Pananghat Ambili Nair, Aggarwal Heena, Prakash Somi Sankaran, Makhdoomi Muzamil Ashraf, Singh Ravinder, Lodha Rakesh, Ali Shakir, Srinivas Maddur, Das Bimal Kumar, Pandey Ravindra Mohan, Kabra Sushil Kumar, Luthra Kalpana
From the Department of Biochemistry (ANP, HA, SSP, MAM, KL), Department of Pediatrics (RS, RL, SKK), Department of Microbiology (BKD), Department of Pediatrics Surgery (MS), Department of Biostatistics (RMP), All India Institute of Medical Sciences (RMP), and Department of Biochemistry, Jamia Hamdard University, New Delhi, India (ANP, SA).
Medicine (Baltimore). 2016 May;95(21):e3734. doi: 10.1097/MD.0000000000003734.
Disease progression in HIV-1 infected children is faster than in adults. Less than 5% of the infected children maintain stable CD4 counts beyond 7 years of infection and are termed long-term nonprogressors (LTNPs). Delineating the host immune response in antiretroviral naïve (ART) and treated HIV-1 infected children at different disease stages will help in understanding the immunopathogenesis of the disease.A total of 79 asymptomatic, perinatally HIV-1 infected children (50 ART naïve and 29 ART treated) and 8 seronegative donors were recruited in this study. T- and B-cell activation PCR arrays were performed from the cDNA, using total RNA extracted from the peripheral blood mononuclear cells (PBMCs) of 14 HIV-1 infected children at different stages of the disease. The differentially expressed genes were identified. Quantitative RT-PCR was performed for the (interleukin-8) IL-8 gene and its transcriptional mediators, that is, SHP2, GRB2, and IL-8R (IL-8 receptor/CXCR1). Plasma levels of IL-8 were measured by flow cytometry.Gene array data revealed a higher expression of IL-8 in the ART naïve HIV-1 infected progressors and in ART nonresponders than LTNPs and ART responders, respectively. Quantitative RT-PCR analysis demonstrated a significant higher expression of IL-8 (P < 0.001), its receptor CXCR1 (P = 0.03) and the upstream signaling molecule SHP2 (P = 0.04) in the progressors versus LTNPs. Plasma levels of IL-8 were significantly higher in progressors versus LTNPs (P < 0.001), and ART nonresponders versus ART responders (P < 0.001). A significant negative correlation of plasma levels of IL-8 with CD4 counts (cells/μL) was observed in HIV-1 infected ART naïve subjects (r = -0.488; P < 0.001), while the IL-8 levels positively correlated with viral load in the ART treated children (r = 0.5494; P < 0.001). ART naïve progressors on follow up demonstrated a significant reduction in the mRNA expression (P = 0.05) and plasma levels of IL-8 (P = 0.05) post 6 months of ART initiation suggesting the beneficial role of ART therapy in reducing inflammation in infected children.Our data suggest that IL-8 may serve as a potential prognostic marker in adjunct with CD4 counts to monitor disease progression in the HIV-1 infected children and the efficacy of ART.
HIV-1感染儿童的疾病进展比成人更快。不到5%的感染儿童在感染7年后CD4细胞计数保持稳定,被称为长期无进展者(LTNP)。描绘不同疾病阶段未接受抗逆转录病毒治疗(ART)和接受ART治疗的HIV-1感染儿童的宿主免疫反应,将有助于理解该疾病的免疫发病机制。本研究共招募了79名无症状的围产期HIV-1感染儿童(50名未接受ART治疗,29名接受ART治疗)以及8名血清阴性供者。使用从14名处于疾病不同阶段的HIV-1感染儿童外周血单个核细胞(PBMC)中提取的总RNA,从cDNA进行T细胞和B细胞激活PCR阵列分析。鉴定出差异表达基因。对白细胞介素-8(IL-8)基因及其转录调节因子即SHP2、GRB2和IL-8R(IL-8受体/CXCR1)进行定量逆转录PCR。通过流式细胞术测量血浆IL-8水平。基因阵列数据显示,未接受ART治疗的HIV-1感染进展者和ART无反应者中IL-8的表达分别高于LTNP和ART有反应者。定量逆转录PCR分析表明,进展者中IL-8(P<0.001)、其受体CXCR1(P=0.03)和上游信号分子SHP2(P=0.04)的表达显著高于LTNP。进展者的血浆IL-8水平显著高于LTNP(P<0.001),ART无反应者显著高于ART有反应者(P<0.001)。在未接受ART治疗的HIV-1感染受试者中,观察到血浆IL-8水平与CD4细胞计数(细胞/μL)呈显著负相关(r=-0.488;P<0.001),而在接受ART治疗的儿童中,IL-8水平与病毒载量呈正相关(r=0.5494;P<0.001)。随访中,未接受ART治疗的进展者在开始ART治疗6个月后,IL-8的mRNA表达(P=0.05)和血浆水平(P=0.05)显著降低,表明ART治疗对减轻感染儿童炎症具有有益作用。我们的数据表明,IL-8可能作为一种潜在的预后标志物,与CD4细胞计数一起用于监测HIV-1感染儿童的疾病进展和ART的疗效。