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：
10623087
负责人：
Javier Gordon Ogembo
金额：
$ 12.5万
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/10623087
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-22-036. 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.

项目摘要此申请是根据特殊利益通知（NOSI）提交的被确定为非CA-22-036。据报道，全球有超过44,000例卡波西肉瘤（KS）一年，其中84％发生在非洲。这和其他Kaposi肉瘤相关的疱疹病毒（KSHV）引起的虽然KSHV可以是在其他人体液体中检测到，其经常以有或没有风险的人群中的唾液检测性传播感染（例如，儿童）表明口腔是主要收购的部位。但是，体内KSHV口服传播的机制，尤其是关键病毒膜病毒进入所需的糖蛋白（GPS）仍未解决。几个KSHV – HOST互动具有被鉴定出来，但是所有先前的实验均在体外进行，并且由于先验而在体内尚未得到验证缺乏合适的动物模型。通过与威斯康星州国家灵长类动物研究的合作中心，我们的实验室可以使用最近开发的KSHV的Common Marmoset（Callithrix Jacchus，CJ）易受KSHV口腔感染的非人类私人模型，在免疫抑制下获得了类似KS的皮肤病变。该应用的目的是阐明启动初级所需的最低GP 体内口服感染，作为选择用于开发有效预防性疫苗的关键GP的先决条件候选人。该申请基于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，我们表明在体外，KSHV糖蛋白 GH/GL对于上皮，内皮细胞和成纤维细胞的病毒感染至关重要，而不是B细胞。值得注意的是，我们其他人也表明，单克隆和多克隆ABS均与KSHV糖蛋白GB，GH/GL和 GPK8.1可以在体外中和潜水员允许的人类细胞的KSHV感染。在这一成功的基础上，我们缺少四种糖蛋白的KSHV缺失突变体对病毒进入至关重要（GB，GH/GL，， GPK8.1）和对这些GPS特异的各种单克隆抗体。在这个项目中，我们将使用人类的体内样品和CJ KSHV模型，以测试GB和GH/GL对于体内KSHV至关重要的假设传播。我们提案的前提是基于有力证据，表明1）KSHV可以感染CJ，而CJ KS样皮肤病变和2）ABS针对KSHV糖蛋白GB和GH/GL可以中和KSHV感染体外和体内。此外，人类细胞的KSHV感染允许在体内和CJ中使这些允许平台非常适合测试KSHV GP感染要求。拟议的研究成功完成将阐明离体和体内模型中原发性感染所需的最小KSHV GP 定义人类KSHV感染的初始步骤以及抗体在保护中的作用的长期目标违反KSHV传输的早期步骤。这最终将为预防性的设计和开发提供信息可以防止KSHV感染及其相关癌症的疫苗。