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A hamster anti-idiotype monoclonal antibody, mimicking the inner-core region of Gram-negative bacterial lipopolysaccharide (LPS), stimulates LPS inner-core-specific serum antibodies in hamsters

Department of Microbiology, Molecular Genetics and Immunology, and the Cancer Center, University of Kansas Medical Center, Kansas City, USA

D.C. Morrison

Department of Microbiology, Molecular Genetics and Immunology, and the Cancer Center, University of Kansas Medical Center, Kansas City, USA

We have earlier described the development of an Ab2β anti-idiotype antibody specific for idiotypic determinants of a BALB/c mouse IgG3 monoclonal antibody (MAbY1-4A6) directed against the conserved inner-core 2-keto-3-deoxyoctulosonate (Kdo) residue of Salmonella minnesota rough mutant Re lipopolysaccharide (Re-LPS) [Field S, Pollack M, Morrison D C. Microb Pathog 1993; 15: 103-120]. Immunization of both inbred and outbred mice with MAb4G2 protected these animals against subsequent lethal challenge with S. minnesota Re-LPS. Here we demonstrate that hamsters immunized with the anti-idiotype MAb4G2 generate serum antibodies which cross-react with a variety of forms of LPS, as evidenced by specific dose-dependent binding to R and S chemotype LPS as assessed by ELISA and specific binding to antibiotic-treated, whole bacterial culture supernatants and to LPS-containing fractions in sucrose gradient velocity-sedimentation-fractionated, antibiotic-treated bacterial culture supernatants. Competitive inhibition assays using MAb4G2-elicited hamster antibodies (Ab3 or Ab1') and the Kdo-specific MAbY1-4A6 (Ab1) were carried out to assess the Kdo epitope-specificity and Ab1'-like properties of the hamster MAb4G2 antibodies (Ab3). Hamster MAb4G2 serum antibodies and the Ab1 MAbY1-4A6 express similar epitope specificities for both the MAb4G2 and Re-LPS antigens, confirming the Kdo epitope-specificity of these Ab1' serum antibodies. These data further establish that MAb4G2 is a true internal surrogate image of the conserved inner-core Kdo epitope of LPS.

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