Oral and Vaginal Gene Expression by Candida During AIDS

描述（由申请人提供）：口咽和外阴阴道念珠菌病（OPC和VVC），由白色念珠菌引起的粘膜感染，是HIV感染者中主要的机会性感染。在大多数人中，C.白色念珠菌是胃肠道的无害寄生物。生物体转变为入侵性病原体的机制知之甚少。我们推测，在口腔和阴道粘膜的发病机制是促进了一个子集的念珠菌基因的表达是特定的粘膜表面。我们实验室的主要总体目标是了解C。人类感染致病过程中白念珠菌基因的表达。这个R21项目的目标是识别C。在口腔和/或阴道粘膜特异性表达的白色念珠菌基因，以鉴定有助于HIV相关念珠菌病发病的基因。为了实现这一点，我们将使用唾液和阴道液，富含当地产生的抗念珠菌抗体，以确定免疫原性C。白色念珠菌抗原在初步研究中，我们分别从活动性OPC和VVC的HIV感染者中创建了单独的唾液和阴道液池。混合样品对酵母菌和菌丝的吸附。体外生长的白色念珠菌菌株以及细胞裂解物。在特异性目标1中，我们将使用吸附的样品，现在集中在针对人类宿主中独特表达的抗原的抗体中，以筛选C。白色念珠菌基因组表达文库。编码免疫原性抗原的基因的完整序列将使用C.白色念珠菌序列数据库。在特定目标2中，我们将使用定量实时逆转录-聚合酶链反应证实这些基因在口腔或阴道粘膜诱导，但在体外常规生长期间不诱导。我们还将通过使用针对纯化抗原产生的常规抗体来探测从HIV感染者中回收的OPC和VVC样本，直接检测体内诱导的抗原。本项目中鉴定的粘膜特异性基因将在未来的项目中研究其在念珠菌发病机制中的特定作用，最终目标是设计治疗和预防OPC和VVC的策略。

DESCRIPTION (provided by applicant): Oropharyngeal and vulvovaginal candidiasis (OPC and VVC), mucosal infections caused by Candida albicans, are the leading opportunistic infections among HIV-infected persons. In most humans, C. albicans is a harmless commensal of the gastrointestinal tract. The mechanisms by which the organism switches to an invasive pathogen are poorly understood. We hypothesize that pathogenesis at the oral and vaginal mucosa is facilitated by the expression of a subset of candidal genes that are specific to the mucosal surfaces. The major overall objective of our laboratory is to understand C. albicans gene expression during the pathogenesis of human infections. The goal of this R21 project is to identify C. albicans genes that are specifically expressed at the oral and/or vaginal mucosa, in order to identify genes that contribute to the pathogenesis of HIV associated candidiasis. To accomplish this, we will use saliva and vaginal fluid that are rich in locally produced anti-candidal antibodies to identify immunogenic C. albicans antigens. In preliminary studies, we created separate pools of saliva and vaginal fluid from HIV-infected persons with active OPC and VVC, respectively. The pooled samples were adsorbed against yeast and hyphae of C. albicans strains grown in vitro, as well as cell lysates. In Specific Aim 1, we will use the adsorbed samples, now concentrated in antibodies directed against antigens expressed uniquely within the human host, to screen a C. albicans genomic expression library in parallel. The complete sequences of the genes encoding immunogenic antigens will be identified using the C. albicans sequence databases. In Specific Aim 2, we will confirm that these genes are induced at the oral or vaginal mucosa, but not during routine growth in vitro, using quantitative real-time reverse transcription-polymerase chain reaction. We will also directly detect the in vivo induced antigens by using routine antibodies raised against the purified antigens to probe OPC and VVC samples recovered from HIV-infected persons. The mucosal-specific genes identified in this project will be studied in future projects for their specific roles during candidal pathogenesis, with the ultimate goal of designing strategies to treat and prevent OPC and VVC.