目的定量研究HIV-1感染者外周血单个核细胞(PBMC)中胞嘧啶脱氨基酶家族成员hA3B、hA3F和hA3G的mRNA表达水平,并分析它们与CD4^+T淋巴细胞计数的相关性。方法采集各21例未接受和已接受抗病毒治疗的HIV-1感染者、10例HIV-1阴性健康对照者的外周静脉血,Ficoll密度梯度离心法分离PBMC,采用反转录-实时荧光定量PCR法检测其中hA3B、hA3F和hA3GmRNA水平,流式细胞术检测CD4^+T淋巴细胞计数。数据行t、t'或Wilcoxon秩和检验。结果未治疗及已治疗的HIV-1感染者、HIV-1阴性者hA3BmRNA的表达水平分别为208.4、365.2和563.6(P〈0.05),hA3FmRNA分别为245.5、316.6和442.9(P〈0.05),hA3GmRNA分别为404.6、360.8和638.6。HIV-1感染者hA3GmRNA水平低于健康对照(Z=2.451,Z=2.493,均P〈0.05),但未治疗与已治疗HIV-1组间比较差异无统计学意义(Z=0.340,P=0.7342)。HIV-1感染者hA3B、hA3F和hA3GmRNA表达水平与CD4^+T淋巴细胞计数均无相关性(未治疗者:r=-0.0104,r=-0.0545,r=0.1623,均P〉0.05;已治疗者:r=0.3220,r=0.2193,r=0.1455,均P〉0.05)。hA3B、hA3F和hA3G三者mRNA水平在已治疗HIV-1感染者和健康对照者中呈正相关(P〈0.05),而在未治疗HIV-1感染者无相关性(P〉0.05)。结论hA3B、hA3F和hA3G基因表达水平与CD4^+T淋巴细胞计数无直接相关性;HIV-1感染可导致PBMC中hA3B、hA3F和hA3G表达水平降低,抗病毒治疗可上调hA3B和hA3F的表达。
Objective To quantitatively investigate the expression levels of the cytosine deaminases, hA3B, hA3F and hA3G in peripheral blood mononuclear cell (PBMC) of human immunodeficiency virus (HIV) infected patients and to analyze the correlation between cytosine deaminases expression and CD4^+ T lymphocyte counts. Methods Peripheral blood samples were collected from 21 HIV-infected subjects who didn't take antiretroviral therapy (ART-), 21 HIV- infected subiects receiving ART(ART+), and 10 HIV uninfected subjects. PBMC were isolated by Ficoll density gradient eentrifugation, followed by RNA extraction and cDNA synthesis, hA3B, hA3F and hA3G mRNA levels were determined by real-time fluorescent quantitative polymerase chain reaction (PCR). CD4^+ T lymphocyte counts were determined using flow cytometry. Data were analyzed by t test,t' test or Wilcoxon rank sum test. Results In HIV-infected subjects without or with ART, HIV-uninfected subjects , the levels of hA3B mRNA were 208.4, 365.2 and 563.6, hA3F mRNA were 245.5, 316. 6 and 442.9, hA3G mRNA were 404. 6, 360.8 and 638. 6, respectively. hA3G mRNA level in HIV-infected subjects was lower than that in HIV-uninfected controls (P = 0. 0131), but there was no statistical difference between ART+ and ART- groups(P=0. 7342). There were no correlations between hA3B, hA3F and hA3G mRNA levels and CD4^+ T lymphocyte counts in either ART-- or ART+ HIV-infected subjects(ART-- : r= -0. 0104,r= -0. 0545,r=0. 1623, all P〉0.05; ART+ : r= 0. 3220, r= 0. 2193, r= 0. 1455, all P〉0. 05). In addition, hA3B, hA3F and hA3G mRNA levels were positively correlated with one another in ART- HIV-infected subjects and HIV uninfected controls (P〈0.05), but not in ART-- HIV-infected subjects (P〉0.05). Conclusions hA3B, hA3F and hA3G expression levels do not directly correlate with CD4^+ T lymphocyte counts in HIV 1-infected patients, hA3B, hA3F and hA3G expression levels in PBMCs tend to be decreased after HIV-1 infection, and A