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.

50亿人患有牙科疾病。基于广泛的公共卫生反卡里措施是必要的 在征服这种疾病之前。龋齿，尤其是在童年时期，与水平密切相关 突变群链球菌定殖，尤其是S. utans和S. sobrinus。尽管进行了深入调查，但 关于突变链球菌的初始定植和积累是否存在 一二岁的孩子的口服生物膜可能会受到抗原抗原抗体的影响 诱变链球菌，例如通过“被动”免疫疗法。这种延续的目的 04733将开发3个高亲和力人类抗葡萄糖基转移酶（GIF）单克隆试剂 小儿免疫治疗用途。为此，我们将合并我们在识别保护性表位的经验 GTF以及我们成功应用主动和被动免疫方法来减少实验 龋齿，具有Marasco/Dana Farber实验室成功发展人类的能力 人类疾病的单克隆试剂（HMR）。为此，我们建议发展高亲和力，抑制性 HMR通过与经过验证的270亿成员人类的隔离为针对S. Mutans GTF表位 单链变量区域抗体片段（SCFV）噬菌体显示库，随后的二价SCFVFC 抗体（小动物）结构，表位图和亲和力确定以最大化抑制作用 潜力，然后是全人类单克隆抗体（HMR）结构。抑制GTF酶促的 人工生物膜中链球菌链球菌的积累的活性和干扰将用于 测量SCFV噬菌体，SCFVFC在HMR开发过程中对GTF功能活性的体外影响。我们 然后，将通过使用这些HMR来评估体内免疫治疗候选物的保护潜力 单独和集体，以阻止两者的殖民化，积累和龋齿形成能力 感染人类的​​链球菌种类。该协议将在3个目标下执行：隔离一个 GTF抑制性HMR面板；确定HMR阻止突变链球菌进入进入的能力 生物膜并确定HMR干扰大鼠龋齿的能力。一旦开发和审查 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小儿免疫治疗 阻止或延迟链球菌的口腔定植/积累的方法。