Scanning the Cryptosporidium proteome for vaccine antigens

扫描隐孢子虫蛋白质组寻找疫苗抗原

• 批准号: 10011112
• 负责人: Joseph J Campo
• 金额: $ 30万
• 依托单位: IMMPORT THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011112
• 关键词: AIDS/HIV problem; Address; Antibodies; Antibody Response; Antigens; Applications Grants; Baculoviruses; Bangladeshi; Binding; Biological Assay; Birth; Breast Cancer Detection; Breastfed infant; Budgets; California; Cessation of life; Child; Childhood; Clinical; Collaborations; Communities; Core Protein; Cryptosporidiosis; Cryptosporidium; Detection; Development; Diarrhea; Disease; Enzyme-Linked Immunosorbent Assay; Eukaryotic Cell; Foundations; Future; Human Milk; Immune; Immune response; Immunization; Immunocompromised Host; Immunoglobulin A; Incidence; Infant; Infantile Diarrhea; Infection; Ingestion; Intervention; Knowledge; Life; Link; Malnutrition; Manufacturer Name; Measures; Morbidity - disease rate; Mothers; Mucosal Immune Responses; Mucous Membrane; Neurocognitive Deficit; Oocysts; Open Reading Frames; Oral; Parasites; Phase; Post-Translational Protein Processing; Postpartum Period; Prevention; Protein Array; Protein Microchips; Proteins; Proteome; Proteomics; Research; Risk; Sampling; Scanning; Small Business Innovation Research Grant; Spottings; Structure; Supplementation; System; Technology; Testing; Therapeutic antibodies; TimeLine; Universities; Vaccine Antigen; Vaccines; Validation; Virginia; Western Blotting; Work; adaptive immunity; chronic infection; clinical development; cohort; commercialization; disability-adjusted life years; efficacy study; follow-up; high risk infant; immunogenicity; immunoreactivity; infancy; infection burden; low and middle-income countries; mortality; mouse model; pathogen; pre-clinical; preclinical study; prevent; programs; response; success; technology validation; tool; vaccination schedule; vaccine candidate; vaccine development; vaccinology; years of life lost

Cryptosporidiosis is a top ten cause of infant diarrhea in low and middle-income countries (LMICs), as well as malnutrition and impaired neurocognitive development, leading to substantial yearly morbidity, mortality and lost disability-adjusted life years. A vaccine for cryptosporidiosis is not available, but would be appealing for prevention of cryptosporidiosis in children in LMICs where the burden of infection is high. Studies done by Dr. William A. Petri and Dr. Carol Gilchrist at University of Virginia have uncovered IgA correlates of protection against Cryptosporidium infection, which may be expoited for development of a vaccine. However, the cryptosporidial proteins targeted by protective IgA antibodies remain to be discovered. Antigen Discovery, Inc (ADI) of Irvine, California has developed a pilot scale Cryptosporidium protein microarray, which can be expanded and used to screen antibody responses against the cryptosporidial proteome. A proteome-scale platform for antibody immune-profiling has never before been available to the cryptosporidiosis research community, and this technology has the power to rapidly advance our understanding of the protective immune response directed to cryptosporidial proteins. A pan-proteome Cryptosporidium microarray will be developed to measure specific anti-cryptosporidial IgA levels in two mother-infant birth cohorts with years of follow-up and detailed characterization of clinical endpoints for diarrhea, including disease attributable detection of many diarrhea-causing pathogens, including Cryptosporidium spp. The aim of the study is to identify the IgA in maternal breast milk ingested by infants that correlates with reduced risk of subsequent Cryptosporidium infection and diarrhea. The array results and most promising vaccine candidates will be validated by producing the proteins in a eukaryotic expression system to include post-translational modifications and correct tertiary structure, which will be used to develop ELISA and Western blot assays for testing breast milk samples. The success of this study will present an opportunity for development of maternal vaccines to prevent cryptosporidiosis in infants during the vulnerable period of the first months of life. We postulate that protective antibodies identified in breast milk may also be protective when produced by the infant mucosal immune response, thus presenting the opportunity for development of a pediatric vaccine for continued protection. Protective antibodies may also be developed as therapeutic antibodies that can be given to treat persistent infection in immunocompromised patients, such as those with HIV/AIDS. We expect to identify 240-400 immunoreactive Cryptosporidium proteins, at least 3 of which will have increased IgA levels in the breast milk of mothers with infants that had lower incidence rates or protection from diarrhea, as well as fecal IgA from the infants’ adaptive immunity. This grant application addresses the significant problem of cryptosporidiosis in children by laying the foundation for a vaccine through the study of specific mucosal IgA responses associated with protection.

隐孢子虫病是低收入和中等收入国家（LMIC）的婴儿腹泻的十大原因，也是 营养不良和神经认知发展受损，导致年度大量发病率，死亡率和丧失 残疾调整后的生活年。无法使用隐孢子虫病的疫苗，但会出现 预防感染燃烧的LMIC儿童中隐孢子虫病。 Dr. 弗吉尼亚大学的William A. Petri和Carol Gilchrist博士发现了IGA保护的相关性 反对隐孢子虫感染，可能暴露于疫苗的开发。但是， 受保护的IgA抗体靶向的隐孢子型蛋白仍有待发现。 Antigen Discovery，Inc 加利福尼亚州欧文的（ADI）开发了一个试验量表隐孢子虫蛋白微阵列，可以是 扩展并用于筛选针对隐孢子虫蛋白质组的抗体反应。蛋白质组规模 抗体免疫促进平台从来没有用于隐孢子虫病研究 社区，这项技术有能力快速提高我们对受保护免疫的理解 反应针对隐孢子虫蛋白。将开发一个泛蛋白酶隐孢子虫微阵列 测量两个母亲的抗孢子体IgA水平，包括多年的随访和 腹泻的临床终点的详细表征，包括疾病可归因于许多 引起腹泻的病原体，包括隐孢子虫属。该研究的目的是识别母体中的IgA 母乳是由婴儿摄入的，与随后的隐孢子虫感染的风险降低相关 腹泻。阵列结果和最有前途的疫苗候选物将通过在 一个真核表达系统，包括翻译后修饰和正确的三级结构，该系统 将用于开发ELISA和Western印迹测定法以测试母乳样品。这个成功 研究将为开发孕产妇疫苗的开发机会预防婴儿隐孢子虫病 在生命的头几个月的脆弱时期。我们假设在乳房中鉴定出的受保护抗体 当婴儿粘膜免疫反应产生时，也可以保护牛奶 开发儿科疫苗以持续保护的机会。保护性抗体也可能是 作为治疗性抗体开发，可以治疗免疫功能低下的持续感染 患者，例如患有艾滋病毒/艾滋病的患者。我们希望鉴定240-400个免疫反应性隐孢子虫蛋白， 至少有3个将增加发病率较低的婴儿母亲的IgA水平 腹泻的比率或保护，以及婴儿适应性免疫的粪便IgA。此赠款申请 通过为疫苗奠定基础疫苗的基础 针对保护相关的特定粘膜IgA反应的研究。