The gender specific prevalence of multiple strain genital HSV-2 infection

描述（由申请人提供）：生殖器单纯疱疹病毒2型（HSV-2）是一种性传播感染，主要影响女性。在美国，女性的2型单纯疱疹病毒血清患病率几乎是男性的两倍，而在世界范围内，感染2型单纯疱疹病毒的女性比男性多近1亿。在撒哈拉以南非洲，2型单纯疱疹病毒感染是艾滋病毒流行的主要驱动因素之一。虽然针对2型单纯疱疹病毒的疫苗将极大地有利于妇女的健康，但迄今为止，疫苗开发的尝试都失败了。尽管在动物模型中取得了成功，并在人类中诱导了强有力的免疫反应，但基于糖蛋白的疫苗并不能预防HSV-2的获得。最近的小型研究描述了多株HSV-2感染，但其流行程度和危险因素尚不清楚。重要的是，如果多株感染是常见的，这将表明疫苗将需要产生比自然感染产生的免疫反应更有效的免疫反应。我们的目标是确定是否以及在多大程度上发生多株2型单纯疱疹病毒感染，因为这将指导新疫苗是否需要引发“比自然更好”的免疫。我们的具体目的是确定多株HSV-2感染的性别特异性患病率和危险因素。我们假设：1)女性比男性更容易感染多株HSV-2生殖器感染；2)终生性伴侣数量多的人比终生性伴侣数量少的人更容易感染生殖器HSV-2；3)与美国女性相比，来自撒哈拉以南非洲的女性将有更高的多菌株感染患病率；4)与HIV血清阴性者相比，HIV血清阳性者多株感染的患病率更高。在我们广泛的病毒学和流行病学研究中，我们前瞻性地从bb800名参与者中收集了一系列独特的HSV-2 DNA阳性生殖器样本。这些样本包括来自美国和撒哈拉以南非洲地区的男性和女性，以及来自秘鲁的男性。HIV血清阳性的人包括在美国和非洲的样本中。所有参与者均有生殖器HSV-2感染的临床和病毒学特征，并提供详细的性史。获得这些标本使我们的小组处于一个独特的位置来解决这些假设。我们的技术方法是使用下一代测序（NGS）在48个生殖器拭子样本中鉴定高变区和流行snp，然后使用鉴定的区域对来自600名参与者的标本集使用焦磷酸测序进行高通量，多位点基因分型，以准确表征每个人体内标本集的单株与多株感染。总之，这些数据将澄清性别与2型单纯疱疹病毒易感性之间的关系，并为2型单纯疱疹病毒疫苗的可能疗效提供基准。

DESCRIPTION (provided by applicant): Genital herpes simplex virus type 2 (HSV-2) is a sexually transmitted infection that disproportionately affects women. HSV-2 seroprevalence is nearly two-fold higher in women than in men in the US, and worldwide, nearly 100 million more women than men are HSV-2 infected. In sub-Saharan Africa, HSV-2 infection is one of the main drivers of the HIV epidemic. While a vaccine against HSV-2 would greatly benefit women's health, attempts at vaccine development have failed thus far. Despite success in animal models and induction of potent immune responses in humans, the glycoprotein based vaccines do not protect against HSV-2 acquisition. Recent small studies have described multiple-strain HSV-2 infection but the prevalence and risk factors for this are not known. Importantly, if multiple-stran infection is common, it will suggest that vaccines will need to produce immune responses that are more potent than those produced in natural infection. Our goal is to define whether, and to what extent, multiple-strain HSV-2 infection occurs, as that will guide whether new vaccines need to elicit "better than nature" immunity. Our Specific Aim is to determine the gender- specific prevalence of and risk factors for multiple-strain HSV-2 infection. We hypothesize that 1) women will be more likely than men to have genital HSV-2 infection with multiple strains; 2) persons with higher numbers of lifetime sex partner will be more likely to have genital HSV-2 that those with fewer partners; 3) Women from sub-Saharan Africa will have higher prevalence of multiple strain infection as compared with U.S. women; 4) HIV seropositive persons will have a higher prevalence of multiple strain infection as compared with HIV seronegative persons. We have prospectively collected unique sets of serial HSV-2 DNA positive genital samples from >800 participants enrolled in our extensive virologic and epidemiologic studies. These include samples from women and men from the US and sub-Saharan Africa, and men from Peru. HIV seropositive persons are included both in the US and African samples. All participants have clinically and virologically characterized genital HSV-2 infection, and provided detailed sexual histories. Access to these specimens places our group in a unique position to address these hypotheses. Our technical approach is to use Next-Generation Sequencing (NGS) to identify hypervariable regions and prevalent SNPs in 48 genital swab samples, and then use identified regions to perform high-throughput, multi-locus genotyping using pyrosequencing on specimen sets from 600 participants, to accurately characterize each within-person specimen set as single vs. multiple strain infection. Together, these data will clarify the relationship between gender an HSV-2 susceptibility and provide a benchmark for the likely efficacy of HSV-2 vaccines.