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: 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
• 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

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 本申请是为了响应特别利益通知 (NOSI) 而提交的 标识为 NOT-CA-22-036。全球每年报告超过 44,000 例卡波西肉瘤 (KS) 新病例 年，其中 84% 发生在非洲。该病毒和其他卡波西肉瘤相关疱疹病毒 (KSHV) 诱导的 恶性肿瘤在患有获得性或医源性免疫缺陷的人群中占主导地位。虽然 KSHV 可以 在其他人体体液中检测到，无论有或没有风险的群体的唾液中都经常检测到它 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）仍然悬而未决。一些 KSHV 与宿主的相互作用 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 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中心合作 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样皮损。此应用程序的目的是阐明启动主要功能所需的最低 GPS 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 候选人。该应用程序建立在 Ogembo 博士最近完成的 NCI K01 CA184388-05 研究的基础上 KSHV 进入机制和疫苗开发。最近，我们在体外证明，KSHV 糖蛋白 gH/gL 对于上皮细胞、内皮细胞和成纤维细胞（但 B 细胞）的病毒感染至关重要。值得注意的是，我们 等人还表明，针对 KSHV 糖蛋白 gB、gH/gL 和 gpK8.1，可以在体外中和多种允许的人类细胞的 KSHV 感染。在此成功的基础上，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。在这个项目中，我们将使用人类离体 样本和 CJ KSHV 模型来检验 gB 和 gH/gL 对于 KSHV 体内口服至关重要的假设 传播。我们提议的前提是建立在强有力的证据之上，即 1) KSHV 可以感染 CJ，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 体外和离体。此外，人体细胞离体和 CJ 允许 KSHV 感染，使得这些 测试 KSHV gp 感染要求的理想平台。成功完成拟议的研究将 阐明体外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 反对 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。