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Monoclonal ligand binding site related anti-idiotypic antibodies elicited with a polyclonal kinin antibody
C E Odya1, R Yapa, B Soltani-Tehrani
1Medical Sciences Program, Indiana University School of Medicine, Bloomington 47405.
Splenocytes from mice immunized with homogenous, polyclonal, rabbit kinin antibody (BK21) were fused using polyethylene glycol with the mouse myeloma cell line SP2/o. Eleven monoclonal antibodies, whose binding to BK21 could be inhibited by bradykinin, were obtained from 3 fusions. All of these anti-idiotypic antibodies were of the IgG1k isotype, except for one, which was an IgG2ak. An IgMk, auto-anti-idiotypic antibody, reactive with BK21 was obtained from a fusion of SP2/o cells and splenocytes from a mouse immunized with bradykinin conjugated with carbodiimide to keyhole limpet hemocyanin. Bradykinin could completely inhibit the binding of all of the anti-idiotypic antibodies to BK21 in an enzyme-linked immunosorbent assay. This result is consistent with the anti-idiotypic antibodies being reactive with the ligand binding sites of BK21. It was possible to separate the anti-idiotypic antibodies into 2 groups. The first group, 10 of the 12 antibodies tested, was more sensitive to inhibition by bradykinin than the second group and was not readily inhibited by des-Arg9-bradykinin. The second group was about 7 times more sensitive to inhibition by des-Arg9-bradykinin than by bradykinin. Further experiments will be needed to determine whether or not these monoclonal anti-idiotypic antibodies are "internal image" antibodies.
Splenocytes from mice immunized with homogenous, polyclonal, rabbit kinin antibody (BK21) were fused using polyethylene glycol with the mouse myeloma cell line SP2/o. Eleven monoclonal antibodies, whose binding to BK21 could be inhibited by bradykinin, were obtained from 3 fusions. All of these anti-idiotypic antibodies were of the IgG1k isotype, except for one, which was an IgG2ak. An IgMk, auto-anti-idiotypic antibody, reactive with BK21 was obtained from a fusion of SP2/o cells and splenocytes from a mouse immunized with bradykinin conjugated with carbodiimide to keyhole limpet hemocyanin. Bradykinin could completely inhibit the binding of all of the anti-idiotypic antibodies to BK21 in an enzyme-linked immunosorbent assay. This result is consistent with the anti-idiotypic antibodies being reactive with the ligand binding sites of BK21. It was possible to separate the anti-idiotypic antibodies into 2 groups. The first group, 10 of the 12 antibodies tested, was more sensitive to inhibition by bradykinin than the second group and was not readily inhibited by des-Arg9-bradykinin. The second group was about 7 times more sensitive to inhibition by des-Arg9-bradykinin than by bradykinin. Further experiments will be needed to determine whether or not these monoclonal anti-idiotypic antibodies are "internal image" antibodies.