HLA-G VARIANT ASSESSMENT IN VERTICAL TRANSMISSION OF HIV

彻底了解影响宿主- hiv -1相互作用的致病和传播后果的遗传因素（包括宿主和病毒）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：研究HLA-G等位基因在垂直传播中的潜在作用。目的：i)筛选、鉴定和描述来自hiv -1感染和未感染母亲/儿童对的HLA-G DNA样本的遗传变异，并比较样本之间HLA-G百分比变异性。ii)评估2-7外显子HLA-G等位基因序列变异频率与母婴间垂直传播的潜在相关性。方法：hiv -1感染和未感染的母亲/儿童对的DNA样本将从CDC （PACTS-研究）获得。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。所有不同的PCR条带将被切除并克隆。每个PCR产物大约5-10个克隆将被测序。PASA方法将用于确认可能存在的任何突变。采用Fisher精确检验（x2）、logistic回归进行组值比较。最大似然、Hardy-Weinberg平衡和连锁不平衡参数分析将用于检查样本中每个位点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.