Construction of recombinant targeting immunogens incorporating an HIV-1 neutralizing epitope into sites of differing conformational constraint.
2F5 is one of the very few monoclonal antibodies with the capacity to neutralize a wide spectrum of type 1 human immunodeficiency virus (HIV-1) strains and primary isolates. Constructing an immunogen that contains a conformational mimic of the epitope recognized by 2F5 could provide the means to induce a broadly neutralizing anti-HIV-1 antibody response. Thus, in an effort to create a targeted, adjuvant-independent immunogen able to induce a 2F5-like antibody response, the gp41 sequence recognized by 2F5 (ELDKWAS) was genetically incorporated into different regions of an antibody specific for a framework determinant on human leukocyte antigen (HLA)-DR. All constructs were expressed, secreted from Sf9 insect cells, and found to retain the anti-HLA-DR specificity of the parental antibody. Three of the four constructs in which the ELDKWAS sequence was incorporated into a beta-turn (BT)-like conformational site were recognized by the 2F5 antibody. In contrast, none of the five constructs with the same sequence incorporated into surface-exposed regions of helical turn had any detectable 2F5 reactivity. In addition to demonstrating the significant plasticity of several regions in the antibody molecule in terms of accepting foreign sequences without loss of expression or binding specificity, these results also suggest that the native epitope recognized by the 2F5 antibody may be more beta-turn-like than helical in conformation. Importantly, with respect to vaccine development, the 2F5-reactive antibody constructs represent candidate immunogens for the adjuvant-independent induction of an HIV-1, neutralizing 2F5-like antibody response in humans.
Ho
Jason
J
Department of Immunology, Medical Sciences Building, 1 Kings College Circle, University of Toronto, Ont., M5S 1A8, Toronto, Canada.
MacDonald
Kelly S
KS
Barber
Brian H
BH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D000939:Epitopes; D015483:HIV Antibodies / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006684:HLA-DR Antigens / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D011994:Recombinant Proteins