Immune recognition of porin and lipopolysaccharide epitopes of Salmonella typhimurium in mice.
We investigated the antigenic specificity of the humoral immune response to infection by Salmonella typhimurium, by competitive inhibition enzyme-linked immunosorbent assay and Western immunoblots. A panel of eight murine monoclonal antibodies, raised to OmpC and OmpD porins and lipopolysaccharide (LPS)-O antigens, was used to define the specificity of the polyclonal immune response in mice. The monoclonal antibody panel recognized five distinct epitopes; these were localized to surface-exposed loops of OmpC and OmpD porin, to the "eye-let" forming loop L3 of OmpC/OmpD, and to LPS-O4 and O5 factors. The immune mouse serum raised to infections with S. typhimurium LT-2 strain WB600 (wild-type) competitively inhibited the binding of biotin-labelled monoclonal antibodies to the epitopes that they recognize, indicating that all five epitopes were targets of the host immune response to natural infection. However, only two epitopes, one within a surface-exposed loop of OmpC porin, and the other in the LPS-O4 factor, were immunodominant. Furthermore, the bacterial LPS core and O-antigen structure influenced the immune response to the porins. Surface epitopes of porins were dominant in the rough strain SH5014 (rfa), whereas the immune recognition of LPS epitopes was predominant in mice infected with the smooth, wild-type strain (WB600). Finally, the immune response to LPS epitopes O4 and O5 was more pronounced in mice immunized with heat-killed cells than those infected with live S. typhimurium.
Singh
S P
SP
Biomedical Research and Training Programs, Alabama State University, Montgomery, AL 36101, USA. spsingh@asunet.alasu.edu
Williams
Y U
YU
Klebba
P E
PE
Macchia
P
P
Miller
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology; D016056:Immunodominant Epitopes / Q000032:analysis*; D008070:Lipopolysaccharides / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D018272:Porins / Q000276:immunology* / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000737:chemistry / Q000472:pathogenicity*
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D016056:Immunodominant Epitopes; D008070:Lipopolysaccharides; C113724:OmpC protein; D018272:Porins