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PMID: 10388671 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Production of monoclonal antibodies to Listeria monocytogenes and their application to determine the virulence of isolates from channel catfish.

Applied and environmental microbiology ·Vol. 65 ·No. 7 ·1999-07-00 ·Pages 2827-32

Erdenlig S, Ainsworth AJ, Austin FW

Abstract

We produced monoclonal antibodies (MAbs) to the extracellular proteins of Listeria monocytogenes EGD grown in Chelex-treated improved minimal medium. Ten of the positive hybridomas generated were chosen for further characterization. Seven of the MAbs reacted with a protein having a molecular mass of 60 kDa. These MAbs inhibited listeriolysin (LLO)-mediated hemolysis, and two of them were specific for LLO and none of the other thiol-activated toxins tested. In an enzyme-linked immunosorbent assay and Western blot analysis, five of the anti-LLO MAbs reacted with ivanolysin from Listeria ivanovii. Three of the 10 MAbs reacted with a 29-kDa protein on Western blots and neutralized the phosphatidylcholine-specific phospholipase C (PC-PLC) activity of L. monocytogenes. These three anti-PC-PLC MAbs did not react with phospholipases from five different gram-positive bacteria. However, the anti-PC-PLC MAbs recognized a 27-kDa extracellular protein from L. ivanovii and neutralized sphingomyelinase activity in a hemolysis test that demonstrates the antigenic relatedness of listerial phospholipases. These data indicate that listerial thiol-activated toxins possess species-specific epitopes and share group-specific epitopes. This is the first description of MAbs that neutralize listerial PC-PLC, and the data suggest that there is antigenic similarity between L. monocytogenes PC-PLC and L. ivanovii sphingomyelinase. The reactions of the MAbs with catfish isolates of L. monocytogenes suggested that some of the isolates examined lack the LLO and/or PC-PLC required for pathogenicity. The MAbs described here differentiated some catfish isolates from previously described type strain-pathogenic isolates and could be useful for detecting and determining the virulence of L. monocytogenes in food and clinical samples and for detecting L. ivanovii in veterinary clinical samples.

MeSH Terms
Animals Antibodies, Bacterial/biosynthesis Antibodies, Monoclonal/biosynthesis,immunology Bacterial Toxins Blotting, Western Enzyme-Linked Immunosorbent Assay Heat-Shock Proteins/immunology Hemolysin Proteins/immunology Hemolysis Ictaluridae/microbiology Listeria monocytogenes/immunology,isolation & purification,pathogenicity Phospholipases/metabolism Toxins, Biological Type C Phospholipases/immunology,metabolism Virulence
Chemicals
Antibodies, Bacterial Antibodies, Monoclonal Bacterial Toxins Heat-Shock Proteins Hemolysin Proteins Toxins, Biological Phospholipases Type C Phospholipases phosphatidylcholine-specific phospholipase C hlyA protein, Listeria monocytogenes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Erdenlig S
Veterinary Medical Research Program, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi 39762, USA.
Ainsworth A J
Austin F W
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.65.7.2827-2832.1999
Published
1999-07-00
Pages
2827-32
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC91424
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.65.7.2827-2832.1999 PMC Full Text View journal details
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