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，不成比例地影响妇女。在美国，HSV-2血清阳性率在女性中比男性高近两倍，在全世界，HSV-2感染的女性比男性多近1亿。在撒哈拉以南非洲，HSV-2感染是艾滋病毒流行的主要驱动因素之一。虽然针对HSV-2的疫苗将极大地有益于妇女的健康，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有效的免疫应答，但基于糖蛋白的疫苗不能防止HSV-2的获得。最近的小型研究描述了多株HSV-2感染，但其患病率和风险因素尚不清楚。重要的是，如果多链感染很常见，这将表明疫苗需要产生比自然感染更有效的免疫反应。我们的目标是确定是否以及在多大程度上发生多株HSV-2感染，因为这将指导新疫苗是否需要引起“优于自然”的免疫力。我们的具体目标是确定多株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生殖器HSV-2多株病毒; 2）终身性伴侣数量较多的人比伴侣数量较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多株病毒感染的患病率更高; 4）艾滋病病毒血清反应阳性的人比艾滋病病毒血清反应阴性的人有更高的多株感染率。我们从800多名参与我们广泛的病毒学和流行病学研究的参与者中前瞻性地收集了独特的系列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的女性和男性，以及来自秘鲁的男性。艾滋病毒血清反应阳性者包括在美国和非洲的样本中。所有参与者都有生殖器HSV-2感染的临床和病毒学特征，并提供了详细的性史。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）来识别48个生殖器拭子样本中的高变区和流行SNP，然后使用识别的区域对来自600名参与者的样本集使用焦磷酸测序进行高通量、多位点基因分型，以准确地将每个人内样本集表征为单一与多菌株感染。总之，这些数据将阐明性别与HSV-2易感性之间的关系，并为HSV-2疫苗的可能有效性提供基准。 公共卫生相关性：作为HSV-2感染自然史研究的一部分采集的生殖器拭子将使用PCR扩增和测序进行HSV-2菌株分型，以确定1）生殖器是否经常发生HSV-2多种菌株感染，2）与男性相比，这在女性中是否更常见，3）阐明多种菌株感染的风险因素，如大量终身性伴侣，HIV感染，以及4）如果多种菌株感染的流行因地理位置而异。

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. PUBLIC HEALTH RELEVANCE: Genital swabs collected as part of natural history studies of HSV-2 infection will undergo HSV-2 strain typing using PCR amplification and sequencing to determine 1) if genital infection with multiple strains of HSV-2 occurs frequently, 2) if this is mre common in women as compared to men and 3) to elucidate risk factors for multiple strain infection, such as large number of lifetime sexual partners, HIV infection, and 4) if the prevalence of multiple strain infection varies by geographic location.