Dependence of the murine antibody response to an anti-CD4 CDR2 V H peptide on immunogen formulation

A peptide corresponding to the second complementarity determining region of the heavy chain (CDR2 V H) from a murine anti-CD4 monoclonal antibody, designated L202, was synthesized by solid phase methodology in a number of different antigenic forms, for the purpose of comparing the effectiveness of d...

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Bibliographic Details
Published inMolecular immunology Vol. 32; no. 17; pp. 1319 - 1328
Main Authors Kanda, Patrick, Fritz, David A., Gage, Douglas A., Shuler, Kevon R.
Format Journal Article
LanguageEnglish
Published Elsevier Ltd 01.12.1995
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Summary:A peptide corresponding to the second complementarity determining region of the heavy chain (CDR2 V H) from a murine anti-CD4 monoclonal antibody, designated L202, was synthesized by solid phase methodology in a number of different antigenic forms, for the purpose of comparing the effectiveness of different adjuvant-carrier systems in the induction of a murine antibody response against the immunizing peptide and parent antibody molecule. Two of the synthetic constructs contained the palmitoyl and N-palmitoyl-cysteinyl- S-(2,3-palmitoyloxy)-propanediol (PAM 3Cys) moieties, respectively, attached to the peptide amino terminus with the immunogen comprising liposomal formulations of each. A third immunogen consisted of the CDR2 V H peptide admixed with the PAM 3Cys non-covalently and incorporated into liposomes (PAM 3Cys + CDR2 V H). A fourth composition comprised the CDR2 V H peptide conjugated to KLH via the sulfhydryl of an added N terminal cysteine (KLH-CDR2 V H) and injected with Complete Freund's adjuvant (CFA). A fifth immunogen consisted of the CDR2V H peptide synthesized on an octameric, branched polylysine core as a multiple antigenic peptide (MAP-CDR2 V H) injected in the presence of Freund's adjuvant. Groups of five mice were injected intramuscularly with each of these immunogens and bled at two week intervals. The highest anti-peptide gamma-immunoglobulin (IgG) responses (against uncoupled peptide by ELISA) after 56 days were obtained with mice receiving the PAM 3Cys-CDR2 V H peptide. However, when screened against the CDR2 V H peptide present as the MAP derivative by ELISA, IgG raised against the cognate MAP-CDR2 peptide was much more reactive than IgG raised against the liposomal PAM 3Cys-CDR2 V H immunogen. In either case, IgG raised against the KLH-CDR2V H conjugate was poorly reactive. These differences in reactivity to the two forms of the CDR2 V H peptide by ELISA did not correspond to major differences in reactivities to the intact L202 Ab by ELISA. Although the IgG against the MAP immunogen was slightly more reactive than the other antisera against the L202 Ab, all titers were less than 1:100. These data illustrate some limitations of using anti-peptide responses as indicators of potential reactivity against the native protein, but suggest that alternate formulations including lipoidal peptides are more effective than corresponding KLH-peptide conjugates in eliciting Ab responses against poorly immunogenic epitopes.
ISSN:0161-5890
1872-9142
DOI:10.1016/0161-5890(95)00117-4