A cyclic dodecapeptide-multiple-antigen peptide conjugate from the undecapeptidyl arch (from Arg(168) to Cys(178)) of extracellular loop 2 in CCR5 as a novel human immunodeficiency virus type 1 vaccine.
A cyclic closed-chain dodecapeptide (cDDR5) mimicking the conformation-specific domain of CCR5 was prepared in which Gly-Asp, as a dipeptide forming a spacer arm, links the amino and carboxyl termini of the decapeptidyl linear chain (Arg(168) to Thr(177)) derived from the undecapeptidyl arch (UPA; Arg(168) to Cys(178)) of extracellular loop 2 (ECL2) in CCR5. Novel monoclonal antibodies were raised against cDDR5 conjugated with a multiple-antigen peptide (cDDR5-MAP), and the purified antibody [KB8C12, immunoglobulin M(kappa)] reacted with cDDR5, but not with linear DDR5, in real-time biomolecular interaction analysis using surface plasmon resonance. The antibody also reacted with cells expressing CCR5, but not with cells expressing CXCR4, and the immunoreaction was competed by cDDR5-MAP. The antibody significantly interfered with chemotaxis induced by macrophage inflammatory protein, 1beta, and at a concentration of 1.67 nM it almost completely inhibited infection by human immunodeficiency virus type 1 (HIV-1) R5, but not by HIV-1 X4, as observed by use of a new phenotypic assay for drug susceptibility of HIV-1 using the CCR5-expressing HeLa CD4(+) cell clone 1-10 (MAGIC-5). Furthermore, cDDR5-MAP suppressed infection by HIV-1 R5 at relatively high concentrations (50 to 400 microM) in a dose-dependent manner but did not suppress infection by HIV-1 X4. Taken together, these results indicate that the antibody is conformation specific and recognizes the conformation-specific domain of the UPA of ECL2. Moreover, both the antibody and its immunogen, the cDDR5-MAP conjugate, may be useful in developing a new candidate vaccine for HIV therapy.
Misumi
S
S
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan.
Nakajima
R
R
Takamune
N
N
Shoji
S
S
D016915:AIDS Vaccines / Q000737:chemistry / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001120:Arginine / Q000737:chemistry*; D054407:Chemokine CCL4; D002633:Chemotaxis / Q000187:drug effects; D003545:Cysteine / Q000737:chemistry*; D005434:Flow Cytometry; D015488:HIV Antigens / Q000737:chemistry / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D019402:Macrophage Inflammatory Proteins / Q000494:pharmacology; D008561:Membrane Fusion / Q000276:immunology; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology*; D019713:Receptors, CCR5 / Q000737:chemistry / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D054407:Chemokine CCL4; D015488:HIV Antigens; D019402:Macrophage Inflammatory Proteins; D009842:Oligopeptides; D019713:Receptors, CCR5; D001120:Arginine; D003545:Cysteine