Protein and Polysaccharide Conjugate Vaccines to enteric diseases

肠道疾病蛋白质和多糖结合疫苗

• 批准号: 7594170
• 负责人: SHOUSUN C SZU
• 金额: $ 39.12万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7594170
• 关键词: 5 year old; Accounting; Adult; Adverse reactions; Africa; Amino Acids; Antibodies; Antigens; Bacteria; Bacterial Toxins; Binding; Campylobacter jejuni; Capsid Proteins; Carrier Proteins; Cells; Cessation of life; Child; Childhood; Cholera; Collaborations; Conjugate Vaccines; Developed Countries; Developing Countries; Developmental Therapeutics Program; Diarrhea; Disease; Dose; Enteral; Escherichia coli; Escherichia coli K12; Escherichia coli O157; Exotoxins; Formalin; Gangliosides; Genes; Goals; Guillain-Barré Syndrome; Haemophilus influenza type b polysaccharide vaccine-tetanus toxin conjugate; Health; Heating; Hemolytic-Uremic Syndrome; Human; Immune Sera; Immune response; Immunoglobulin G; Infant; Infantile Diarrhea; Infection; Injection of therapeutic agent; Intussusception; Israel; Length; Licensing; Lipopolysaccharides; Marketing; Medical center; Mus; O Antigens; Patients; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Phase III Clinical Trials; Polysaccharides; Principal Investigator; Proteins; Pseudomonas aeruginosa; Recombinants; Reporting; Rotavirus; Rotavirus Infections; Rotavirus Vaccines; Salmonella; Salmonella paratyphi; Salmonella typhi; Serotyping; Serum; Shiga Toxin; Sialic Acids; Site; Surface; Swelling; Teenagers; Toddler; Toxin; Toxoids; Typhoid Fever; Typhoid Vaccine; Vaccines; Vi capsular polysaccharide; Vibrio cholerae; Vibrio cholerae O1; Viral; Virulence Factors; Virus; Whole Cell Vaccine; anti-IgG; bactericide; base; capsule; chemical synthesis; design; dosage; enterotoxigenic Escherichia coli; field study; follow-up; immunogenic; immunogenicity; improved; lipooligosaccharide; mutant; neutralizing antibody; oral vaccine; pathogen; polypeptide; programs; 5 year old; Accounting; Adult; Adverse reactions; Africa; Amino Acids; Antibodies; Antigens; Bacteria; Bacterial Toxins; Binding; Campylobacter jejuni; Capsid Proteins; Carrier Proteins; Cells; Cessation of life; Child; Childhood; Cholera; Collaborations; Conjugate Vaccines; Developed Countries; Developing Countries; Developmental Therapeutics Program; Diarrhea; Disease; Dose; Enteral; Escherichia coli; Escherichia coli K12; Escherichia coli O157; Exotoxins; Formalin; Gangliosides; Genes; Goals; Guillain-Barré Syndrome; Haemophilus influenza type b polysaccharide vaccine-tetanus toxin conjugate; Health; Heating; Hemolytic-Uremic Syndrome; Human; Immune Sera; Immune response; Immunoglobulin G; Infant; Infantile Diarrhea; Infection; Injection of therapeutic agent; Intussusception; Israel; Length; Licensing; Lipopolysaccharides; Marketing; Medical center; Mus; O Antigens; Patients; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Phase III Clinical Trials; Polysaccharides; Principal Investigator; Proteins; Pseudomonas aeruginosa; Recombinants; Reporting; Rotavirus; Rotavirus Infections; Rotavirus Vaccines; Salmonella; Salmonella paratyphi; Salmonella typhi; Serotyping; Serum; Shiga Toxin; Sialic Acids; Site; Surface; Swelling; Teenagers; Toddler; Toxin; Toxoids; Typhoid Fever; Typhoid Vaccine; Vaccines; Vi capsular polysaccharide; Vibrio cholerae; Vibrio cholerae O1; Viral; Virulence Factors; Virus; Whole Cell Vaccine; anti-IgG; bactericide; base; capsule; chemical synthesis; design; dosage; enterotoxigenic Escherichia coli; field study; follow-up; immunogenic; immunogenicity; improved; lipooligosaccharide; mutant; neutralizing antibody; oral vaccine; pathogen; polypeptide; programs

Surface polysaccharides of Gram-negative pathogens, capsules or lipopolysaccharides, are essential virulence factors and protective antigens. Lipopolysaccharides need to be detoxified and their O-specific polysaccharides (O-SP) isolated. Immunogenicity of polysaccharides can be improved by binding to carrier proteins. Bacterial toxins or toxoids and viral capsid proteins may be protective antigens and can also serve as carrier proteins. Salmonella typhi. (Co-principal Investigator, Feng-Ying Lin). The Vi capsular polysaccharide of S. typhi is a licensed typhoid vaccine. To improve its immunogenicity in young children, Vi was conjugated to a recombinant exoprotein A of Pseudomonas aeruginosa (rEPA). A phase 3 trial of the Vi-rEPA in 11,600 Vietnamese 2-5-year olds showed an efficacy of 89% at 47 months. A study of various dosages for 2-to-5-year olds showed a higher dosage was more immunogenic. A phase 2 trial in 301 infants started in 2006. Vi-rEPA is injected concurrently with DPT at 2, 4, 6 and 12 months. Controls receive Hib-TT +DTP or DTP. IgG anti-Vi levels will be compared with those elicited in the phase 3 trial. A 10 years follow up of adults injected once with Vi-rEPA in the phase 1 study, showed a G.M. IgG anti-Vi level20 fold higher than the proposed protective level (P<0.0001). The duration of serum anti-Vi IgG in children immunized with Vi-rEPA 8 years ago is under analysis and will be compared with levels 4 years ago and of those of unimmunized controls in another village. Salmonella paratyphi A (SPA) is the second most common cause of enteric fever in developing countries. In our phase 1 and 2 studies, S. paratyphi A O-SP conjugated to TT was safe and immunogenic in adults, teenagers, and toddlers. A mutant S. paratyphi A was constructed by replacing the LPS elongation chain length regulator wzz gene with that of E. coli K12. A conjugate vaccine of this elongated O-SP was prepared and its immunogenicity will be evaluated in mice. E. coli O157 is a major cause of hemolytic uremic syndrome, especially in young children. In our phase 2 trial of O-SP-rEPA conjugate in 2-5 year-olds (at Carolina Medical Center), the vaccine was shown to be safe, immunogenic and elicited bactericidal antibodies; 98% had 4 fold rise of IgG anti-LPS at 6 months. A major virulence factor of E. coli O157 is the Shiga toxin II. A mutant toxoid constructed by Alison OBrien conjugated with O-SP for enhanced protection is under study. Enterotoxigenic E. coli (ETEC): ETEC is the most common cause of diarrhea in developing countries. ETEC secretes two exotoxins: heat-labile toxin (LT) and heat stable toxin (ST), a polypeptide of 19 amino acids. LT is immunogenic in humans. However its efficacy against infection has not been demonstrated. ST is small, non-immunogenic and difficult to purify. A non-toxic mutant, rLT, from John Clements was formalin-treated and in mice it induced high levels of IgG anti-LT. There was a dose dependent swelling at the injection site. In collaboration with Donald Robertson, the ST was purified, to be suitable for conjugation with LT or other proteins. Vibrio cholerae O1 remains a major health problem in the Indian subcontinent and in Africa. Field studies showed that the vibriocidal activity of antisera is directed towards the LPS. In our Phase 1 trial the O-SP conjugates elicited high levels of IgG anti-LPS with vibriocidal activity. Chemical synthesis of O1 V. cholera O-SP is underway. Campylobacter jejuni infection is common and may cause serious complications such as Guillain- Barre syndrome; possibly due to the structural similarity between gangliosides and lipooligosaccharides (LOS) of C. jejuni. Protein conjugates of type 2 LOS or of the de-acylated LOS elicited anti-LOS IgG and bactericidal antibodies. 28 isolates of C. jejuni from pediatric patients in Israel were analyzed for serotype distribution; a diverse serotype distribution was found. Approximately 40% of the 16 isolates contain sialic acid in their LOS and bind to human ganglioside antisera. Rotavirus infection is the most common cause of infantile diarrhea worldwide. Current vaccines are reassortant whole-cell oral vaccines. One such vaccine, Rotashield, was withdrawn from the market after a suspected increase in intussusception in its recipients. We designed parenteral vaccines based on capsid proteins VP8 and VP7. Mice injected with conjugate capsid proteins had neutralizing antibodies against rotavirus of homologous (P4) and heterologous (P8) serotypes.

革兰氏阴性病原体，胶囊或脂多糖的表面多糖是基本的毒力因子和保护性抗原。需要对脂多糖进行解毒，并分离其O特异性多糖（O-SP）。通过与载体蛋白结合，可以改善多糖的免疫原性。细菌毒素或毒素和病毒衣壳蛋白可能是保护性抗原，也可以用作载体蛋白。 沙门氏菌Typhi。 （联合首席研究员，冯林林）。 Typhi S. typhi的VI胶囊多糖是持牌伤寒疫苗。为了改善其在幼儿中的免疫原性，VI与铜绿假单胞菌（REPA）的重组异蛋白A结合在一起。 VI-REPA的3阶段试验在11,600名越南2-5岁的年轻人中显示出47个月时的疗效89％。对2至5岁年龄段的各种剂量的研究表明，较高的剂量更为免疫原性。在2006年开始了301例婴儿的2期试验。VI-REPA与DPT同时注射2、4、6和12个月。控件接收HIB-TT +DTP或DTP。 IgG抗VI水平将与第三阶段试验中引起的抗VI水平进行比较。 在第1阶段研究中对VI-REPA注射一次的成年人的10年随访显示了G.M. IgG抗VI水平20倍高于提议的保护水平（P <0.0001）。 8年前，用VI-REPA免疫的儿童血清抗VI IgG的持续时间正在分析，并将与4年前的水平以及另一个村庄的无免疫对照的水平进行比较。 沙门氏菌A（SPA）是发展中国家肠发烧的第二大最常见原因。在我们的第1阶段和第2阶段的研究中，在成年人，青少年和学步儿童中，与TT结合的副链球菌A O-SP是安全的和免疫原性的。通过用大肠杆菌K12代替LPS伸长链长度调节基因WZZ基因来构建突变链球菌A。制备了该拉长O-SP的共轭疫苗，并将在小鼠中评估其免疫原性。 大肠杆菌O157是溶血性尿毒症综合征的主要原因，尤其是在幼儿中。在2-5岁（在卡罗来纳州医疗中心）中O-SP-REPA结合物的2期试验中，该疫苗被证明是安全，免疫原性和引起的杀菌抗体的；在6个月时，98％的IgG抗LP升高了4倍。大肠杆菌O157的主要毒力因子是志贺毒素II。正在研究由Alison Obrien与O-SP结合以增强保护的突变体毒素。 肠毒素大肠杆菌（ETEC）：ETEC是发展中国家腹泻的最常见原因。 ETEC分泌两种外毒素：热能毒素毒素（LT）和热稳定毒素（ST），这是19个氨基酸的多肽。 LT在人类中具有免疫原性。但是，尚未证明其对感染的功效。 ST很小，不发育，难以纯化。 约翰·克莱门茨（John Clements）的无毒突变体RLT经过福尔马林处理，在小鼠中，它诱导了高水平的IgG抗LT。注射部位有剂量依赖性肿胀。与唐纳德·罗伯逊（Donald Robertson）合作，ST被纯化，适合与LT或其他蛋白质结合。 在印度次大陆和非洲，Vibrio Cholerae O1仍然是一个主要的健康问题。现场研究表明，抗血清的颤动活性针对LPS。在我们的第1阶段试验中，O-SP偶联会引起高水平的IgG抗LP，具有颤动活性。 O1 V.霍​​乱O-SP的化学合成正在进行中。 弯曲杆菌的空肠感染很常见，可能会引起严重的并发症，例如Guillain-Barre综合征。可能是由于Jejuni的神经节苷脂和脂肪糖（LOS）之间的结构相似性。 2型LOS的蛋白质结合物或去酰化LOS的抗LOS IgG和杀菌抗体的蛋白结合物。分析了以色列儿科患者的28种梭状芽孢杆菌的血清型分布；发现了各种血清型分布。在16个分离株中，大约有40％的LOS含有唾液酸，并与人神经节苷脂抗血清结合。 轮状病毒感染是全球婴儿腹泻的最常见原因。当前的疫苗是重新排育的全细胞口服疫苗。一种这样的疫苗Rotashield被怀疑是其接受者的肠suse脚增加后被撤回了市场。我们设计了基于衣壳蛋白VP8和VP7的肠胃外疫苗。注射结合胶囊蛋白的小鼠具有对同源（P4）和异源（P8）血清型的轮状病毒的抗体中和抗体。