Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.
Characterization of SIV envelope proteins derived from SIVmac infected macaques shortly after infection.
Symp Nonhum Primate Models AIDS. 1992 Nov 17-20;10:abstract no. 117. Unique Identifier : AIDSLINE PRIM10/93200950 Hulskotte EG; Rimmelzwaan GF; Bosch ML; de Vries P; Heeney JL; Osterhaus AD; Laboratory of Immunobiology, RIVM, Bilthoven, The Netherlands.
Abstract:
We have shown that vaccination with an ISCOM preparation based on envelope protein enriched whole SIVmac is effective against subsequent challenge with the homologous strain of SIV. It is of interest to know whether envelope protein alone incorporated into ISCOMs would also be able to confer protection against SIV challenge. Variation in the HIV/SIV envelope proteins has been shown to be important in pathogenesis and disease progression and may also be expected to interfere with anti-viral immunity. Therefore it is of interest to gain insight in the level of heterogeneity of the genotypes that establish SIV infection. A nested-primer polymerase chain reaction (PCR) was used to amplify the envelope gene directly from PBMCs of a rhesus macaque (#8789) two weeks after infection with the 32H isolate of SIVmac251. This isolate has been shown to contain a heterogeneous population of viruses, which exhibited the highest degree of variation in the variable regions 1 and 2 (V1 and V2) of the envelope sequence. PCR products were cloned and the nucleotide sequence of five out of six envelope clones proved to be completely identical in the V1 and V2 regions. Data obtained from the analysis of envelope clones recovered from another rhesus macaque (#8672), which was infected with PBMCs isolated from a SIVmac(32H) infected rhesus monkey 11 months after infection, exhibited the same degree of homology among SIV envelope sequences recovered shortly after infection. These data suggest that infection with SIV is established by a limited number of the genotypes present in the inoculum. One envelope clone from each monkey, designated E8789 and E8672 respectively, was selected and their complete nucleotide sequences were determined. Comparison of the two sequences showed amino acid differences throughout the envelope proteins, however, a clustering of amino acid differences was observed in the V1 region. The two envelope genes cloned in E8789 and E8672 were expressed in a recombinant vaccinia virus expression system. For cloning into vaccinia virus a plasmid (pSC65) was used containing vaccinia TK sequences flanking the cloning site, the lacZ gene and a synthetic early/late vaccinia promotor under the control of which the SIV envelope genes were cloned. In addition recombinant vaccinia viruses were constructed that express the SIV envelope gene of which the region coding for the cleavage site between gp120 and gp41 had been mutated by site directed mutagenesis. The expressed SIV glycoproteins were biochemically characterized and are currently used in the formulation of a candidate SIV vaccine.
Keywords: beta-Galactosidase/GENETICS Animal Base Sequence Cloning, Molecular Comparative Study Gene Products, env/*GENETICS/ISOLATION & PURIF Genes, env Genotype Lymphocytes/*MICROBIOLOGY Macaca mulatta Mutagenesis, Site-Directed Polymerase Chain Reaction/METHODS Promoter Regions (Genetics) Sequence Homology, Amino Acid Simian Acquired Immunodeficiency Syndrome/IMMUNOLOGY/ *MICROBIOLOGY SIV/GENETICS/*ISOLATION & PURIF Thymidine Kinase/GENETICS Vaccinia Virus/GENETICS ABSTRACT 930630
M9361070
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Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.
