Isolation and characterization of monoclonal antibodies to recombinant bovine interleukin-1 beta.
We describe the preparation of monoclonal antibodies (Mabs) directed against recombinant bovine interleukin-1 beta (rBoIL-1 beta). These anti-IL-1 beta Mabs were designated SA10, SA12, SA13, SA15, and SA22, and were characterized on the basis of their epitope specificity and cross-reactivity with homologous and heterologous cytokines in enzyme-linked immunosorbent assays and immunoblot analyses. Additionally, the ability of these Mabs to neutralize IL-1 beta was tested in thymocyte costimulation assays. The ELISA titers of all Mabs ranged from 9.4 x 10(6) to 1 x 10(7). Data indicate that Mabs SA10, SA12, SA15, and SA22 neutralized both bovine macrophage-derived IL-1 (1:4) and rBoIL-1 beta (1 ng ml-1). All the Mabs against rBoIL-1 beta (SA10, SA12, SA13, SA15, SA22) were specific and did not cross-react with other cytokines tested, except recombinant human IL-1 beta (rHuIL-1 beta). This finding suggests that these Mabs recognize epitopes common to human and bovine IL-1 molecules. Competition experiments suggested that Mab SA22 recognized a different epitope and Mabs SA10, SA12, SA13, and SA15 recognized the same epitope on the rBoIL-1 beta molecule. These observations suggest that these Mabs could be useful reagents for developing immunoassays to measure bovine IL-1 beta from biological fluids and to study the immunoregulatory role of IL-1 in the bovine immune system.
Reddy
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Department of Anatomy and Physiology, Kansas State University, Manhattan 66506.
Chitko-McKown
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Reddy
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Minocha
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HC
Blecha
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D001667:Binding, Competitive; D002417:Cattle / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D000939:Epitopes; D006825:Hybridomas; D015151:Immunoblotting / Q000662:veterinary; D007375:Interleukin-1 / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008809:Mice, Inbred C3H / Q000276:immunology; D009500:Neutralization Tests / Q000662:veterinary; D011863:Radioimmunoassay / Q000662:veterinary; D011994:Recombinant Proteins / Q000276:immunology; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007375:Interleukin-1; D011994:Recombinant Proteins