Serial Analysis of Gene Expression in CD8+ T-cells isolated from HTLV-1 infected individuals / Análise do perfil de expressão gênica em linfócitos T CD8+ isolados de pacientes infectados pelo HTLV-1

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

MALTA, T.M. Serial Analysis of Gene Expression in CD8+ T-cells isolated from HTLV-1 infected individuals. 2009. 137f. Dissertação de Mestrado. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, Ribeirão Preto, 2009. Human T-cell lymphotropic virus type 1 (HTLV-1) was the first human retrovirus discovered and is related with two major diseases: adult T cell lymphoma/leukaemia (ATLL) and HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). About 3 to 5% of infected individuals will develop HTLV-1 related diseases, while the majority remains life-long asymptomatic carriers of the virus. HAM/TSP is an inflammatory manifestation of central nervous system and the mechanism involved in HAM/TSP development are not well elucidated. CD8+ cytotoxic T lymphocytes (CTL) have an important function in the immune response against the HTLV-1 and is related with the individual proviral load and so in the risk of HAM/TSP. To identify genes differentially expressed among non-infected individuals, asymptomatic (HAC) and HAM/TSP (HAM/TSP) patients we performed a serial analysis of gene expression (SAGE) using CD8+ T cells isolated from these individuals. The CD8+ T lymphocytes were isolated by magnetic cell separation system and HAC and HAM/TSP SAGE libraries were composed by pooled of four samples. SAGE analysis of 51,017, 62,432 and 60,620 tags from control, HAC and HAM/TSP groups respectively, allowed identification of approximately 12,000 different transcripts in each library. The expression profile revealed around 900 genes differentially expressed between control group and HAC or HAM/TSP, and 290 genes were identified between HAC and HAM/TSP groups. The expression profile revealed the presence of highly frequent transcripts related to apoptosis, cell adhesion and cell migration. These results were validated by real time PCR in 17 HTLV-1 asymptomatic carriers, 14 patients with HAM/TSP and 24 healthy individuals. The validation revealed the increased levels of expression of the cytolysis genes PRF1, GZMB and GZMH, and the increases of CCL5, ZAP70 and PXN in the HTLV-1 infected individuals. Besides, the CXCR4 was suppressed in these patients. Additionally, we performed the protein quantification of PRF1 and GZMB by flow cytometry and the real time PCR results were confirmed. The CD8+ T cells profiles showed strong cell activation and cell migration in HTLV-1 infection. These data suggests that CD8+ T cells from HTLV-1 individuals seem to contribute more to the protection and virus infection control than to the HAM/TSP development. This study represents the first extensive serial analysis of gene expression of CD8+T cells in HTLV-1 infection and allowed to generate data that will be used in different approaches in HTLV-1 research.

ASSUNTO(S)

sage htlv-1 ham/tsp ham/tsp expressão gênica. htlv-1 gene expression. sage

ACESSO AO ARTIGO

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