Session 7Oral Abstract Presentations Immune Responses to HIV Session Day and Time: Tuesday 10 am - 12:15 pm Presentation Time: 11:45 Room: Ballroom B
34 Systematic Microarray Analysis Reveals Counter Balance between Perforin and Interleukin 7 Receptor with Progression of HIV Infection F. Boutboul1, D. Puthier2, C. Nguyen2, H. Ait Mohand3, C. Katlama3, G. Carcelain1, P.Debré1, I. Hirsch4, B.Autran*1 1Immunol Cell, Univ Pitié Salpétrière, Paris, France; 2INSERM ERM 206, Marseille, France; 3Hosp Pitié Salpétrière, Paris, France; and 4INSERM U372, Marseille, France
Background: The use of cDNA microarray technology allows the simultaneous analysis of mRNA expression levels of thousands of genes. We used this new technology to obtain a more comprehensive view of the global effects of HIV infection on immune system cells.
Methods: We performed nylon microarrays including over 2,500 human genes to compare the kinetics of gene expression in peripheral blood mononuclear cells (PBMCs) stimulated in vitro with CD3/CD28 from 8 healthy donors and in PBMCs from 27 HIV-infected patients (pts) at different stages of the disease. Complex probes were prepared from total RNA by simultaneous reverse transcription and 33P dCTP labeling. After data processing, analysis was performed by hierarchical clustering. A multi-parameter flow cytometry analysis was also done.
Results : Despite finding a clear early-activated PBMC signature in healthy donors samples, the vast majority of these genes displays no expression variation during disease progression. Nevertheless, we identified 2 clusters of genes whose expression levels altered in response to CD3/CD28 stimulation of PBMCs in healthy donors and during HIV disease progression. The former includes 9 genes in which expression levels increase in activated PBMCs from healthy donors and from PBMCs of HIV-infected pts with uncontrolled viral load. This cluster of genes includes perforin and Type I interferon-induced proteins. The 2nd cluster of genes includes 5 genes whose expression levels decrease in activated PBMCs from healthy donors and in HIV-infected pts PBMCs with uncontrolled viral load. This cluster of genes includes interleukin 7 receptor and genes involved in TCR signaling. These findings allowed us to describe a functional link between up-regulation of perforin and down-regulation of interleukin 7 receptor on normal activated CD8 T-cells and on CD8 T-cells from HIV-infected pts.
Conclusions: Systematic microarrays analysis allows global analysis of mechanisms of immune cells alterations during HIV infection and highlights new aspects so far misunderstood of CD8 T-cell homeostasis.
