RESUMO

IFN-gamma mRNA expression was evaluated in nonstimulated peripheral blood mononuclear cells (PBMC) of HIV-infected and seronegative individuals using quantitative competitive and semiquantitative RT-PCR and the sensitivity of these methods was compared. A significant correlation was found between quantitative competitive and semiquantitative RT-PCR in samples of both HIV-seronegative (P = 0.004) and HIV-infected individuals (P = 0.0004). PBMC from HIV-infected individuals presented a remarkable increase of IFN-gamma mRNA expression, as determined by both types of RT-PCR methods. Semiquantitative RT-PCR even without an internal standard is also acceptable for measuring cytokine mRNA expression, but less reliable if small amounts are quantified. Moreover, we found that increased IFN-gamma mRNA expression is independent of CD4+ cell count in AIDS-free HIV-infected patients.

Assuntos

Infecções por HIV/sangue , Interferon gama/sangue , Leucócitos Mononucleares/metabolismo , RNA Mensageiro/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome da Imunodeficiência Adquirida/sangue , Síndrome da Imunodeficiência Adquirida/imunologia , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos , Infecções por HIV/imunologia , Soronegatividade para HIV , Humanos , Estudos Longitudinais , Sensibilidade e Especificidade

RESUMO

IFN-gamma mRNA expression was evaluated in nonstimulated peripheral blood mononuclear cells (PBMC) of HIV-infected and seronegative individuals using quantitative competitive and semiquantitative RT-PCR and the sensitivity of these methods was compared. A significant correlation was found between quantitative competitive and semiquantitative RT-PCR in samples of both HIV-seronegative (P = 0.004) and HIV-infected individuals (P = 0.0004). PBMC from HIV-infected individuals presented a remarkable increase of IFN-gamma mRNA expression, as determined by both types of RT-PCR methods. Semiquantitative RT-PCR even without an internal standard is also acceptable for measuring cytokine mRNA expression, but less reliable if small amounts are quantified. Moreover, we found that increased IFN-gammamRNA expression is independent of CD4+ cell count in AIDS-free HIV-infected patients

Assuntos

Humanos , Infecções por HIV , Interferon gama , Leucócitos Mononucleares , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/metabolismo , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos , Infecções por HIV , Soronegatividade para HIV , Interferon gama , Estudos Longitudinais , RNA Mensageiro , Sensibilidade e Especificidade