Unmasking the roles of viral glycoproteins in oral transmission of KSHV

揭示病毒糖蛋白在 KSHV 口腔传播中的作用

基本信息

批准号：
10318876
负责人：
Javier Gordon Ogembo
金额：
$ 85.4万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10318876
关键词：
AIDS/HIV problem Africa Age Animal Model Antibodies Antibody-mediated protection B-Lymphocytes Binding Body Fluids Callithrix jacchus jacchus Cell surface Cells Child Collaborations Complex Data Detection Development Disease Endothelial Cells Endothelium Epithelial Epithelial Cells Fibroblasts Future Glycoproteins Goals Herpesviridae Herpesviridae Infections Human Human Herpesvirus 8 Human body Iatrogenesis Immunity Immunize Immunocompetent Immunocompromised Host Immunologic Deficiency Syndromes Immunosuppression In Vitro Infection Kaposi Sarcoma Laboratories Lesion Lymphocyte Malignant Neoplasms Malignant lymphoid neoplasm Maternal antibody Measures Membrane Modeling Monoclonal Antibodies Oral Oral cavity Pattern Preventive vaccine Primary Infection Primates Receptor Cell Recombinants Reporting Research Risk Role Saliva Sampling Sexually Transmitted Diseases Signal Transduction Site T-Lymphocyte Testing Tonsil Transplant Recipients Tropism Viral Viremia Virion Virus Diseases Wisconsin Work antibody test base cell type design experimental study human model immunosuppressed in vivo in vivo Model latent infection mutant nonhuman primate novel oral infection permissiveness polyclonal antibody prevent prophylactic receptor skin lesion success tool transmission process vaccine candidate vaccine development

项目摘要

PROJECT SUMMARY More than 44,000 new cases of Kaposi sarcoma (KS) are reported globally each year, 84% of which occur in Africa. This and other Kaposi sarcoma-associated herpesvirus (KSHV)-induced malignancies predominate in people with acquired or iatrogenic immunodeficiencies. Although KSHV can be detected in other human body fluids, its frequent detection in saliva in groups both with and without risk of sexually transmitted infections (e.g., children) suggests that the oral cavity is the site of primary acquisition. However, the mechanism of KSHV oral transmission in vivo, particularly the critical viral envelope glycoproteins (gps) required for viral entry, remains unresolved. Several KSHV–host interactions have been identified, but all prior experiments were performed in vitro and have not been validated in vivo due to prior lack of an appropriate animal model. Through collaboration with the Wisconsin National Primate Research Center, our laboratory has access to the common marmoset (Callithrix jacchus, CJ), a recently developed KSHV non-human primate model that is susceptible to KSHV oral infection, and under immunosuppression acquires KS-like skin lesions. The objective of this application is to elucidate the minimum gps required to initiate primary oral infection in vivo, as a prerequisite to selecting key gps for developing an effective prophylactic vaccine candidate. This application builds on Dr. Ogembo’s recently completed NCI K01 CA184388-05 research on KSHV entry mechanisms and vaccine development. Recently, we showed that in vitro, the KSHV glycoprotein gH/gL is essential for viral infection of epithelial, endothelial, and fibroblasts cells, but not B cells. Notably, we and others have also shown that both monoclonal and polyclonal Abs to KSHV glycoproteins gB, gH/gL, and gpK8.1, can neutralize KSHV infection of diverse permissive human cells in vitro. Building on this success, we generated KSHV deletion mutants lacking the four glycoproteins thought to be critical for viral entry (gB, gH/gL, gpK8.1) and various monoclonal antibodies specific to these gps. In this project, we will use human ex vivo samples and the CJ KSHV model to test the hypothesis that gB and gH/gL are critical for KSHV in vivo oral transmission. The premise of our proposal is built on strong evidence that 1) KSHV can infect CJ, which develop KS-like skin lesions, and 2) Abs against the KSHV glycoproteins gB and gH/gL can neutralize KSHV infection in vitro and ex vivo. Furthermore, the permissiveness to KSHV infection of human cells ex vivo and CJ makes these platforms ideal to test the KSHV gp requirements for infection. Successful completion of the proposed study will elucidate the minimum KSHV gps required for primary infection in ex vivo and in vivo models, advancing our long-term goal of defining the initial steps in KSHV infection of humans and the role of antibodies in protecting against the early steps of KSHV transmission. This will ultimately inform design and development of prophylactic vaccines that can prevent KSHV infection and its associated cancers.

项目概要全球每年报告超过 44,000 例卡波西肉瘤 (KS) 新病例，其中 84% 发生在这种疾病和其他卡波西肉瘤相关疱疹病毒 (KSHV) 引起的恶性肿瘤在非洲占主导地位。尽管 KSHV 可以在其他人体中检测到，但患有获得性或医源性免疫缺陷的人。在有或没有性传播感染风险的群体（例如，儿童）表明口腔是主要感染部位，但 KSHV 口腔感染的机制尚不清楚。体内传播，特别是病毒进入所需的关键病毒包膜糖蛋白（gps），已经确定了一些 KSHV 与宿主的相互作用，但所有先前的实验均已确定。由于之前缺乏合适的动物模型，该研究在体外进行，尚未在体内得到验证。我们的实验室与威斯康星州国家灵长类动物研究中心合作，可以获得共同的狨猴（Callithrix jacchus，CJ），一种最近开发的 KSHV 非人类灵长类动物模型，易受 KSHV 口腔感染，并在免疫抑制下获得 KS 样皮肤损害。应用是为了阐明在体内引发原发性口腔感染所需的最低 GPS，作为先决条件选择关键的 GPS 来开发有效的预防性候选疫苗。 Ogembo最近完成了关于KSHV进入机制和疫苗的NCI K01 CA184388-05研究最近，我们在体外证明，KSHV 糖蛋白 gH/gL 对于病毒感染至关重要。上皮细胞、内皮细胞和成纤维细胞，但不包括 B 细胞。值得注意的是，我们和其他人还表明，两者都存在。针对 KSHV 糖蛋白 gB、gH/gL 和 gpK8.1 的单克隆和多克隆抗体，可以中和 KSHV 感染在这一成功的基础上，我们在体外产生了多种许可的人类细胞，从而产生了缺乏 KSHV 的突变体。被认为对病毒进入至关重要的四种糖蛋白（gB、gH/gL、gpK8.1）和各种单克隆抗体针对这些 GPS，我们将使用人体离体样本和 CJ KSHV 模型来测试。 gB 和 gH/gL 对于 KSHV 体内口腔传播至关重要的假设我们提出的前提是。建立在强有力的证据之上，即 1) KSHV 可以感染 CJ，从而出现 KS 样皮肤损伤，以及 2) 针对 KSHV 的抗体 KSHV 糖蛋白 gB 和 gH/gL 可以在体外和离体中和 KSHV 感染。允许 KSHV 体外感染人体细胞和 CJ，使这些平台成为测试 KSHV 的理想选择成功完成拟议的研究将阐明最低 KSHV。离体和体内模型中原发感染所需的 GPS，推进了我们定义感染的长期目标人类 KSHV 感染的初始步骤以及抗体在预防 KSHV 早期步骤中的作用这最终将为预防 KSHV 的预防性疫苗的设计和开发提供信息。感染及其相关癌症。