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: 6840685
• 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/6840685
• 关键词: Bacillus; Borrelia; China; Neisseria meningitidis; Neisseria meningitidis vaccine; Shigella; Shigella vaccines; Streptococcus infection; Streptococcus pneumoniae; Streptococcus vaccine; active immunization; anthrax vaccines; bacterial polysaccharides; bacterial vaccines; bactericidal immunity; bioterrorism /chemical warfare; clinical trial phase III; cross immunity; drug screening /evaluation; human subject; human therapy evaluation; immunoconjugates; laboratory mouse; malaria vaccines; patient oriented research; 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 substrates 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 ourpreparation 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，包括脂多糖（LPS）的荚膜多糖（CPS）或O-特异性多糖（O-SP），既是必需的毒力因子，又是保护性抗原。CPS或O-SP与医学上有用的蛋白质共价结合以形成缀合物既增加了它们的免疫原性，又赋予这些抗体T细胞依赖性，使它们成为适合婴儿和儿童的疫苗。与重组无毒白喉毒素（CRM 9）结合的宋内志贺菌O-SP和与重组无毒琥珀酰化外蛋白A（rEPA-succ）结合的福氏2a志贺菌O-SP是安全的，并在1-4岁儿童中诱导了针对同源LPS的IgG抗体：福氏2a志贺菌偶联物在92%的儿童中诱导了4倍升高，宋内志贺菌偶联物在85%中诱导了4倍升高。针对1-4岁儿童的这些结合物的第三阶段研究已经开始。目前正在与中国河南省兰州疫苗研究所和省医疗中心合作，计划对这些疫苗进行临床试验。 交叉反应多糖，B.双歧杆菌：为了研究与同源CPS同时沿着施用交叉反应的CPS是否比单独使用同源CPS具有优势，分离了短小芽孢杆菌Sh 18的细胞壁多糖（PS），其被报道与流感嗜血杆菌B型（Hi B）的CPS交叉反应，并使用GC-MS、NMR、NMR、MS和MS研究了其结构。快速原子轰击和几种糖降解技术。它是由聚核糖醇磷酸、C2位被N-乙酰氨基葡萄糖取代的聚核糖醇磷酸和聚甘油磷酸组成。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小鼠中评价其免疫原性。缀合物诱导的抗体与同源的和几个交叉反应的多糖反应。 A群脑膜炎奈瑟菌引起地方性和流行性脑膜炎，特别是在非洲脑膜炎带。有效和可用的CPS疫苗未得到充分利用。为了进一步提高其免疫原性，使用ADH作为首先与CPS或BSA结合的接头将其缀合至BSA。与单独的CPS相反，缀合物在小鼠中具有免疫原性，在第2次和第3次注射后具有加强应答，并且杀菌活性与IgG抗CPS水平相关。交叉反应性多糖，大肠杆菌K93和短小B杆菌SH 17的共轭物，没有诱导抗Men。皇家检察署伯氏疏螺旋体是莱姆病的病原体，通过感染的硬蜱叮咬传播。目前已有一种蛋白质疫苗，但对12岁以下儿童无效。LPS在其他螺旋体中也有描述，但它存在于B中。Burgdorferi一直在争论。我们还不能确认它的存在。对LPS的搜索揭示了2种独特的糖脂：胆固醇基6-O-棕榈酰-β-D-吡喃半乳糖苷（BBGL 1）和1-乳棕榈酰-2-O-油基-3-O-α-D吡喃半乳糖基-sn-甘油（BBGL 2）。有证据表明它们是表面暴露的。将BBGL 1以各种制剂注射到小鼠中诱导特异性抗体，按诱导水平的顺序为：CFA、PBS、DMSO、角鲨烯。抗体的生物效应正在被消化。分离脑膜炎奈瑟氏菌Gr.W135的CPS并将其与金黄色葡萄球菌重组超抗原2缀合，该CPS是最近在非洲爆发的原因。 炭疽杆菌是人类致死性感染的潜在原因，具有2个基本毒力因子，如果没有其中任何一个，则对人类没有致病性。这些因素是：1.炭疽毒素，2.胶囊。该毒素由3种肽组成：致死因子，水肿因子和保护性抗原，每种本身无毒。PA是与哺乳动物细胞结合的毒素部分。它必须有一个20 KDa的肽水解，暴露出LF或EF可能结合的位点，从而产生毒素，酶促修饰哺乳动物细胞胞质溶胶中的底物。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。许可的疫苗是安全和保护性的，但有限制，证明改进疫苗的发展。从发酵罐中生长的未包封菌株中分离出重组PA。发现几种含有甲醛处理和明矾吸附材料的制剂在小鼠中具有免疫原性。正在制备临床批次。从一株无毒菌株中分离到一种荚膜，并将其或相应的合成肽与BSA、rEPA或rPA结合。为了鉴定不同长度的最佳构建体肽，将在C或n末端具有活性基团的D或L构型的5、10和20聚体缀合。通过物理化学和免疫学测定以及在5-6周龄小鼠中的免疫原性来表征缀合物。与单独的PGA相反，所有缀合物都具有免疫原性，并且D-PGA诱导的抗体具有调理吞噬作用。rPA是更有效的载体。 肺炎链球菌6 B型是最常见和免疫原性最低的类型之一，与破伤风类毒素结合，并在慢性阻塞性肺病患者中进行评价，与这些患者中的23价多糖疫苗以及健康年轻人的反应进行比较。3组间抗体水平无统计学差异。调理吞噬作用与IgG抗6 B型水平有关。 疟原虫：我们在制备合成S. 1型和B型。炭疽实验疫苗正应用于该生物体的受精卵蛋白，试图增加其免疫原性。