TOPICAL THERAPY FOR HSV-2 INFECTION OF THE GENITAL TRACT

生殖器疱疹病毒感染是由男性或女性生殖器组织感染单纯疱疹病毒2型(HSV-2)引起的，或者由单纯疱疹病毒1型(HSV-1)感染的频率较低。这项提案将重点关注HSV-2。女性的感染可以针对唇面、阴道和宫颈。臀部皮肤的邻近区域也可能受到感染。感染可能作为主要事件发生，通常是性传播的结果。初次感染后，病毒最常进入骶神经节潜伏，并可作为反复感染的来源。初次感染通常更严重，但反复感染可能会在几年内发生，并成为传播病毒的一个重要来源。这种病毒感染的发病率极高。免疫抑制患者面临这种病毒在所有组织复制的严重风险，并可能遭受威胁生命的后遗症。目前治疗HSV-2感染的主要药物是阿昔洛韦，这是一种有效的核苷酸类似物，特别是在HSV感染细胞中被磷酸化并并入DNA中。该化合物的静脉、口服和外用制剂具有治疗作用。此外，某些以洗涤剂为基础的杀精剂已被证明对HSV具有抗病毒活性。本计划项目前三年的研究表明，C31G(C14/C16)和烷基硫酸盐杀菌剂都可以灭活HSV-2。重要的是，在小鼠阴道感染的活体模型中，十二烷基硫酸钠已被证明可预防HSV-2感染，在人类阴道异种移植模型中，十二烷基硫酸钠和C31G已被证明可灭活HSV-2并防止感染。我们在下一阶段的具体目标将包括：1)继续使用三种HSV-2生长模型系统来确定非配方和配方杀微生物剂的毒性和有效性：a)通过在猴肾上皮细胞和原代人阴道角质形成细胞中形成空斑来体外检测HSV-2；b)通过经阴道接种瑞士韦氏杂交小鼠进行体内检测；c)通过接种免疫低下小鼠生长的人阴道异种移植瘤进行体内检测；2)比较HSV-2感染的动力学和自然病史，无论是接种正常的人、人、阴道异种移植物还是表达HPV-11病毒全套基因的人阴道移植物；以及3)确定生长在裸鼠、SCID小鼠或与人淋巴网状细胞重组的SCID小鼠中的人阴道异种移植瘤的急性和亚临床HSV-2感染是否改变了作为HIV感染潜在靶点的异种移植瘤细胞的复杂性。

Genital herpes virus infection is caused by infection of male or female genital tissues by herpes simplex virus type 2 (HSV-2) or less frequently by herpes simplex virus type 1 (HSV-1). This proposal will focus on HSV-2. Infection in the female can target the labial surfaces, the vagina and the cervix. Adjacent areas of buttock skin also may be infected. Infection may occur as a primary event, usually as a result of sexual transmission. Following primary infection, virus most often enters latency in sacral ganglia and can serve as a source of recurrent infection. Primary infection is usually more severe, but recurrent infections may occur over a number of years and serve as a potent source of transmittable virus. Incidence of this virus infection is extremely high. Immunosuppressed patients are at grave risk for replication of this virus at all tissues and can suffer life-threatening sequelae. The current mainstay of therapy for HSV- 2 infection is Acyclovir, a potent nucleotide analogue specifically phosphorylated and incorporated into DNA in HSV-infected cells. Intravenous, oral and topical formulations of this compound have therapeutic benefit. In addition, certain detergent based spermicides have proven anti-virucidal activity for HSV. Studies in the previous three years of this Program Project have shown that C31G (C14/C16) and an alkyl sulfate microbicide can each inactivate HSV-2. Importantly, SDS has been shown to prevent HSV-2 infection in an in vivo model of infection in the mouse vaginal and SDS and C31G have been shown to inactivate HSV-2 and prevent infection in a human vaginal xenograft model. Our specific aims in the next phase of this grant will include: 1) continue to employ three model systems for HSV-2 growth to determine the toxicity and efficacy of non-formulated and formulated microbicidal compounds: a) in vitro assay of HSV-2 by plaque formation in monkey kidney epithelial cells and primary human vaginal keratinocytes; b) in vivo assay by vaginal inoculation of Swiss-Webster, outbred mice, c) in vivo assay by inoculation of human, vaginal xenografts growing in immunocompromised mice; 2) compare the kinetics and natural history of HSV-2 infection following inoculation of either normal, human, vaginal xenografts or human vaginal xenografts expressing the complete repertoire of viral genes from HPV-11; and 3) determine if acute of subclinical HSV-2 infection of human vaginal xenografts growing in nude mice, SCID mice or SCID mice that have been reconstituted with human lymphoreticular cells, alters the complexity of cells in the xenografts that serve as potential targets for HIV infection.