Page Banner

United States Department of Agriculture

Agricultural Research Service

Title: Identification of Brachyspira (Serpulina) Pilosicoli Membrane Proteins by Use of Monoclonal Antibodies

Authors
item Trott, Darren
item Zuerner, Richard
item Wannemuehler, Mike - VMRI, ISU, AMES, IA.
item Stanton, Thaddeus

Submitted to: Gordon Research Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: January 20, 2000
Publication Date: N/A

Technical Abstract: Brachyspira pilosicoli is the agent of porcine intestinal spirochetosis. The organism also colonizes a range of animal species including humans. A unique feature associated with infection with B. pilosicoli is the attachment of large numbers of spirochetes by one end to the colonic epithelium. The major objective of this study was to identify unique B. pilosicoli outer membrane proteins (OMPs) that may be involved in the host parasite relationship. Membrane vesicles were obtained from B. pilosicoli strain 95-1000 by osmotic lysis and centrifugation. The vesicles were separated by density into four fractions: high density membrane bands A (1.18 g/cm3) and B (1.16 g/cm3) and low density membrane bands A (1.14 g/cm3) and B (1.12 g/cm3). The application of cellular markers showed that each band contained outer membrane and was free of cytoplasmic and flagella contamination. However inner membrane contamination could not be completely yeliminated. Low density membrane band B contained the lowest amount of penicillin binding proteins and was therefore selected as antigen for the production of monoclonal antibodies (MAbs). Six MAbs reacting with B. pilosicoli proteins of 23, 23/24 (doublet), 35, 51, 61 and 79 KDa were produced. The proteins were not present in whole cell lysates of the type strains of B. hyodysenteriae, B. innocens, B. intermedia, B. alvinipulli or B. murdochii. Immunogold labeling with concentrated MAb directed against the 23 KDa protein confirmed that the protein was located on the outer membrane. The MAbs will be useful reagents for identifying unique B. pilosicoli membane proteins and for elucidating the role of OMPs in interactions between B. pilosicoli cells and the host.

Last Modified: 12/21/2014
Footer Content Back to Top of Page