridium difficile in gnotobiotic mice |
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