Macrophage-derived chemokine gene expression in human and macaque cells: mRNA quantification using NASBA technology

Macrophage-derived chemokine (MDC) is a CC-chemokine that inhibits infectio n by both macrophage- and T cell-tropic strains of HIV-1, This suppressor a ctivity has led to great interest in fully characterizing the role of MDC i n the pathogenesis of HIV-1 infection. Methods for the quantitation of cons titutive levels of MDC protein in vivo are lacking. In this report, we desc ribe the development and performance of a NASBA-based assay for the quantif ication of MDC mRNA expression in human and macaque cells. Although the con stitutive in vivo levels of MDC mRNA in macaque and human T lymphocytes wer e low, in vitro activation of these cells greatly increased MDC transcripti on. Levels in the human and macaque cells were comparable under all conditi ons tested. Positive correlations between MDC transcription and protein exp ression were observed. The results indicate that this assay is extremely se nsitive and reproducible over a five log dynamic range, and effectively qua ntifies MDC mRNA in resting and activated T cells. This assay may therefore permit characterization of the role of MDC in HIV-1/SIV pathogenesis, and in vaccine-induced immune responses. (C) 2001 Academic Press.