GENETIC-VARIATION OF HIV TYPE-1 IN 4 WORLD-HEALTH-ORGANIZATION-SPONSORED VACCINE EVALUATION SITES - GENERATION OF FUNCTIONAL ENVELOPE (GLYCOPROTEIN-160) CLONES REPRESENTATIVE OF SEQUENCE SUBTYPE-A, SUBTYPE-B, SUBTYPE-C, AND SUBTYPE-E

As part of the WHO Network for HIV Isolation and Characterization, we PCR amplified, cloned, and sequenced gp120 and gp160 genes from 12 HIV -1 isolates collected in four WHO-sponsored vaccine evaluation sites ( Brazil, Rwanda, Thailand, Uganda). Envelope clones were derived from P BMC-grown isolates obtained from asymptomatic individuals within 2 yea rs of seroconversion. Analysis of their deduced amino acid sequences i dentified all but one to contain an uninterrupted open reading frame. Transient expression and biological characterization of selected gp160 constructs identified six clones to encode full length and functional envelope glycoproteins. Phylogenetic analysis of their nucleotide seq uences revealed that they represent HIV-1 subtypes A, B, C, and E. Sin ce current knowledge of HIV-1 envelope immunobiology is almost exclusi vely derived from subtype B viruses, these reagents should facilitate future envelope structure, function and antigenicity studies on a broa der spectrum of viruses. This should assist in the design and evaluati on of effective vaccines against HIV-1.