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）或较不常见的单纯疱疹病毒1型（HSV-1）引起的。该提案将侧重于HSV-2。女性的感染可以针对阴唇表面、阴道和子宫颈。皮肤的邻近区域也可能被感染。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期，并可作为复发感染的来源。原发性感染通常更严重，但复发性感染可能发生数年，并作为一个潜在的传染性病毒来源。这种病毒感染的发病率非常高。免疫抑制的患者在所有组织中都有这种病毒复制的严重风险，并可能遭受危及生命的后遗症。目前治疗HSV- 2感染的主要药物是阿昔洛韦，一种有效的核苷酸类似物，特异性磷酸化并掺入HSV感染细胞的DNA中。该化合物的静脉、口服和局部制剂具有治疗益处。此外，某些基于去污剂的杀精子剂已被证明对HSV具有抗病毒活性。本计划项目前三年的研究表明，C31 G（C14/C16）和烷基硫酸盐杀微生物剂均可杀灭HSV-2。重要的是，SDS已显示在小鼠阴道感染的体内模型中预防HSV-2感染，SDS和C31 G已显示在人阴道异种移植模型中抑制HSV-2并预防感染。我们在下一阶段的具体目标将包括：1）继续采用三种HSV-2生长模型系统，以确定非配制和配制的杀微生物化合物的毒性和功效：a）通过猴肾上皮细胞和原代人阴道角质形成细胞中的噬斑形成来体外测定HSV-2; B）通过阴道接种Swiss-Webster远系杂交小鼠进行体内测定，c）通过接种在免疫受损小鼠中生长的人阴道异种移植物进行体内测定; 2）比较接种正常人阴道异种移植物或表达来自HPV-11的病毒基因的完整库的人阴道异种移植物后HSV-2感染的动力学和自然史;和3）确定在裸鼠、SCID小鼠或用人淋巴网状细胞重建的SCID小鼠中生长的人阴道异种移植物的急性或亚临床HSV-2感染是否改变异种移植物中作为HIV感染的潜在靶的细胞的复杂性。

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.