Novel Carrier for Polysaccharide Conjugates and an EBV Vaccine

多糖缀合物和 EBV 疫苗的新型载体

• 批准号: 7537173
• 负责人: CLIFFORD M SNAPPER
• 金额: $ 23.09万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-15 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7537173
• 关键词: Adjuvant; Adjuvanticity; Adolescent; Affinity; Anti-Bacterial Agents; Antibodies; Antibody-mediated protection; Antigen Targeting; Antigens; Avidity; B-Lymphocytes; Bacteria; Bacterial Polysaccharides; Binding; Biological Assay; Burkitt Lymphoma; CD4 Positive T Lymphocytes; Carrier Proteins; Clinical Trials; Complement 3d Receptors; Complement Receptor; Conjugate Vaccines; Diphtheria Toxoid; Encapsulated; Enzyme-Linked Immunosorbent Assay; Epstein-Barr Virus Infections; Follicular Dendritic Cells; Goals; Hodgkin Disease; Human; Human Herpesvirus 4; Immunization; Immunocompromised Host; Immunoglobulin G; In Vitro; Incidence; Infant; Infection; Infectious Mononucleosis; Ligands; Link; Macaca mulatta; Measures; Mediating; Morbidity - disease rate; Mus; N-terminal; Nasopharynx Carcinoma; Non-Hodgkin' s Lymphoma; Pathogenesis; Plasma; Polysaccharides; Population; Proteins; Reaction; Receptors, Antigen, B-Cell; Recombinants; Recruitment Activity; Relative (related person); Research Design; Risk; Role; Serotyping; Structure of germinal center of lymph node; Syndrome; T-Lymphocyte; Vaccines; Vertebral column; Viral Proteins; aluminum sulfate; anti-IgG; bactericide; base; cell transformation; env Gene Products; extracellular; immunogenic; in vivo; in vivo Model; meetings; monocyte; mortality; neutralizing antibody; nonhuman primate; novel; preclinical study; prophylactic; response; virus envelope; young adult

DESCRIPTION (provided by applicant): There currently exists a need for novel, immunogenic carrier proteins for new anti-bacterial polysaccharide (PS) conjugate vaccines, as well as a vaccine that is protective against Epstein-Barr virus (EBV) infection. The latter vaccine would have a potential global impact on the incidence of infectious mononucleosis, Hodgkin's and non- Hodgkin's lymphoma, nasopharyngeal carcinoma, and lymphoproliferative syndrome. The EBV protein, gp350, is the major target for EBV neutralizing antibody, as well as a ligand for CD21, a potent co-activator of the B cell antigen receptor. CD21 is also expressed by follicular dendritic cells where it mediates antigen trapping, important for induction of the germinal center reaction. Thus, gp350 in the form of a multimeric gp350-PS conjugate, has the potential to serve as both a potent carrier protein for PS-specific conjugate vaccines as well as an antibody-mediated vaccine for EBV. In preliminary studies we have: 1) expressed and purified a recombinant glycosylated N-terminal 72kDa fragment of the gp350 molecule, 2) conjugated multiple copies of gp350 to pneumococcal capsular polysaccharide, serotype 14 (PPS14) [PPS14-gp350], 3) demonstrated the ability of PPS14-gp350 to specifically bind to CD21 expressed on rhesus and human, but not murine, B cells, and 4) induced boosted plasma IgG anti-PPS14 and IgG anti-gp350 antibodies in young adult rhesus monkeys following i.m. immunization with as little as 0.05 mg of PPS14- gp350 adsorbed on alum. The goal of this proposal is to establish a proof-of-principle in non-human primates, for using gp350 clinically, as a combined novel carrier protein for PS conjugate vaccines and as a protective, antibody-based vaccine for EBV. In this proposal we will utilize young adult rhesus monkeys to: 1) directly compare the ability of alum-adsorbed unconjugated or PPS14-conjugated monomeric or unconjugated dimeric gp350 to elicit high titer and high affinity, EBV-neutralizing anti-gp350 antibody and gp350-specific T cell priming, and 2) directly compare the ability of diphtheria toxoid (DT) [an established carrier protein for conjugate vaccines], conjugated to PPS14, with PPS14- conjugated monomeric gp350 to elicit high titer and high affinity, protective anti-PPS14 antibody, and to determine the role of CD21 binding in the adjuvanticity of gp350 as a carrier protein for PPS14. These pre-clinical studies will form the basis for progressing directly to human clinical trials. Currently, there is no prophylactic vaccine for the Epstein-Barr virus (EBV), which is implicated in the pathogenesis of infectious mononucleosis, nasopharyngeal carcinoma, Burkitt lymphoma, non-Hodgkin's lymphoma, and lymphoproliferative syndrome in immunosuppressed patients. Further, there is also a need for novel protein carriers for polysaccharide conjugate vaccines, which elicit antibody-mediated protection against extracellular bacteria, due to the phenomenon of cross-inhibition. The proposed studies, using the rhesus macaque as an in vivo model, will determine the feasibility of using the EBV envelope protein, gp350, an intrinsically immunostimulatory molecule, as a combined novel carrier for polysaccharide-based conjugate vaccines and as a target antigen for a potent antibody-mediated prophylactic vaccine against EBV infection.

描述（由申请人提供）：目前需要用于新型抗菌多糖（PS）缀合物疫苗的新型免疫原性载体蛋白，以及保护性抗EB病毒（EBV）感染的疫苗。后一种疫苗将对传染性单核细胞增多症、霍奇金和非霍奇金淋巴瘤、鼻咽癌和淋巴组织增生综合征的发病率产生潜在的全球影响。EBV蛋白gp 350是EBV中和抗体的主要靶标，也是CD 21的配体，CD 21是B细胞抗原受体的有效共激活剂。CD 21也由滤泡树突状细胞表达，在那里它介导抗原捕获，这对诱导生发中心反应很重要。因此，多聚体gp 350-PS缀合物形式的gp 350具有充当PS特异性缀合物疫苗的有效载体蛋白以及EBV的抗体介导的疫苗的潜力。在初步研究中，我们有：1）表达并纯化gp 350分子的重组糖基化N-末端72 kDa片段，2）将gp 350的多个拷贝缀合至肺炎球菌荚膜多糖血清型14（PPS 14）[PPS 14-gp 350]，3）证明PPS 14-gp 350特异性结合在恒河猴和人而非鼠上表达的CD 21的能力，B细胞，和4）在年轻成年恒河猴中诱导加强的血浆IgG抗PPS 14和IgG抗gp 350抗体。用吸附在明矾上的少至0.05mg PPS 14-gp 350免疫。本提案的目的是在非人灵长类动物中建立一个原理验证，用于临床使用gp 350作为PS缀合物疫苗的组合新型载体蛋白和作为EBV的保护性抗体疫苗。在本提案中，我们将利用年轻的成年恒河猴：1）直接比较明矾吸附的未缀合的或PPS 14缀合的单体或未缀合的二聚体gp 350引发高滴度和高亲和力、EBV中和的抗gp 350抗体和gp 350特异性T细胞引发的能力，和2）直接比较白喉类毒素（DT）[一种用于缀合疫苗的确定载体蛋白]的能力，与PPS 14缀合的单体gp 350，以引发高滴度和高亲和力的保护性抗PPS 14抗体，并确定CD 21结合在gp 350作为PPS 14载体蛋白的佐剂性中的作用。这些临床前研究将成为直接进入人体临床试验的基础。目前，没有针对EB病毒（EBV）的预防性疫苗，EB病毒与免疫抑制患者中的传染性单核细胞增多症、鼻咽癌、伯基特淋巴瘤、非霍奇金淋巴瘤和淋巴组织增生综合征的发病机制有关。此外，还需要用于多糖缀合物疫苗的新型蛋白质载体，由于交叉抑制现象，其引发抗体介导的针对细胞外细菌的保护。所提出的研究，使用恒河猴作为体内模型，将确定使用EBV包膜蛋白，gp 350，一种内在的免疫刺激分子，作为一种组合的新型载体的多糖为基础的缀合物疫苗和作为一种有效的抗体介导的预防性疫苗抗EBV感染的靶抗原的可行性。

项目成果

A novel tetrameric gp350 1-470 as a potential Epstein-Barr virus vaccine.

• DOI: 10.1016/j.vaccine.2013.04.071
• 发表时间: 2013-06-26
• 期刊: VACCINE
• 影响因子: 5.5
• 作者: Cui, Xinle;Cao, Zhouhong;Sen, Goutam;Chattopadhyay, Gouri;Fuller, Deborah H.;Fuller, James T.;Snapper, Dustin M.;Snow, Andrew L.;Mond, James J.;Snapper, Clifford M.
• 通讯作者: Snapper, Clifford M.