Passive Immunotherapy for Pediatric Mutans Streptococcal Infections

小儿变形链球菌感染的被动免疫治疗

• 批准号: 7464022
• 负责人: MARTIN A TAUBMAN
• 金额: $ 108.16万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7464022
• 关键词: 2 year old; Affinity; Antibodies; Antigens; Bacteriophages; Binding; Child; Childhood; Condition; Dental caries; Development; Disease; Dose; Epitope Mapping; Epitopes; Experimental Animal Model; Facility Construction Funding Category; Glucosyltransferase; Glucosyltransferases; Goals; Human; Immune; Immunoglobulin Fragments; Immunoglobulin Variable Region; Immunotherapeutic agent; In Vitro; Infection; Investigation; Laboratories; Libraries; Measures; Microbial Biofilms; Modeling; Monoclonal Antibodies; Oral; Passive Immunization; Passive Immunotherapy; Phage Display; Protocols documentation; Public Health; Rattus; Reagent; Streptococcal Infections; Streptococcus mutans; Tooth Diseases; Virulence; base; experience; human disease; human monoclonal antibodies; in vivo; man; member; oral biofilm; response; tool

Five billion people suffer from dental disease. Broad-based public health anti-caries measures are necessary before this disease is conquered. Dental caries, especially in childhood, is strongly correlated with levels of mutans streptococcal colonization, particularly S. mutans and S. sobrinus. Despite intensive investigation, a major gap exists regarding whether the initial colonization and accumulation of mutans streptococci in the oral biofilm of the one-two year old child can be influenced by the presence of antibody to virulence antigens of mutans streptococci, for example through "passive" immunotherapy. The goal of this continuation of DE- 04733 is to develop up to 3 high affinity human anti-glucosyltransferase (GIF) monoclonal reagents for pediatric immunotherapeutic use. To do so we will merge our experience in identifying protective epitopes on GTF and our successful application of active and passive immune approaches to reduce experimental dental caries, with the proven ability of the Marasco/Dana Farber laboratories to successfully develop human monoclonal reagents (HMRs) for human diseases. To this end we propose to develop high-affinity, inhibitory HMRs directed against S. mutans GTF epitopes through isolation from a validated 27 billion member human single-chain variable region antibody fragment (scFv) phage display library, subsequent bivalent scFvFc antibody (minibody) construction, epitope mapping and affinity determination to maximize inhibition potential, followed by full human monoclonal antibody (HMR) construction. Inhibition of GTF enzymatic activity and interference with accumulation of mutans streptococci in artificial biofilms will be used to measure in vitro effects on GTF functional activity by scFv phage, scFvFc during HMR development. We will then evaluate the protective potential of the immunotherapeutic candidates in vivo, by using these HMRs, individually and collectively, to block colonization, accumulation and dental caries-forming ability of both mutans streptococcal species that infect man. This protocol will be performed under 3 aims: To isolate a panel of GTF-inhibitory HMR; to determine the ability of HMR to block mutans streptococcal entry into biofilm and to determine the ability of HMR to interfere with dental caries in rats. Once developed and vetted through our experimental animal model, we will be poised to use these tools to pursue pediatric immunotherapeutic approaches to block or delay the oral colonization/accumulation of cariogenic mutans streptococci.

五十亿人患有牙科疾病。基础广泛的公共卫生防龋措施是必要的 在这种疾病被征服之前。龋齿，尤其是儿童时期的龋齿，与龋齿水平密切相关。 变形链球菌定植，特别是变形链球菌和远缘链球菌。尽管进行了深入调查， 关于变形链球菌是否在体内最初定植和积累，存在重大分歧。 一两岁儿童的口腔生物膜可能会受到毒力抗原抗体的影响 变形链球菌，例如通过“被动”免疫疗法。 DE 延续的目标是 04733 开发多达3种高亲和力人抗葡萄糖基转移酶（GIF）单克隆试剂 儿科免疫治疗用途。为此，我们将结合我们在识别保护性表位方面的经验 GTF 和我们成功应用主动和被动免疫方法来减少实验 龋齿，Marasco/Dana Farber 实验室已证明有能力成功开发人类 用于人类疾病的单克隆试剂（HMR）。为此，我们建议开发高亲和力、抑制性的 通过从经过验证的 270 亿人类成员中分离出针对变形链球菌 GTF 表位的 HMR 单链可变区抗体片段 (scFv) 噬菌体展示文库，后续二价 scFvFc 抗体（微型抗体）构建、表位作图和亲和力测定，以最大限度地抑制 潜力，然后是完整的人单克隆抗体（HMR）构建。 GTF酶的抑制 人工生物膜中变形链球菌的活性和对积累的干扰将用于 测量 HMR 发育过程中 scFv 噬菌体、scFvFc 对 GTF 功能活性的体外影响。我们 然后将通过使用这些 HMR 评估免疫治疗候选药物的体内保护潜力， 单独和集体地阻止两者的定殖、积累和龋齿形成能力 感染人类的​​变形链球菌。该协议将在 3 个目标下执行： GTF 抑制性 HMR 组；确定 HMR 阻止变形链球菌进入体内的能力 生物膜并确定 HMR 干扰大鼠龋齿的能力。一旦开发并经过审查 通过我们的实验动物模型，我们将准备使用这些工具来追求儿科免疫治疗 阻止或延迟致龋变形链球菌口腔定植/积累的方法。