HIV IN SEMEN-EFFECTS OF STDS AND ANTIVIRAL THERAPY

描述(申请人提供)：在过去的10年里，我们一直专注于精液中艾滋病毒的生物学研究。最近，我们使用了一种新的艾滋病毒RNA检测策略来识别急性艾滋病毒感染者。我们发现，在马拉维的性病诊所，急性艾滋病毒感染在患者中非常频繁(2004年，艾滋病，占所有客户的2%)。这些患者在精液中排泄了如此多的艾滋病毒，以至于二次性传播变得非常有可能。这份续签申请包括对马拉维急性艾滋病毒感染的详细研究。在AIM I中，我们将招募200多名患有急性艾滋病毒感染的男性和女性及其一些性伴侣的纵向队列。我们将评估简化检测策略识别急性艾滋病毒感染者的能力。在AIM II中，我们将研究血液和精液中艾滋病毒复制的动态，以检验精液中艾滋病毒浓度下降更快且在精液中的水平比血液中更低的假设。我们将检测传播时精浆中艾滋病毒病毒的多样性程度，并监测病毒随时间的变化(S)。为此，我们将使用异源双链跟踪试验(HTA)。我们将使用荧光原位杂交(FISH)来确定感染的精细胞数量，以及每个细胞中HIV变异的数量。我们将确定性伴侣的感染状况，并比较性伴侣中HIV变异的情况。在AIM III中，我们将检测急性感染患者的颞叶CTL反应。这一反应应该可以预测随着时间的推移病毒负担的大小，以及病毒突变的场景(S)。我们将检查HIV阳性和阴性性伴侣的CTL反应。我们将使用ELISPOT试验来检测精液CDS细胞的CTL反应。我们将针对艾滋病毒阴性的伴侣开展预防工作，他们有相当大的感染艾滋病毒的风险。我们相信，本申请中提出的研究将为急性艾滋病毒感染和艾滋病毒传播的生物学提供独特的见解，并提供对生物和行为预防战略至关重要的信息。

DESCRIPTION (provided by applicant): For the past 10 years we have focused on the biology of HIV in semen. Most recently we have used a novel HIV RNA detection strategy to identify people with acute HIV infection. We found that acute HIV infections are remarkably frequent in patients in our STD clinic in Malawi (>2% of all clients, AIDS, 2004). Such patients excrete so much HIV in semen that secondary sexual transmission becomes extremely likely. This renewal Application includes detailed studies of acute HIV infection in Malawi. In Aim I we will recruit a longitudinal cohort of more than 200 men and women with acute HIV infection, and some of their sexual partners. We will evaluate the ability of simplified testing strategies to identify people with acute HIV infection. In Aim II we will examine the dynamics of HIV replication in blood and semen to test the hypothesis that the concentration of HIV in semen decreases faster and to a lower level in semen than in blood. We will examine the degree of HIV viral diversity in seminal plasma at the time of transmission, and monitor viral change(s) over time. We will use the heteroduplex tracking assay (HTA) for this purpose. We will use fluorescence in-situ hybridization (FISH) to determine the number of seminal cells infected, and the number of HIV variants per cell. We will determine the infection status of sexual partners, and compare the HIV variants in concordant partners. In Aim III we will examine the temporal CTL response in patients with acute infection. This response should predict the magnitude of viral burden over time, and the sight(s) of viral mutation. We will examine the CTL response in both HIV postive and negative sexual partners. We will use ELISPOT assays to examine the CTL response in seminal CDS cells. We will undertake prevention efforts directed at HIV negative partners, who are at considerable risk for HIV acquisition. We believe the studies proposed in this Application will provide unique insight into the biology of acute HIV infection and HIV transmission, and information critical for both biological and behavioral prevention strategies.