Peptide-Protein Conjugate Vaccines

肽-蛋白结合疫苗

• 批准号: 7594238
• 负责人: rachel schneerson
• 金额: $ 65.99万
• 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/7594238
• 关键词: Adipic Acids; Adjuvant; Adsorption; Adverse effects; Amides; Animals; Anthrax Vaccines; Antibodies; Antibody Formation; Antigens; Bacillus anthracis; Binding; Bioterrorism; Carrier Proteins; Cessation of life; Child; Childhood; Conjugate Vaccines; Cytosol; Department of Defense; Development; Disease; Dose; Drug Formulations; Enzyme-Linked Immunosorbent Assay; Falciparum Malaria; Formaldehyde; Glutamic Acid; Goals; Human; Hydrazones; Hydrolysis; IgG1; IgG3; Immune Sera; Immunity; Immunization; Immunoglobulin G; Individual; Infection; Injection of therapeutic agent; Licensing; Malaria Vaccines; Mammalian Cell; Measures; Melanocytic nevus; Methods; Minor; Mole the mammal; Molecular Weight; Morbidity - disease rate; Mus; N-4-azido-2-nitrophenylpyridoxyl-5-phosphate; Nature; Numbers; Oximes; Pan Genus; Pan troglodytes; Peptides; Plasmodium falciparum; Proteins; Purpose; Rate; Reaction; Recombinants; Recruitment Activity; Reproduction spores; Scheme; Serum; Site; Sporozoites; Testing; Tetanus Toxoid; Time; Toxin; Vaccines; Virulence Factors; Week; aluminum sulfate; anthrax lethal factor; anthrax toxin; anti-IgG; base; capsule; circumsporozoite; circumsporozoite protein; dosage; edema factor; humanized monoclonal antibodies; immunogenic; immunogenicity; improved; mortality; peptide B; protective effect; repository; research clinical testing; research study; response; synthetic peptide; thioether; transmission process; volunteer

Bacillus anthracis: A recombinant PA was isolated from an uncapsulated strain. Several formaldehyde treated and or alum adsorbed formulations were immunogenic in mice. Clinical evaluation of these formulations is underway. 150 18-45 year old volunteers have been injected up to 4 times with no serious adverse effects. Local and systemic reactions were rare and minor. Serum IgG anti PA was measured in 246 sera of recruits injected with the Anthrax Vaccine Adsorbed (AVA ) stored at the Department of Defense Serum Repository. Paired sera were analyzed by ELISA. Serum conversion rates of ≥ 4-fold increase in antibody levels were: pre-post 3rd 85.3%, pre 4th-post 4th 67.9% and pre 6th-post 6th 45%. Geometric mean levels of all individuals were 59.9 mcg/mL following the 3rd injection, 157.4 mcg/mL following the 4th and 277microgram/mL following the 6th. The capsule has been isolated from a non toxic strain and it or corresponding synthetic peptides were bound to BSA, rEPA, rPA or tetanus toxoid. Additional conjugating methods using thioether, hydrazone and oxime linkages between the PGA and the proteins, with active groups at the C or N termini yielded conjugates immunogenic in mice, with no statistical difference between them. The induced antibodies were opsonophagocytic. Peptides 10 to 20-mer long, and 10-15 mole PGA per mole protein were the most immunogenic. Dose response experiments of an rPA-PGA conjugate, using between 0.31 to 20 mcg PGA/mouse showed 1.25 mcg to be optimal for a PGA response, while PA antibody levels increased with higher immunizing dosages. The use of alum increased PA antibody levels while having little effect upon anti PGA levels. Chimpanzees were immunized, 10 mcg PGA/animal, sc, one with PA-PGA another with TT-PGA, with the goal of preparing humanized monoclonal antibodies to PGA. Both chimps responded with antibodies to both vaccine components. Higher anti PGA levels were obtained in the TT-PGA injected chimp. PGS specific IgG1 and IgG3 were prepared. Plasmodium falciparum: The most studied experimental malaria vaccine candidate is the circumsporozoite protein (CSP), expressed extracellularly on the sporozoite, and various forms of its synthesized repeat unit, NANP. These vaccines were safe and immunogenic but poorly protective, even when administered with adjuvants. Based on our studies with peptides of the B. anthracis capsule, peptides of 4 repeat units of NANP were synthesized and bound to carrier proteins and their immunogenicity studied in general purpose mice without adjuvants by a scheme suitable for humans. High levels of antibodies were induced, with circumsporozoite neutralizing activity correlated roughly to levels measured by ELISA. In another approach, to provide a transmission blocking vaccine, Pfs25, a low molecular weight protein, non immunogenic by itself, was bound to itself or to carrier proteins by several methods: amide, hydrazone or thioether linkages and injected into mice to evaluate their antibody responses. All conjugates were immunogenic with booster responses upon reinjection. Remarkably, the serum antibody levels 3 and 7 months after immunization were higher than 1 week after the last injection. The best immunogens were created using adipic acid dihydrazide as the linker. Adsorption of the conjugates onto alum increased further the antibody levels. Transmission blocking activity of immune sera correlated with antibody levels measured by ELISA. The Pfs25 conjugate was also bound to a synthetic (NANP)5 of the repeat unit of the CSP. This conjugate induced IgG antibodies to both its components and adsorption onto alum increased further both levels: from 0.8 to 87g/mL for Pfs25 and from 3.3 to 53 EU/mL for CSP after 2 injections. Three months after the 2nd injection IgG anti Pfs25 increased to 524 mcg/mL and anti CSP to 120 EU/mL.

炭疽杆菌： 从未包囊化的菌株中分离重组PA。几种甲醛处理和/或明矾吸附制剂在小鼠中具有免疫原性。这些制剂的临床评价正在进行中。150名18-45岁的志愿者注射了4次，没有严重的不良反应。局部和全身反应罕见且轻微。检测了246名注射了储存在国防部血清储存库的吸附炭疽疫苗（AVA）的新兵血清中的血清IgG抗PA。通过ELISA分析配对血清。抗体水平4倍增加的血清转换率为：前3天后85.3%，前4天后67.9%，前6天后45%。第3次注射后，所有个体的几何平均水平为59.9 mcg/mL，第4次注射后为157.4 mcg/mL，第6次注射后为277微克/mL。该胶囊已从无毒菌株中分离，并且其或相应的合成肽与BSA、rEPA、rPA或破伤风类毒素结合。在PGA和蛋白质之间使用硫醚、腙和肟键的其他缀合方法，在C或N末端具有活性基团，在小鼠中产生免疫原性缀合物，它们之间没有统计学差异。诱导的抗体具有调理吞噬作用。肽10至20聚体长，和10-15摩尔PGA每摩尔蛋白质是最免疫原性。使用0.31至20 mcg PGA/小鼠的rPA-PGA偶联物的剂量反应实验表明，1.25 mcg对于PGA反应是最佳的，而PA抗体水平随着免疫剂量的增加而增加。明矾的使用增加PA抗体水平，而对抗PGA水平几乎没有影响。 免疫黑猩猩，10 mcg PGA/动物，sc，一个用PA-PGA，另一个用TT-PGA，目的是制备针对PGA的人源化单克隆抗体。两只黑猩猩都对疫苗的两种成分产生了抗体。在TT-PGA注射的黑猩猩中获得更高的抗PGA水平。制备PGS特异性IgG 1和IgG 3。 恶性疟原虫： 研究最多的实验性疟疾疫苗候选物是环子孢子蛋白（CSP），其在子孢子上细胞外表达，以及其合成的重复单元NANP的各种形式。这些疫苗是安全和免疫原性的，但保护性差，即使与佐剂一起施用。基于我们对B肽的研究。炭疽菌囊膜中，NANP的4个重复单元的肽被合成并结合到载体蛋白上，并通过适合于人类的方案在无佐剂的通用小鼠中研究其免疫原性。诱导高水平的抗体，环子孢子中和活性大致与ELISA测定的水平相关。在另一种方法中，为了提供传递阻断疫苗，Pfs 25（一种本身无免疫原性的低分子量蛋白质）通过几种方法（酰胺、腙或硫醚键）与自身或载体蛋白结合，并注射到小鼠中以评价其抗体应答。所有缀合物都是免疫原性的，在重新注射后具有加强应答。值得注意的是，免疫后3个月和7个月的血清抗体水平高于最后一次注射后1周。使用己二酸二酰肼作为接头产生最佳免疫原。结合物吸附到明矾上进一步增加了抗体水平。免疫血清的传递阻断活性与通过ELISA测量的抗体水平相关。 Pfs 25缀合物还结合至CSP的重复单元的合成（NANP）5。该缀合物诱导针对其两种成分的IgG抗体，并且在明矾上的吸附进一步增加了两种水平：注射2次后，Pfs 25从0.8至87 g/mL，CSP从3.3至53 EU/mL。第2次注射后3个月，IgG抗Pfs 25增加至524 mcg/mL，抗CSP增加至120 EU/mL。