Haemophilus ducreyi and Bordetellae

博德特氏菌： 博德特氏菌是革兰氏阴性杆菌，引起哺乳动物和鸟类的呼吸道感染。临床上重要的是B。百日咳，B. B.副百日咳和B.支气管炎B。百日咳疫苗在预防婴儿和儿童百日咳方面取得了成功。抗B的兽用疫苗。支气管败血症，但其疗效和作用方式尚未确定。没有预防B的疫苗。副百日咳。基于血清IgG抗LPS可赋予对非荚膜革兰氏阴性菌的免疫力的概念，我们研究了B的O-特异性多糖（O-SP）的化学、血清学和免疫学性质。支气管败血症和B.通过不同的降解程序获得的副百日咳。一种类型的B。副百日咳和两种类型的B。支气管败血症O-SP的非还原性末端糖的结构被识别，两个B之间没有交叉反应。支气管败血症型。竞争性抑制试验表明，这些O-SP的非还原端的免疫优势。B的缀合物。支气管败血症和B.通过两种新的方法制备副百日咳O-SP：使用通过LPS的温和酸水解暴露的Kdo残基或通过LPS的脱氨基暴露的核心葡糖胺残基，用于结合氨氧基化蛋白。 两种偶联方法均在中性pH、室温和短时间内进行。所有的共轭物，作为盐水溶液注射在一个分数的估计人类剂量，诱导小鼠抗体的同源O-SP。这些方法可应用于制备基于LPS的疫苗对其他革兰氏阴性菌。 H. ducreyi： 本文对两株软下疳致病菌杜克雷嗜血杆菌的低脂多糖进行了研究。LOS的碳水化合物部分由分别以部分唾液酸化的乳糖胺或乳糖终止的九种或六种糖组成。用2.5 - 5 mcg来自任一菌株的纯化LOS重复皮下免疫小鼠诱导IgG应答，主要针对内核区域。仅检测到低水平的针对市售新乳四糖或唾液酸-新乳四糖的抗体，所述新乳四糖含有LOS的末端乳糖胺结构。通过LOS注射诱导的抗体不与具有末端（唾液酸）乳糖胺结构的人糖脂结合，但如通过免疫薄层色谱法测定的，它们与脂质A或脂质A-Kdo-Kdo β结合。这些血清与其同源细菌的结合较弱，杀菌能力较弱。新乳四糖或唾液酸-新乳四糖结合人血清白蛋白免疫小鼠产生低水平的抗LOS抗体。为了保留缀合物中LOS的整个糖部分，我们将存在于OS的末端还原端的3-脱氧-D-甘露-辛酮糖酸（Kdo）连接到与载体蛋白结合的双功能接头的氨氧基。这些缀合物诱导低水平的抗LOS，主要是内核，与同源细菌弱结合，并且具有低杀菌活性。

Bordetellae: Bordetellae are Gram-negative bacilli causing respiratory tract infections of mammals and birds. Clinically important are B. pertussis, B. parapertussis and B. bronchiseptica. B. pertussis vaccines have been successful in preventing pertussis in infants and children. Veterinary vaccines against B. bronchiseptica are available, but their efficacy and mode of action are not established. There is no vaccine against B. parapertussis. Based on the concept that immunity to non-capsulated Gram-negative bacteria may be conferred by serum IgG anti-LPS we studied chemical, serological and immunological properties of the O-specific polysaccharides (O-SP) of B. bronchiseptica and B. parapertussis obtained by different degradation procedures. One type of the B. parapertussis and two types of B. bronchiseptica O-SP were recognized based on the structure of their non-reducing end saccharide; no cross-reaction between the two B. bronchiseptica types was observed. Competitive inhibition assays showed the immunodominance of the non-reducing end of these O-SP. Conjugates of B. bronchiseptica and B. parapertussis O-SP were prepared by two new methods: using the Kdo residue exposed by mild acid hydrolysis of the LPS or the core glucosamine residue exposed by deamination of the LPS, for binding to an aminooxylated protein. Both coupling methods were carried out at a neutral pH, room temperature, and in a short time. All conjugates, injected as saline solutions at a fraction of an estimated human dose, induced antibodies in mice to the homologous O-SP. These methodologies can be applied to prepare LPS-based vaccines against other Gram-negative bacteria. H. ducreyi: Lipooligosaccharides (LOS) of two strains of Haemophilus ducreyi, the causative agent of chancroid, were studied. The carbohydrate part of the LOS consists of either nine or six sugars terminating with partially sialylated lactosamine or lactose, respectively. Repeated subcutaneous immunization of mice with 2.5 - 5 mcg of purified LOS from either strain induced IgG responses, mostly to the inner core region. Only low levels of antibodies to the commercially obtained neolactotetraose or sialyl-neolactotetraose, containing the terminal lactosamine structure of the LOS, were detected. Antibodies induced by LOS injection did not bind to human glycolipids with terminal (sialyl)lactosamine structures, but they bound to Lipid A or Lipid A-Kdo-Kdo saccharides as assayed by immune thin-layer chromatography. These sera bound weakly to their homologous bacteria and were weakly bactericidal. Mice immunized with neolactotetraose or sialyl-neolactotetraose conjugated to human serum albumin produced low levels anti-LOS antibodies. To preserve the entire saccharide part of LOS in the conjugate we linked the 3-deoxy-D-manno-octulosonic acid (Kdo) present on the terminal reducing end of the OS to an amiooxy group of a bifunctional linker bound to carrier protein. These conjugates induced low levels of anti-LOS, mostly to the inner core, bound weakly to the homologous bacteria and had low bactericidal activity.