AEGiS-14IAC: cDNA expression array analysis of cell cycle regulation genes in CD4 and CD8 T lymphocytes from HIV-1 infected patients.

14th International AIDS Conference

Barcelona, Spain - July 7-12, 2002

cDNA expression array analysis of cell cycle regulation genes in CD4 and CD8 T lymphocytes from HIV-1 infected patients.

Int Conf AIDS 2002 Jul 7-12; 14:(abstract no. WeOrA1344)

Nobile M, Aschwanden E, Spasojevic M, Rizzardi GP, Khonkarly M, Pantaleo G
Laboratory of AIDS Immunopathogenesis, Division of Immunology and Allergy, Lausanne, Switzerland

BACKGROUND: Several mechanisms have been proposed to explain the functional and the quantitative defects of CD4 T cells with HIV-1 infection. At the present, limited information is available on the pattern of expression of genes involved in cell activation and in cell cycle. The cDNA expression array represents a comprehensive tool as it allows the simultaneous screening of large number of genes.

METHODS: Premade cDNA expression arrays spotted with 111 genes all related to cell cycle was used to screen CD4 and CD8 T cells in donors (n=8), in HIV-1 infected patients (CD4, n=8; CD8, n=10), and in vitro stimulated cells with anti-CD3 and anti-CD28 (n=3) during 60 hours. Expression was detected by radio labelling of cDNA by reverse transcription and exposure 6 hours of arrays to InstantImager. Data analysis was done with GeneCluster and TreeView softwares (Stanford Uni.).

RESULTS: An increased expression of the kinases, cyclins and CDKs gene families was observed in both CD4 and CD8 cells from healthy donors after in vitro stimulation consistently with cell activation and cycling activity. A similar pattern of gene expression was observed in CD8 T lymphocytes of HIV-1 infected subjects thus demonstrating increased activation and cycling activity in CD8 T cells. CD4 T cells had increased expression of cell activation genes (e.e. kinase genes) but lack expression of CDK2, a gene crucial for advancing cell cycle from the G1 to the S phase. Both CD4 and CD8 T lymphocytes of infected subjects express higher level of p53 tumor suppressor gene. The expression of mdm2 gene, a repressor of p53 whose increased expression is associated with apoptosis, was increased only in CD8 T cells.

CONCLUSIONS: These data confirm an activation state of CD4 and CD8 T cells in HIV-infected patients. However, the immune activation is associated with active cycling only in CD8 T cells while the pattern of gene expression observed in CD4 cells is rather in favour of a selective cell cycle arrest.

Keywords: AEGIS, CD8-Positive T-Lymphocytes, Antigens, CD4, HIV-1, CD4-Positive T-Lymphocytes, Antigens, CD8, HIV Infections, Antigens, CD28, Antigens, CD3, In Vitro, Human, analysis, immunology

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WeOrA1344