Human Immune Response To Polysaccharide-protein Conjugat

人体对多糖-蛋白质缀合物的免疫反应

• 批准号: 6992836
• 负责人: RACHEL SCHNEERSON
• 金额: --
• 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/6992836
• 关键词: Bacillus; Bacillus anthracis; Borrelia; China; Neisseria meningitidis; Plasmodium falciparum; Shigella; Shigella vaccines; Streptococcus pneumoniae; active immunization; bacterial polysaccharides; bacterial vaccines; bactericidal immunity; biotechnology; bioterrorism /chemical warfare; clinical trial phase III; cross immunity; human subject; human therapy evaluation; immune response; immunoconjugates; laboratory mouse; malaria vaccines; patient oriented research; preschool child (1-5); synthetic vaccines; vaccine development; vaccine evaluation

SHIGELLA: Surface polysaccharides of pathogenic bacteria, including capsular polysaccharides (CPS) or the O-specifc polysaccharide (O-SP) of lipopolysaccharides (LPS),serve both as essential virulence factors and as protective antigens. Covalent binding of CPS or of O-SP to medically-useful proteins to form conjugates both increases their immunogenicity and confers T-cell dependence to these saccharides making them suitable vaccines for infants and children. The O-SP of Shigella sonnei bound to recombinant non toxic diphtheria toxin (CRM9)and of S.flexneri 2a bound to the recombinant, non toxic, succinylated, exoprotein A (rEPA-succ) were safe and induced IgG antibodies to the homologous LPS in 1-4 year-olds: a 4-fold rise was induced by the S.flexneri 2a conjugate in 92% of the children, the S.sonnei conjugate in 85%. A phase 3 study of these conjugates in 1-4 year-olds has begun. In collaboration with the Lanzhou Vaccine Institute and Provincial Medical Center in Henan, China, a clinical trial of these is being planned. CROSS REACTING POLYSACCHARIDES,B.PUMILUS: To investigate if concurrent administration of a cross-reacting along with a homologous CPS has an advantage over the use of the homologous CPS alone, the cell wall polysaccharide (PS) of Bacillus pumilus, Sh18, reported to cross react with the CPS of haemophilus influenzae type b (Hib), was isolated and it's structure investigated using GC-MS,NMR, fast atom bombardment and several sugar degrading techniques. It was shown to be composed of polyribitolphosphate, polyribitolphsphate substituted at C2 with N-acetyl glucosamine and polyglycerolphosphate. Besides with the anti Hib it reacted with anti Staphylococcus epidermidis. The polysaccharide was conjugated to carrier proteins and it's immunogenicity evaluated in GP mice. Conjugate-induced antibodies reacted with the homologous and several cross-reacting polysaccharides. NEISSERIA MENINGITIDIS group A causes endemic and epidemic meningitis, notably in the meningitis belt of Africa. A CPS vaccine , effective and available, is underutilized. To further improve it's immunogenicity it was conjugated to BSA using ADH as a linker bound first to the CPS or the BSA. Contrary to the CPS alone the conjugates were immunogenic in mice, with booster responses after 2nd and 3rd injections and bactericidal activity related to IgG anti CPS levels. Conjugates of the cross reactive polysaccharides, e.coli K93 and B.pumilus SH17,did not induce anti Men. A CPS. BORRELIA BURGDORFERI, a spirochete transmitted though the bite of infected Ixodes ticks, is the etiologic agent of Lyme disease. A protein vaccine against it is available but is not effective below the age of 12 years. LPS has been described in other spirochetes but it's presence in B. burgdorferi has been debated. We have not been able to confirm it's presence. The search for LPS revealed 2 unique glycolipids:cholesteryl 6-O-palmitoyl-beta-D-galactopyranoside (BBGL1) and1-Opalmitoyl-2-O-oleyl-3-O-alpha-D galactopyranosl-sn-glycerol (BBGL2) There is evidence that they are surface exposed. Injected in various formulations into mice BBGL1 induced specific antibodies,in order of induced levels: CFA,PBS,DMSO,squalene. The biological effect of the antibodies is being ingestigated. The CPS of N.meningitidis Gr.W135, responsible for recent outbreaks in Africa, was isolated and conjugated to S.aureus recombinant super antigen 2. BACILLUS ANTHRACIS, a potential cause of lethal human infection, has 2 essential virulence factors without either of which it is not pathogenic for humans. These factors are: 1.anthrax toxin, 2.a capsule. The toxin is composed of 3 peptides: Lethal Factor, Edema Factor and Protective Antigen, each by itself non toxic. PA is the toxin part that binds to mammalian cells. It has to have a 20 KDa peptide hydrolyzed off exposing a site to which LF or EF may bind rendering toxins that enzymatically modify substrattes in mammalian cell cytosol. The capsule is composed of poly-D-gamma-glutamic acid(PGA).It is non-immunogenic and it's protective effect unknown. The licensed vaccine is safe and protective but has limitations that justify development of improved vaccines. A recombinant PA was isolated from an unencapsulated strain grown in a fermenter. Several formulations with formaldehyde treated and alum adsorbed materials were found to be immunogenic in mice. Clinical lots are being prepared. The capsule has been isolated from a non toxic strain and it or corresponding syntethic peptideswere bound to BSA, rEPA or rPA. To identify the optimal construct peptides of varying lengths, 5,10 and 20-mers,of D or L configuration with active groups at the C or n termini were conjugated. the conjugates were characterizedbby physico-chemical and immunological assays and immunogenicity in 5-6 week old mice. Opposed to PGA alone all conjugates were immunogenic and the D-PGA -induced antibodies were opsonophagocytic. rPA was the more effective carrier. STREPTOCOCCUS PNEUMONIAE type 6B,one of the most common and least immunogenic types, was conjugated to tetanus toxoid and evaluated in patients with chronic obstructive pulmonary disease, compared to the 23-valent polysacharide vaccine in these patients, and to the response of healthy young adults. No statistical differences in antibody levels were found between the 3 groups. Opsonophagocytosis was related to IgG anti type 6B levels. PLASMODIUM FALCIPARUM:Techniques used in our preparation of our synthetic S. dysenteriae type 1 and B. anthracis experimental vaccines are being applied to proteins of the zygote of this organism, in an attempt to increase their immunogenicity.

志贺氏菌：病原菌的表面多糖，包括荚膜多糖(CPS)或脂多糖(LPS)的O-特异性多糖(O-SP)，既是基本的毒力因子，也是保护性抗原。CPS或O-SP与医学上有用的蛋白质共价结合，形成偶联物，既增加了它们的免疫原性，又赋予这些糖T细胞依赖性，使它们成为适合婴儿和儿童的疫苗。结合重组无毒白喉毒素(CRM9)的宋内氏志贺氏菌O-SP和结合重组无毒琥珀酸化外蛋白A的福氏2a志贺菌(REPA-Succ)在1~4岁儿童中均能安全地诱导抗同源内毒素的抗体：福氏志贺氏菌2a结合物可使92%的儿童产生4倍以上的抗体，宋内氏志贺菌结合物为85%。这些结合物在1-4岁儿童中的第三阶段研究已经开始。与兰州疫苗研究所和河南省医学中心中国合作，正在计划对这些药物进行临床试验。 交叉反应多糖，B.PUMILUS：为了研究交叉反应与同源CPS同时给予是否比单独使用同源CPS更有优势，分离了短小芽孢杆菌Sh18与b型流感嗜血杆菌CPS交叉反应的细胞壁多糖，并用GC-MS、核磁共振、快原子轰击和几种糖降解技术对其结构进行了研究。结果表明，它是由多聚核糖醇磷酸、C2位N-乙酰氨基葡萄糖取代的多聚核糖醇磷酸盐和聚甘油磷酸组成。除与抗Hib反应外，还与抗表皮葡萄球菌反应。将该多糖与载体蛋白偶联，并在GP小鼠体内检测其免疫原性。结合物诱导的抗体与同源和几种交叉反应的多糖反应。 A组脑膜炎奈瑟菌引起地方性和流行性脑膜炎，特别是在非洲的脑膜炎地带。一种有效和可用的CPS疫苗没有得到充分利用。为了进一步提高其免疫原性，将其与牛血清白蛋白偶联，用ADH作为连接物，首先与CPS或BSA结合。与单独的CPS相反，结合物在小鼠体内具有免疫原性，在第二次和第三次注射后具有增强反应，杀菌活性与Ig G抗CPS水平有关。交叉反应多糖的偶联物E.coliK93和B.puilus SH17不能诱导抗人类抗体。一个CPS。伯氏疏螺旋体是莱姆病的病原体，是一种通过受感染的硬蜱叮咬传播的螺旋体。目前有一种针对这种疾病的蛋白质疫苗，但在12岁以下无效。脂多糖曾在其他螺旋体中被描述过，但它在伯氏杆菌中的存在一直存在争议。我们还不能确认它的存在。对脂多糖的搜索发现了两种独特的糖脂：胆固醇6-O-棕榈酰基-β-D-半乳糖苷(BBGL1)和1-乳糖基-2-O-油基-3-O-α-D-半乳糖基-sn-甘油(BBGL2)。有证据表明它们是表面暴露的。以不同剂型注射小鼠BBGL1诱导产生特异性抗体，诱导水平依次为：CFA、PBS、DMSO、角鲨烯。这些抗体的生物学效应正在调查中。分离了最近在非洲暴发的脑膜炎奈瑟氏菌GRW135的CPS，并将其与金黄色葡萄球菌重组超级抗原2结合。 炭疽杆菌是人类致命感染的潜在原因，它有两个基本的毒力因子，如果没有这两个因子，它就不会对人类产生致病作用。这些因素是：1.炭疽毒素；2.胶囊。该毒素由致死因子、水肿素和保护性抗原3种多肽组成，每种多肽本身均无毒。PA是与哺乳动物细胞结合的毒素部分。它必须有一个20 KDa的多肽被水解，暴露出一个Lf或EF可能与之结合的位置，从而产生毒素，对哺乳动物细胞胞浆中的底物进行酶促修饰。该胶囊由聚D-γ-谷氨酸(PGA)组成，无免疫原性，其保护作用尚不清楚。获得许可的疫苗是安全和保护的，但也有局限性，证明有理由开发改进的疫苗。从发酵罐中培养的无囊化菌株中分离到一株重组PA。几种经甲醛处理和明矾吸附材料的配方在小鼠身上被发现具有免疫原性。临床批次正在准备中。该胶囊是从无毒菌株中分离出来的，它或相应的合体多肽与BSA、REPA或RPA结合。为了确定不同长度的最佳构建肽，将D或L构型的5，10和20-MERS与C或n末端具有活性基团的5，10和20-MERS偶联。用理化、免疫学方法和5-6周龄小鼠免疫原性对其进行了鉴定。与PGA单独作用相反，所有结合物都是免疫原性的，而D-PGA诱导的抗体是吞噬细胞的。RPA是更有效的载体。 肺炎链球菌6B型是最常见且免疫原性最低的类型之一，它与破伤风类毒素结合，并在慢性阻塞性肺疾病患者中进行评估，与23价多糖疫苗在这些患者中的比较，以及对健康年轻人的反应。3组间抗体水平差异无统计学意义。吞噬细胞功能与Ig G抗6B抗体水平相关。 恶性疟原虫：我们在制备合成1型痢疾杆菌和炭疽杆菌实验疫苗时使用的技术正在应用于这种生物受精卵的蛋白质，试图增加它们的免疫原性。