Characterization of monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5

Toyotsugu Nakai From Research Center for Veterinary Science of The Kitasato Institute, 11391, Matsugasaki, Kashiwa, Chiba 277 (Nakai, Horiguchi, Kume), and Department of Bacteriology, The Kitasato Institute, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan (Kawahara, Danbara).

Search for other papers by Toyotsugu Nakai in
Current site
Google Scholar
PubMed
Close
 PhD, DVM
,
Kazuyoshi Kawahara From Research Center for Veterinary Science of The Kitasato Institute, 11391, Matsugasaki, Kashiwa, Chiba 277 (Nakai, Horiguchi, Kume), and Department of Bacteriology, The Kitasato Institute, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan (Kawahara, Danbara).

Search for other papers by Kazuyoshi Kawahara in
Current site
Google Scholar
PubMed
Close
 PhD
,
Yasuhiko Horiguchi From Research Center for Veterinary Science of The Kitasato Institute, 11391, Matsugasaki, Kashiwa, Chiba 277 (Nakai, Horiguchi, Kume), and Department of Bacteriology, The Kitasato Institute, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan (Kawahara, Danbara).

Search for other papers by Yasuhiko Horiguchi in
Current site
Google Scholar
PubMed
Close
 PhD, DVM
,
Hirofumi Danbara From Research Center for Veterinary Science of The Kitasato Institute, 11391, Matsugasaki, Kashiwa, Chiba 277 (Nakai, Horiguchi, Kume), and Department of Bacteriology, The Kitasato Institute, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan (Kawahara, Danbara).

Search for other papers by Hirofumi Danbara in
Current site
Google Scholar
PubMed
Close
 PhD
, and
Katsumi Kume From Research Center for Veterinary Science of The Kitasato Institute, 11391, Matsugasaki, Kashiwa, Chiba 277 (Nakai, Horiguchi, Kume), and Department of Bacteriology, The Kitasato Institute, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan (Kawahara, Danbara).

Search for other papers by Katsumi Kume in
Current site
Google Scholar
PubMed
Close
 PhD, DVM

SUMMARY

Two monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5, designated as 5MAb-l and 5MAb-6, were characterized. Enzyme-linked immunosorbent assay-inhibition tests with whole-cell antigens obtained from strains of serotype 1 through 12 of A pleuropneumoniae revealed that 5 MAb-1 bound to only serotype-5 strains. The epitope recognized by 5MAb-l was a carbohydrate that was sensitive to periodate oxidation and resided on the structure of β-l,6-linked d-galactose in an O-antigen polysaccharide of serotype-5 lipopolysaccharide. Analysis of these results revealted that the O-antigen polysaccharide of lipopolysaccharide was 1 of the antigenic determinants responsible for the serotype specificity of A pleuropneumoniae. On the other hand, 5MAb-6 reacted with strains of serotype 1 through 10 in varying degrees and its epitope was located on polypeptides sensitive to proteinase K. In an immunoblotting analysis, 5MAb-6 reacted with 2 polypeptide bands, with molecular weights of approximately 41,500 and 28,000, in the outer membrane protein-rich fraction obtained from strains of serotype 1 through 10. These results indicated that outer membrane proteins from several serotype strains of A pleuropneumoniae possessed common antigenic determinants.

SUMMARY

Two monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5, designated as 5MAb-l and 5MAb-6, were characterized. Enzyme-linked immunosorbent assay-inhibition tests with whole-cell antigens obtained from strains of serotype 1 through 12 of A pleuropneumoniae revealed that 5 MAb-1 bound to only serotype-5 strains. The epitope recognized by 5MAb-l was a carbohydrate that was sensitive to periodate oxidation and resided on the structure of β-l,6-linked d-galactose in an O-antigen polysaccharide of serotype-5 lipopolysaccharide. Analysis of these results revealted that the O-antigen polysaccharide of lipopolysaccharide was 1 of the antigenic determinants responsible for the serotype specificity of A pleuropneumoniae. On the other hand, 5MAb-6 reacted with strains of serotype 1 through 10 in varying degrees and its epitope was located on polypeptides sensitive to proteinase K. In an immunoblotting analysis, 5MAb-6 reacted with 2 polypeptide bands, with molecular weights of approximately 41,500 and 28,000, in the outer membrane protein-rich fraction obtained from strains of serotype 1 through 10. These results indicated that outer membrane proteins from several serotype strains of A pleuropneumoniae possessed common antigenic determinants.

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 5661 5632 1470
PDF Downloads 42 33 2
Advertisement