Sex Differences in Plasma HIV-1 RNA Concentrations

描述（由申请人提供）：多项研究证明，CD4+ T细胞计数大于300细胞/mm3时，女性血浆HIV-1 RNA浓度（VL）明显低于男性。VL的性别差异机制尚不清楚。由于大多数HIV-1复制发生在淋巴组织（LT）中，因此男性和女性LT的差异很可能解释了VL的差异。我们建议探索两种截然不同的理论——LT CD4+ T细胞上CCR5密度的性别差异和/或每个感染LT细胞产生的病毒粒子数量的性别差异——来解释女性血浆VL低于男性的原因。本研究的具体目的是：1)确定女性LT中CD4+ T细胞上的CCR5密度是否低于男性；2)确定每个有效感染淋巴结细胞中HIV-1 RNA的平均拷贝数在女性中是否低于男性。为了实现这一目标，将招募30名女性和35名男性HIV-1感染患者，他们没有接受抗逆转录病毒治疗，并且有300个CD4+ T细胞捐献腹股沟淋巴结（LN）和外周血。分解LN CD4+ T细胞上的CCR5密度将通过使用QuantiBRITE头系统的流式细胞术分析来确定。RT - PCR检测HIV-1 RNA将用于测定血浆和LN细胞群中的VL。LN切片中产生病毒的细胞的频率将通过HIV-1 RNA的原位杂交确定，并与LN细胞VL结合使用，计算每个感染细胞的HIV-1 RNA的平均拷贝数。在调整CD4+ T细胞计数、年龄和种族后，将通过线性回归分析数据，以估计性别、VL（在血浆和LN细胞中）、CCR5表达密度或lt中产生病毒的细胞频率之间的关系。将比较男性和女性有效感染LN细胞中HIV-1 RNA的平均拷贝数。二级分析将探讨T细胞活化、凋亡和性激素水平与VL、CCR5密度和每个细胞平均HIV-1 RNA拷贝数之间的关系。更好地了解VL的性别差异机制可能会导致对男性和女性采用不同的治疗方法，对接受激素治疗的HIV-1感染者采用不同的抗逆转录病毒指南，以及改进对HIV-1感染者治疗的新疗法。

DESCRIPTION (provided by applicant): Multiple studies have documented significantly lower plasma HIV-1 RNA concentrations (VL) in women compared to men at CD4+ T cell counts greater than 300 cells/mm3. Mechanisms underlying sex differences in VL are unknown. Because most HIV-1 replication occurs in lymphoid tissues (LT), it is likely that differences in LT of men and women account for differences in VL. We propose to explore two distinct theories - that sex differences in CCR5 density on LT CD4+ T cells and/or sex differences in numbers of virions produced per infected LT cell exist - to explain lower plasma VL in women compared to men. The specific aims of this study are: 1) To determine whether CCR5 density on CD4+ T cells in LT is lower in women compared to men; and 2) To determine whether the average number of HIV-1 RNA copies per productively infected lymph node cell is lower in women compared to men. To accomplish this, 30 women and 35 men with HIV-1 infection who are not receiving antiretroviral therapy and who have >300 CD4+ T cells will be recruited to donate inguinal lymph nodes (LN) and peripheral blood. CCR5 density on disaggregated LN CD4+ T cells will be determined by flow cytometric analysis using the QuantiBRITE bead system. RT PCR for HIV-1 RNA will be used to determine VL in plasma and LN cell populations. Frequencies of virus-producing cells in LN sections will be determined by in situ hybridization for HIV-1 RNA and used in conjunction with the LN cell VL to calculate the average number of copies of HIV-1 RNA per infected cell. Data will be analyzed by linear regression, after adjusting for CD4+ T cell count, age, and race, to estimate the relationship between sex, VL (in plasma and LN cells), and density of CCR5 expression or frequencies of virus-producing cells in LT. Average number of copies of HIV-1 RNA in productively infected LN cells will be compared between men and women. Secondary analyses will explore relationships between T cell activation, apoptosis, and sex hormone levels and VL, CCR5 density, and average HIV-1 RNA copy number per cell. A better understanding of mechanisms underlying sex differences in VL could lead to different therapeutic approaches in men and women, different antiretroviral guidelines for HIV-1-infected individuals undergoing hormonal therapies, and novel therapies that improve the treatment of both HIV-1 infected men and women.