HLA-G VARIANT ASSESSMENT IN VERTICAL TRANSMISSION OF HIV

彻底了解影响宿主-HIV-1相互作用致病和传播后果的遗传因素(包括宿主和病毒)，对于从诊断、传播和预防策略的公共卫生的各个方面都是至关重要的。具体目的：研究人类白细胞抗原G等位基因在垂直传播中的潜在作用。目的：i)筛查和鉴定HIV-1感染和未感染母婴对的人类白细胞抗原-G DNA样本中的基因变异，并比较样本之间的人类白细胞抗原-G百分比变异。Ii)评估人类白细胞抗原-G等位基因序列外显子2-7变异频率与母子间垂直传播关系的潜在相关性(S)。方法：HIV-1感染和未感染母婴的DNA样本将从疾控中心获得(PACTS-Study)。本研究将采用随机扩增、等位基因特异性聚合酶链式反应、单链构象多态性聚合酶链式反应，然后进行DNA克隆和测序。所有不同的聚合酶链式反应条带都将被切除和克隆。每个PCR产物大约5-10个克隆将被测序。将使用PASA方法来确认可能存在的任何突变。组间比较采用Fisher‘s精确检验(X2)、Logistic回归分析。采用最大似然法、Hardy-Weinberg均衡法和连锁不平衡参数法检验样本中各位点HLAG突变的随机分布，以及HLAG变异(S)对HIV-1垂直传播频率的风险关联。

A thorough understanding of genetic factors (both host and viral) effecting the pathogenic and transmission consequences of host-HIV-1 interactions is essential to all aspects of public health from diagnostic, transmission, and prevention strategies. Specific aim: The potential role played by HLA-G allele in vertical transmission will be examined. Aims: i) Screen and identify and characterize genetic variations in the HLA-G DNA samples from HIV-1-infected and uninfected mother/child pairs and compare HLA-G percentage variability among samples. ii) Evaluate for potential correlation(s) between HLA-G allele sequence variant frequencies in exons 2-7 and relationships to vertical transmission among the mother/child pairs. Approach: DNA samples of HIV-1-infected and uninfected mother/child pairs will be obtained from CDC (PACTS- study). RAPD-PCR, allele-specific PCR, SSCP-PCR will be employed in this study, followed by DNA cloning/sequencing. All distinct PCR bands will be excised and cloned. Approximately 5-10 clones per PCR product will be sequenced. PASA method will be used to confirm any mutation that may exist. Fisher's exact test (x2), logistic regression will be utilized to compared the groups value. Maximum likelihood, Hardy-Weinberg equilibrium, and linkage disequilibrium parameter analyses will be used to check for random distribution of any mutation of HLA-G at each locus among the samples, and the risk association of HLA-G variation(s) on the frequency of HIV-1 vertical transmission.