Munshi Saif Ullah, Panda Harekrushna, Holla Prasida, Rewari Bharat Bhushan, Jameel Shahid
Virology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
ICGEB-Emory Vaccine Centre, International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
PLoS One. 2014 May 14;9(5):e95920. doi: 10.1371/journal.pone.0095920. eCollection 2014.
The surrogate markers of HIV/AIDS progression include CD4 T cell count and plasma viral load. But, their reliability has been questioned in patients on anti-retroviral therapy (ART). Five microRNAs (miRNAs) - miR-16, miR-146b-5p, miR-150, miR-191 and miR-223 in peripheral blood mononuclear cells (PBMCs) were earlier found to assign HIV/AIDS patients into groups with varying CD4 T cell counts and viral loads. In this pilot study, we profiled the expression of these five miRNAs in PBMCs, and two of these miRNAs (miR-146b-5p and miR-150) in the plasma of HIV/AIDS patients, including those on ART and those who developed ART resistance, to evaluate if these are biomarkers of disease progression and therapy.
We quantified miRNA levels by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using RNA isolated from PBMCs and plasma of healthy persons or HIV-infected patients who were (1) asymptomatic; (2) symptomatic and ART naïve; (3) on ART; and (4) failing ART. Our results show miR-150 (p<0.01) and to a lesser extent miR-146b-5p (p<0.05) levels in PBMCs to reliably distinguish between ART-naïve AIDS patients, those on ART, and those developing drug resistance and failing ART. The plasma levels of these two miRNAs also varied significantly between patients in these groups and between patients and healthy controls (p values <0.05).
We report for the first time that PBMC and plasma levels of miR-150 and miR-146b-5p are predictive of HIV/AIDS disease progression and therapy.
HIV/AIDS进展的替代标志物包括CD4 T细胞计数和血浆病毒载量。但是,它们在接受抗逆转录病毒治疗(ART)的患者中的可靠性受到了质疑。外周血单核细胞(PBMCs)中的5种微小RNA(miRNA)——miR-16、miR-146b-5p、miR-150、miR-191和miR-223,早期被发现可将HIV/AIDS患者分为具有不同CD4 T细胞计数和病毒载量的组。在这项初步研究中,我们分析了这5种miRNA在PBMCs中的表达情况,以及其中两种miRNA(miR-146b-5p和miR-150)在HIV/AIDS患者血浆中的表达情况,这些患者包括接受ART治疗的患者以及出现ART耐药的患者,以评估它们是否为疾病进展和治疗的生物标志物。
我们使用从健康人或HIV感染患者的PBMCs和血浆中分离的RNA,通过定量逆转录聚合酶链反应(qRT-PCR)对miRNA水平进行了定量分析,这些患者包括:(1)无症状的;(2)有症状且未接受ART治疗的;(3)正在接受ART治疗的;(4)ART治疗失败的。我们的结果显示,PBMCs中miR-150(p<0.01)以及程度稍轻的miR-146b-5p(p<0.05)水平能够可靠地区分未接受ART治疗的AIDS患者、接受ART治疗的患者以及出现耐药和ART治疗失败的患者。这两种miRNA的血浆水平在这些组的患者之间以及患者与健康对照之间也有显著差异(p值<0.05)。
我们首次报告,PBMCs和血浆中miR-150和miR-146b-5p的水平可预测HIV/AIDS疾病的进展和治疗情况。