Inhibition of M-tropic HIV-1 infection by the fd phage-gene 3 protein with MIP-1alpha-binding activity.
CCR5 is a chemokine receptor with seven transmembrane-domains. It is expressed on T cells and macrophages and functions as the principal co-receptor for macrophage (M)-tropic strains of HIV-1. The anti-CCR5 monoclonal antibody (mAb) 2D7 inhibits the binding and chemotaxis of the three natural beta-chemokine ligands of CCR5, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES, to CCR5(+) cells. The mAb also efficiently blocks the infectivity of several M-tropic and dual-tropic HIV-1 strains in vitro. In this study, we attempted to determine the peptide motif recognized with the 2D7 mAb. We isolated phage clones by panning a phage display library using 2D7 and identified three peptide motifs. One of these phage clones (M23) showed a marked inhibitory activity on HIV-1 infection. The unique sequence of 15 amino acids with an internal disulfide bond was inserted in the g3p of the M23 phage clone (M23-g3p). The M23-g3p was purified by fast-performance liquid chromatography (FPLC). We show here that (1) M23-g3p was specifically recognized with anti-CCR5 mAb; (2) M23-g3p showed inhibitory activity on the infectivity of M-tropic but not T-tropic HIV-1 strains; (3) M23-g3p bound to MIP-1alpha, MIP-1beta, and RANTES but not MCP-1. These results suggested that the M23-g3p might mimic the CCR5-binding domain shared by beta-chemokines, MIP-1alpha, MIP-1beta, and RANTES as well as the HIV-1 infection.
Meta
A
A
Department of Bioengineering, Faculty of Engineering, Kagoshima University, 1-21-40 Korimoto, Kagoshima, Japan.
Torigoe
N
N
Ito
Y
Y
Arakaki
R
R
Nakashima
H
H
Sugimura
K
K
D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity; D001483:Base Sequence; D036022:Capsid Proteins; D002460:Cell Line; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000494:pharmacology*; D015658:HIV Infections / Q000517:prevention & control*; D015497:HIV-1 / Q000472:pathogenicity; D006801:Humans; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology / Q000821:virology; D051379:Mice; D008969:Molecular Sequence Data; D019713:Receptors, CCR5 / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000821:virology; D014760:Viral Fusion Proteins / Q000378:metabolism* / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D017931:DNA Primers; D004268:DNA-Binding Proteins; D019402:Macrophage Inflammatory Proteins; D019713:Receptors, CCR5; D014760:Viral Fusion Proteins