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

• 批准号: 10737872
• 负责人: Javier Gordon Ogembo
• 金额: $ 7.4万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 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/10737872
• 关键词: 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 Cells; Epithelium; Fibroblasts; Future; Glycoproteins; Goals; HIV/AIDS; 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; Predisposition; 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; cell type; design; experimental study; human model; immunosuppressed; in vivo; in vivo Model; latent infection; mutant; nonhuman primate; novel; oral infection; permissiveness; polyclonal antibody; polyclonal human antibody; prevent; prophylactic; receptor; skin lesion; success; tool; transmission process; vaccine candidate; vaccine development

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.

项目摘要 每年全球报告了44,000多个Kaposi肉瘤（KS），其中84％发生在 非洲。这和其他Kaposi肉瘤相关的疱疹病毒（KSHV）引起的恶性肿瘤占主导 尽管可以在其他人体中检测到KSHV 流体，其经常在唾液中以有或没有性传播感染的风险（例如， 儿童）建议口腔是初次收购的地点。但是，KSHV口服的机制 体内传播，尤其是病毒进入所需的关键病毒包膜糖蛋白（GPS）， 仍然无法解决。已经确定了几次KSHV - HOST相互作用，但所有先前的实验均为 由于缺乏适当的动物模型，因此在体外进行的，尚未在体内验证。通过 与威斯康星州国家灵长类动物研究中心合作，我们的实验室可以使用共同 Marmoset（Callithrix Jacchus，CJ），最近开发的KSHV非人类私人模型，容易受到影响 KSHV口腔感染，在免疫抑制下会获得KS样皮肤病变。这个目的 应用是阐明在体内启动原发性口腔感染所需的最低GPS，作为前提条件 选择用于开发有效预防性疫苗候选者的关键GP。该应用程序建立在Dr. Ogembo最近完成了NCI K01 CA184388-05研究KSHV进入机制和疫苗的研究 发展。最近，我们表明在体外，KSHV糖蛋白GH/GL对于病毒感染至关重要 上皮，内皮和成纤维细胞，而不是B细胞。值得注意的是，我们和其他人也表明 单克隆和多克隆ABS至KSHV糖蛋白GB，GH/GL和GPK8.1，可以中和KSHV感染 潜水员在体外允许的人类细胞。在成功的基础上，我们产生了缺乏的KSHV删除突变体 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 特定于这些GP。在这个项目中，我们将使用人体离体样本和CJ KSHV模型来测试 假设GB和GH/GL对于KSHV体内口服至关重要。我们建议的前提是 建立在有力的证据的基础上，表明1）KSHV可以感染CJ，会产生类似KS的皮肤病变，以及2）ABS针对 KSHV糖蛋白GB和GH/GL可以在体外和体内中和KSHV感染。此外， 人体细胞的KSHV感染的允许性，使这些平台非常适合测试KSHV GP感染要求。成功完成拟议的研究将阐明最小KSHV 在离体和体内模型中产生原发性感染所需的GPS，促进了我们定义的长期目标 人类KSHV感染的初始步骤以及抗体在保护KSHV早期步骤中的作用 传播。这最终将为预防性疫苗的设计和开发提供信息，以防止KSHV 感染及其相关的癌症。