HLA-G VARIANT ASSESSMENT IN VERTICAL TRANSMISSION OF HIV

彻底了解影响宿主-HIV-1相互作用的致病性和传播后果的遗传因素（宿主和病毒）对公共卫生的诊断、传播和预防策略的各个方面都至关重要。具体目的：探讨HLA-G等位基因在垂直传播中的潜在作用。目的：i）筛选、鉴定和表征来自HIV-1感染和未感染的母亲/儿童对的HLA-G DNA样品中的遗传变异，并比较样品之间的HLA-G百分比变异性。ii）评估外显子2-7中HLA-G等位基因序列变异频率与母亲/子女对之间垂直传播的关系之间的潜在相关性。方法：将从CDC获得HIV-1感染和未感染母亲/儿童对的DNA样本（PAPER-研究）。本研究将采用RAPD-PCR、等位基因特异性PCR、SSCP-PCR，然后进行DNA克隆/测序。将切下所有不同的PCR条带并克隆。将对每个PCR产物约5-10个克隆进行测序。将使用PASA方法确认可能存在的任何突变。Fisher精确检验（x2）、逻辑回归将用于比较组值。将使用最大似然、Hardy-Weinberg平衡和连锁不平衡参数分析来检查样本中每个基因座的任何HLA-G突变的随机分布，以及HLA-G变异与HIV-1垂直传播频率的风险相关性。

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.