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：
10627167
负责人：
Javier Gordon Ogembo
金额：
$ 20万
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/10627167
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 Persons 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 base cell type design experimental study human model immunosuppressed in vivo in vivo Model interest latent infection mutant nonhuman primate novel oral infection permissiveness polyclonal antibody prevent prophylactic receptor response skin lesion success tool transmission process vaccine candidate vaccine development

项目摘要

PROJECT SUMMARY This application is being submitted in response to the Notice of Special Interest (NOSI) identified as “NOT-CA-22-057.” 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.

项目摘要本申请是为了响应特别利益通知 (NOSI) 而提交的被识别为“NOT-CA-22-057”。全球每年报告超过 44,000 例卡波西肉瘤 (KS) 新病例今年，其中 84% 发生在非洲，以及其他卡波西肉瘤相关疱疹病毒 (KSHV) 引起的。尽管 KSHV 也可能是获得性或医源性免疫缺陷患者的恶性肿瘤。在其他人体体液中检测到，在有或没有风险的群体的唾液中经常检测到它性传播感染（例如儿童）表明口腔是主要感染部位。然而，KSHV在体内经口传播的机制，特别是关键的病毒包膜病毒进入所需的糖蛋白 (gps) 的一些问题仍未得到解决。已被确定，但所有先前的实验都是在体外进行的，并且由于先前的原因尚未在体内得到验证通过与威斯康星国家灵长类动物研究中心的合作，缺乏合适的动物模型。中心，我们的实验室可以使用普通狨猴（Callithrix jacchus，CJ），这是一种最近开发的 KSHV 易受 KSHV 口腔感染且在免疫抑制下获得的非人灵长类动物模型 KS 样皮肤病变的目的是阐明启动原发性所需的最低 GPS 值。体内口腔感染，作为选择关键 gps 开发有效预防性疫苗的先决条件该应用程序基于 Ogembo 博士最近完成的 NCI K01 CA184388-05 研究。 KSHV 进入机制和疫苗开发最近，我们在体外证明了 KSHV 糖蛋白。 gH/gL 对于上皮细胞、内皮细胞和成纤维细胞的病毒感染至关重要，但对于 B 细胞则不然。等人还表明，针对 KSHV 糖蛋白 gB、gH/gL 和 gpK8.1，可以在体外中和 KSHV 感染多种允许的人类细胞，在此基础上，我们。生成的 KSHV 缺失突变体缺乏被认为对病毒进入至关重要的四种糖蛋白（gB、gH/gL、 gpK8.1）和针对这些 gps 的各种单克隆抗体在该项目中，我们将使用人类离体。样本和 CJ KSHV 模型来检验 gB 和 gH/gL 对 KSHV 体内口服至关重要的假设我们建议的前提是建立在强有力的证据之上：1) KSHV 可以感染 CJ，而 CJ 会发展。 KS 样皮肤损伤，2) 针对 KSHV 糖蛋白 gB 和 gH/gL 的抗体可以中和 KSHV 感染此外，体外和离体人体细胞对 KSHV 感染的允许性使这些成为可能。测试 KSHV gp 感染要求的理想平台将成功完成拟议的研究。阐明离体和体内模型中原发感染所需的最低 KSHV gps，推进我们的研究确定人类 KSHV 感染的初始步骤以及抗体在保护中的作用的长期目标针对 KSHV 传播的早期步骤，这最终将为预防性药物的设计和开发提供信息。可以预防 KSHV 感染及其相关癌症的疫苗。