HIV IN SEMEN-EFFECTS OF STDS AND ANTIVIRAL THERAPY

描述（由申请人提供）：在过去的10年里，我们一直专注于精液中HIV的生物学。最近，我们使用了一种新的HIV RNA检测策略来识别急性HIV感染者。我们发现，在我们马拉维性病诊所的患者中，急性艾滋病毒感染非常频繁（>2%的所有客户，艾滋病，2004年）。这类患者在精液中排出大量艾滋病毒，因此极有可能发生二次性传播。这一更新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。在目标II中，我们将研究血液和精液中HIV复制的动力学，以检验精液中HIV浓度比血液中下降得更快且水平更低的假设。我们将检查传播时精浆中HIV病毒多样性的程度，并监测病毒随时间的变化。为此，我们将使用异源双链追踪试验（HTA）。我们将使用荧光原位杂交（FISH）来确定受感染的精液细胞的数量，以及每个细胞中HIV变体的数量。我们将确定性伴侣的感染状况，并比较一致性伴侣的HIV变异。在目的III中，我们将研究急性感染患者的时间CTL应答。这种反应应该预测随着时间的推移病毒负荷的大小，以及病毒突变的情况。我们将研究HIV阳性和阴性性伴侣的CTL反应。我们将使用ELISPOT分析来检查精液CDS细胞中的CTL应答。我们将针对艾滋病毒阴性伴侣开展预防工作，因为他们感染艾滋病毒的风险很大。我们相信，本申请中提出的研究将为急性HIV感染和HIV传播的生物学提供独特的见解，并为生物学和行为预防策略提供关键信息。

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.