RESUMO
In order to identify cellular genes which interfere with HIV-1 replication in monocyte-derived macrophages (MAC), cells were stimulated with interferon (IFN) or lipopolysaccharide (LPS) leading to a pronounced inhibition of HIV-1 infection in these cells, and the resulting gene expression was analyzed. Using the microarray technology we identified a gene named Stimulated Trans-Acting Factor of 50 kDa (Staf50), which is known to repress the activity of the HIV-1 LTR. Analysis of the Staf50 expression by real-time PCR showed an overexpression in IFNalpha (up to 20-fold) and LPS (up to 10-fold)-stimulated MAC as well as in infected cells (up to 3-fold). For stable overexpression, 293 T cells and primary macrophages were transduced with Staf50-IRES-GFP bicistronic pseudotype viruses. After transduction, 293 T CD4/CCR5 and MAC were infected with HIV-1, and virus replication was monitored by p24 ELISA. Overexpression of Staf50 inhibited the HIV-1 infection between 50% and 90% in 293 T CD4/CCR5 as well as in MAC. Our findings suggest that host genetic effects in combination with viral properties determine the susceptibility of an appropriate target cell for HIV-1 infection as well as the replication potential of the virus in the cell resulting in an overall productive infection.
Assuntos
HIV-1/fisiologia , Macrófagos/virologia , Proteínas Repressoras/metabolismo , Proteínas Repressoras/farmacologia , Fatores de Transcrição/metabolismo , Fatores de Transcrição/farmacologia , Regulação para Cima , Replicação Viral/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Perfilação da Expressão Gênica , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , Humanos , Interferon-alfa/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/imunologia , Macrófagos/metabolismo , Antígenos de Histocompatibilidade Menor , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Proteínas com Motivo TripartidoRESUMO
We previously showed that T cells expressing granzyme (gzm) A are more frequent in skin lesions of susceptible mice than in those of resistant mice infected with the intracellular parasite Leishmania major. To determine the in vivo role of gzm in cutaneous leishmaniasis, we examined the course of L. major infection in gzmA-deficient mice. Despite a delay in host colonization of susceptible mice, the lack of gzmA did not influence the course of lesion development or result in a discernible alteration of the interferon-gamma and interleukin-4 production. Moreover, no differences in these parameters were observed between wild-type controls and mice deficient in gzmB or both gzmA and gzmB. These findings indicate that neither gzmA nor gzmB are critical for the development of T helper cell responses and the outcome of L. major infection.