Characterization of the African HIV-1 isolate CBL-4 (RUT) by partial sequence analysis and virus neutralization with peptide antibody and antisense phosphate-methylated DNA.
The HIV-1 isolate CBL-4 (RUT), originating from Tanzania, was characterized using a comprehensive virus-typing system. This system included sequence analysis of the region coding for the neutralization domain in the third variable region (V3) of the external envelope and of the tat responsive (TAR) region after polymerase chain reaction (PCR) amplification of these sequences from cellular DNA in the CBL-4 (RUT) producer line. Based on independent cluster analysis of TAR and V3 sequences the CBL-4 (RUT) virus was positioned closest to the Z6 (and ELI) African virus family. The V3 amino acid sequence on the surface of the virus particle was confirmed by the inhibition of neutralization of CBL-4 (RUT) by a synthetic peptide derived from the nucleic acid sequence. Using antisense phosphate-methylated DNA covering the TAR loop region of LAV-1/HTLV-IIIB, inhibition of HTLV-IIIB and HTLV-IIIRF infection was seen, whereas no inhibition was observed for CBL-4 (RUT), indicating two or more mismatches in the TAR loop region, a characteristic shared with Z6 virus, but not with ELI. We propose a virus-typing system based on sequence analysis confirmed by virus neutralization with a peptide binding antibody and inhibition by antisense phosphate-methylated DNA to group viruses for laboratory use and vaccine design.
Human Retrovirus Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands.
J
J
Goudsmit
J
J
Geelen
W
W
Keulen
D
D
Notermans
C
C
Kuiken
C
C
Ramautarsing
L
L
Smit
L
L
Koole
M
M
van Genderen
H
H
Buck
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D004279:DNA, Viral / Q000378:metabolism; D015497:HIV-1 / Q000145:classification* / Q000235:genetics; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008745:Methylation; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D012703:Serotyping; D014765:Viral Vaccines / Q000276:immunology
D000906:Antibodies; D004279:DNA, Viral; D007135:Immunoglobulin Variable Region; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D014765:Viral Vaccines