Both in vitro culture and in vivo passage lead to selective replication of SIV variants.

Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.

Both in vitro culture and in vivo passage lead to selective replication of SIV variants.

Symp Nonhum Primate Models AIDS. 1990 Nov 28-30;8:18 (abstract no. 2). Unique Identifier : AIDSLINE PRIM8/900002
Villinger F; De B; Jehuda-Cohen T; Powell JD; Neckelmann N; McClure HM; Folks TM; Ansari AA; Yerkes Regional Primate Research Center, Dept. of Pathology,; Emory University School of Medicine, Atlanta, GA 30322

Abstract: A set of oligo primers was prepared based on the sequence shared by various SIV isolates from a variety of nonhuman primate species. This primer pair was used to amplify and sequence the middle third of the gag region of the endogenous SIVsmm (SIVsmm-e) present in the DNA prepared from the PBMC of naturally infected clinically asymptomatic sooty mangabeys. Comparison of the aligned sequences of 8 PCR-derived clones from 2 different mangabeys revealed a modest 3% variation in the nucleotide sequence. A similar comparison of this endogenous mangabey SIV nucleotide sequence with the published sequence of SIVsmmh4 revealed 88% homology. Of interest was the observation of a high degree of divergence within a short gag sequence (nt 1856-1880) between the endogenous mangabey SIV (SIVsmm-e) and the published sequence of SIVsmmh4, which corresponded to a similar degree of divergence between SIVsmmh4 and the published sequence of SIVmac251 (SIVmm251). An oligo nucleotide probe (probe 1) based on the SIVsmmh4 sequence and encompassing this highly divergent gag region was found to hybridize with PCR products amplified from the DNA prepared from PBMC of 14 of 16 macaques experimentally infected with SIVsmm-9 (a tissue culture isolate from a sooty mangabey) and with tissue culture cell lines infected in vitro with SIVsmm-9. However, PCR products amplified from the DNA prepared from PBMC of 41 infected sooty mangabeys failed to hybridize with this SIVsmmh4-specific probe (probe 1) but did hybridize with a probe based on the corresponding sequence present in SIVsmm-e (probe 2). Of further interest was the observation that DNA from the PBMC (of 3 sooty mangabeys) which had been cultured in vitro for 7 days yielded PCR products which did hybridize with probe 1. Additional significance to these findings is provided by results of PCR studies on DNA samples prepared from PBMC of infant macaques which were offspring of macaques infected with SIVsmm-9 at varying times during pregnancy. PCR products amplified from the DNA of PBMC of 10 of the 11 offspring macaques hybridized with oligo probe 2 (SIVsmm-e specific) but not with the oligonucleotide probe 1 (SIVsmmh4 specific) while PCR products amplified from the DNA of PBMC of all the experimentally infected mothers hybridized with probe 1 and some with both probes 1 and 2. These data cumulatively suggest that the endogenous SIV present in asymptomatic sooty mangabeys may be distinct from the in vitro tissue culture propagated SIVsmm-9 and that isolated at the Delta Primate Center and sequenced by Drs. Hirsch and Johnson (in vitro selection). In addition, these data suggest that the perinatal mode of transmission results in the selective replication of a variant of SIV (in vivo selection) in the offspring.

Keywords: Animal *DNA Replication DNA, Viral/BIOSYNTHESIS/*CHEMISTRY Female Genes, gag In Vitro Leukocytes, Mononuclear/CHEMISTRY Macaca Oligonucleotide Probes Polymerase Chain Reaction Pregnancy Sequence Homology, Nucleic Acid SIV/GROWTH & DEVELOPMENT/*GENETICS Transfection Variation (Genetics) *Virus Replication ABSTRACT
910730
M9170992

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