AEGiS-12IAC: Tropism and co-receptor usage of tissue-derived HIV-1 primary isolates obtained from patients at the terminal stage of disease.

12th International AIDS Conference

Geneva, Switzerland - June 28-July 3, 1998

Tropism and co-receptor usage of tissue-derived HIV-1 primary isolates obtained from patients at the terminal stage of disease.

Int Conf AIDS 1998 Jun 28-Jul 3; 12:5 (abstract no. 11116)

Naif H, Li S, Alali M, Wasr N, Cunningham A
Centre for Virus Research Westmead Hospital, Sydney, NSW, Australia.

OBJECTIVES: To investigate the tropism and replication characteristics of HIV-1 isolates derived from different tissues (brain, CSF, lung, and spleen) in human monocytes and in vitro cultured monocyte-derived macrophages (MDM). Differential tropism and level of replication of these isolates was analysed in association with the co-receptor usage and the state of maturation/differentiation of monocytes into MDM.

METHODS: 10 tissue-derived HIV-1 isolates from brain, spleen, lung and cerebrospinal fluids and HIV-1 BaL were subcultured on PBMCs then used to infect fresh monocytes, monocyte-derived macrophages (MDM) and primary T lymphocytes. Replication kinetics of HIV-1 isolates was measured by p24 antigen ELISA and hot-PCR. V3 regions from viral DNA outputs were amplified using nested-PCR. Co-receptor usage was monitored by HOS cells transfected with cDNA for chemokine receptors.

RESULTS: Three levels of replication were observed: high, low or non-productive in both monocytes and MDM while only two levels (high and low) were observed in T lymphocytes. Most isolates (7/10) replicated with productive infection in both monocytes and MDM as tested by the extracellular p24 antigen. Full length HIV DNA was correlated with the level of extracellular p24 antigen in most isolates, as detected by highly sensitive hot-PCR. The level of restriction of low replicating isolates was prior to reverse transcription. V3 sequences revealed that these isolates had similar genotypes to those of non-syncytium inducing (NSI) macrophage-tropic strains and phylogenetic analysis demonstrated a clear segregation of isolates grown on different cell types especially MDM. Most isolates used the chemokine receptor 5 (CCR5) to infect cells as determined by infection assay using HOS cells.

CONCLUSIONS: Tissue-derived HIV isolates replicated either at low or high level of replication irrespective of the state of cell maturation. High replicating isolates showed no restriction at any stage of replication and low replicating isolates restricted at the stage of pre-reverse transcription. Most isolates used CCR5 as the main co-receptor to enter cells although they have been obtained at the terminal stage of disease.

Keywords: AEGIS, HIV-1, Tropism, Virus Replication, Macrophages, Monocytes, HIV Infections, Receptors, Chemokine, Giant Cells, DNA, Viral, Genotype, Polymerase Chain Reaction, Greece, Human, In Vitro, virology, genetics, ICA12
980628
11116