The construction and characterization of poliovirus antigen chimeras presenting defined regions of the human T lymphocyte marker CD4.
Defined regions of the CDR2-like region of the T cell antigen CD4 that are implicated in the binding of the surface glycoprotein (gp120) of human immunodeficiency virus type 1 (HIV-1) to CD4+ T lymphocytes have been engineered in place of antigenic site 1 of Sabin type 1 poliovirus. The antigenic properties of the recovered chimeric virus particles were investigated using monoclonal antibodies (MAbs) and polyclonal serum to CD4. None of the MAbs tested neutralized the chimeras, presumably because they are directed against conformational determinants of the V1 domain of CD4. In contrast, the three antigen chimeras were neutralized by polyclonal serum to CD4, which suggested that the CD4-derived sequences were presented in a relevant conformation. A panel of six MAbs were raised against one of the chimeras, and the epitopes were mapped by the selection of neutralization-resistant mutants and cross-neutralization studies. Five of the six MAbs reacted with soluble CD4 (sCD4) in ELISA, and one (MAb 1686) bound to CD4 expressed at the surface of HeLa cells. The high affinity interaction between gp120 and sCD4 was not blocked by MAb 1686, and the poliovirus-CD4 chimeras did not interact with gp120. These results demonstrate that poliovirus can be used as an epitope expression vector for the presentation of sequences in an immunodominant location on the virus particle which adopt a native or near-native conformation, and supports the findings of previous studies involving the presentation of epitopes derived from pathogens.
Rose
C
C
Department of Microbiology, University of Reading, Whiteknights, U.K.
Andrews
W
W
Ferguson
M
M
McKeating
J
J
Almond
J
J
Evans
D
D
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D000918:Antibody Specificity; D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D017955:Poliovirus / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D036022:Capsid Proteins; D015699:HIV Envelope Protein gp120; D011993:Recombinant Fusion Proteins; C442540:VP1 protein, Poliovirus