Monoclonal antibodies to Pasteurella haemolytica serotype 1 lipopolysaccharide: demonstration of antigenic similarities among several serotypes

Abstract

Murine monoclonal antibodies (MAbs) were produced which were specific for Pasteurella haemolytica serotype 1 lipopolysaccharide (LPS). The MAbs also reacted with LPS present in a partially purified antigen derived from a saline extract of the organism. The epitope to which the MAbs were directed was a carbohydrate which was sensitive to oxidation with periodate, had a molecular weight between 14,000 and 25,000 as determined by immunoblotting, and was present in a crude 0-antigen preparation of P. haemolytica LPS. The MAbs did not react with purified capsular polysaccharide from P. haemolytica serotype 1. In an enzyme-linked immunosorbent assay, reaction of the MAbs with LPS obtained from 14 gram-negative bacteria failed to detect any cross-reactivity with P. haemolytica LPS. However, the MAbs detected antigenic similarities among P. haemolytica serotypes 1, 5, 6, 7, 8, and 12 and, to a lesser extent, 4 and 14. These studies indicate that the LPS-O-antigens from several P. haemolytica serotypes have similar epitopes and may be partially responsible for shared antigenicity among serotypes.

Structural and serological specificities of Pasteurella haemolytica lipopolysaccharides

“…LPS has been implicated as a major antigenic determinant of P. haemolytica (10), and antigenic similarities among the LPSs of several P. haemolytica serotypes have been reported (6,13,16,36,45). P. haemolytica serotypes belonging to biotype T have been observed to elaborate LPS having extensive 0-chains (28,33,36).…”

“…(35). Serotypes 1,5,6,7,8,12,14,and 16 share common core oligosaccharide antigenic determinants as determined by SDS-PAGE immunoblots with polyclonal sera or monoclonal antibody with specificity to epitope(s) in the type 1 core region. Immunoblots with serotype 2 antiserum revealed a low level of reaction with all other serotypes in the core region, which possibly indicates that this strain contains a common but weakly immunogenic antigen.…”

“…Silver-stained SDS-PAGE profiles of P. haemolytica LPS demonstrating core oligosaccharide and O-polysaccharide regions for16 serotypes. Lanes 1 to 16 are PK digests of serotypes 1 to 16, respectively.…”

“…to 16, respectively. Comparison of antigenic specificities of core and O-polysaccharide LPS components of the 16 serotypes of P. haemolytica identified from immunoblots Represents serotypes exhibiting shared core antigens as determined from antisera in stub.…”

Production and specificity of poly‐ and monoclonal antibodies raised against Shewanella putrefaciens

“…There are a number of reports that describe the raising, testing and usage of MAbs against bacteria or their products. Purified antigens have been used for the immunization in the majority of studies (Galloway et al 1984;Durham et al 1988;Yamaguchi et al 1989;Kabir 1991;Choi et al 1992;Daya-Mishne and Shabtai 1992). Antibodies against whole cells have been produced successfully (Sugasawara et al 1983;Bogard et al 1987;Applemelk et al 1988;Vernon-Shirley and Burns 1992).…”