AEGiS-12IAC: Genetic and biological characterization of Russian HIV-1 subtype B isolate with unusual V3 loop.

12th International AIDS Conference

Geneva, Switzerland - June 28-July 3, 1998

Genetic and biological characterization of Russian HIV-1 subtype B isolate with unusual V3 loop.

Int Conf AIDS 1998 Jun 28-Jul 3; 12:23 (abstract no. 11204)

Khanina T, Selimova LM, Serebrovskaya LV, Kravchenko AV, Pokrovsky VV
D.I. Ivanovsky Institute of Virology, Moscow, Russia.

OBJECTIVES: To investigate Russian HIV-1 subtype B isolate with unusual V3 loop.

METHODS: The patient RU0025 was a homosexual male living in Moscow. He was tested HIV-1-positive in December 1991. Owing to disease progression patient accepted AZT therapy continually from May 1992. In June 1995, periferal blood mononuclear cells (PBMCs) were separated using Ficoll-gradient centrifugarion. At that time the CD4 count was 0.14. To isolate virus, patient's PBMCs were co-cultivated with donors' PBMCs preliminary stimulated for 3 days with phytohemagglutinin (3 ug/mL). PBMCs cultures were maintained in RPMI 1640 medium containing 10% fetal calf serum, interleukin-2 (10 IU/mL), and antibiotics. After 4 subsequent co-cultivations the biological phenotype of the isolate, named 95RU25B, was determined, and the 0.4 kbp env gene fragment encompassing the C2-V3 region of gp120 was amplified using polymerase chain reaction, cloned in pUC18 vector plasmid, and sequenced.

RESULTS: The isolate 95RU25B formed syntitia in SupT1 cells and was able to grow in U937.2 cell line, displaying S/L.3 phenotype. Genetic analysis of 95RU25B revealed that this isolate belonged to HIV-1 env genetic subtype B. However, genetic distance between 95RU25B and the subtype B prototypic strains, MN and JRCSF, was 11.3% and 12.9%, respectively. Moreover, the translated amino acids sequence of the V3 loop had three residues insertion as well as three residue deletion situated upstream and downstream of the crown of V3, respectively: ctrpnnntrkririGHIgpgrafyat ... igdirqahc.

CONCLUSION: The ability of 95RU25B to grow both in SupT1 and U937.2 cell lines may result from unusual structure of the V3 loop.

Keywords: AEGIS, HIV-1, Genes, env, HIV Infections, HIV Seropositivity, Amino Acid Sequence, CD4 Lymphocyte Count, Phenotype, Polymerase Chain Reaction, Moscow, Greece, Male, Human, genetics, ICA12
980628
11204