Bordetellae and Haemophilus ducreyi

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的非还原性末端糖结构，两种B无交叉反应。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来制备副百日咳O-SP，以结合氨氧基化蛋白。 两种偶联方法均在中性pH、室温和短时间内进行。所有偶联物以生理盐水溶液形式注射到小鼠体内，剂量仅为估计人体剂量的一部分，均诱导产生针对同源O-SP的抗体。这些方法可用于制备针对其他革兰氏阴性细菌的基于LPS的疫苗。 许可的亚单位百日咳疫苗提供的保护在个体基础上是不完全的，可能是由于缺乏抗毒素抗体的直接杀菌作用。然而，随着疫苗广泛使用而发生的群体免疫提供了几乎完全的保护。另外一种疫苗成分诱导杀菌抗体，如抗LPS，可以在个体基础上增加疫苗效力。为此B。百日咳和B.支气管败血症LPS的分离和分析。B。百日咳杆菌仅表达由12种糖组成的核心区糖（OS）。B。支气管败血症，更容易培养和具有更好的产量，表达具有与B几乎相同的核心结构的LPS。（1）Fuc 4 NMe的甲基化，B中为100%。在B组百日咳的发病率只有50%。（2）B组Hep的磷酸化程度约为30%。支气管败血症，而在B. Petrussis它不是磷酸化的。B。支气管炎LPS大多进一步被O-特异性链（O-SP）取代。对于该研究，仅使用不含O-SP的游离核级分。两种B的结合物。百日咳OS和B。支气管败血症核心通过使其还原末端Kdo部分与结合至BSA的氨氧基接头反应来制备。两种偶联物平均每个BSA分子掺入10条糖链，均与抗B反应。百日咳和抗BSA血清具有同一性，并且两者在小鼠中具有免疫原性，诱导相似的抗体水平，通过ELISA测量。

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 their non-reducing end saccharide structure; 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 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 into mice as saline solutions, at a fraction of an estimated human dose, induced antibodies to the homologous O-SP. These methodologies can be applied to prepare LPS-based vaccines against other Gram-negative bacteria. The protection afforded by licensed subunit pertussis vaccines is incomplete on an individual basis, likely due to the lack of direct bactericidal effect of anti toxin antibodies. However herd immunity, that occurs with wide vaccine uasage, provides the almost complete protection. An additinal vaccine component inducing bactericidal antibodies such as anti LPS, could increase vaccine efficacy on an individual basis. To that end B. pertussis and B. bronchiseptica LPS were isolated and analyzed. B. pertusis expresses only the core region saccharide (OS) composed of 12 sugars. B. bronchiseptica, easier cultured and with better yields, expresses LPS with almost identical core structure to that of B. pertussis, though we observed small variations: (1) the methylation of Fuc4NMe, 100% in B. pertussis is only 50% in B. bronchiseptica; (2) Hep is phosphorylated in about 30% in B. bronchiseptica,, while in B. petrussis it is not phosphorylated. B. bronchispetica LPS is mostly further substituted by O-specific chains (O-SP). For this study only a free core fraction, with no O-SP was used. Conjugates of both B. pertussis OS and B. bronchiseptica core were prepared by reacting their reducing-end Kdo moieties with an aminooxy linker bound to BSA. Both conjugates incorporated an average of 10 saccharide chains per BSA molecule, both reacted with anti-B. pertussis and anti-BSA sera with a line of identity and both were immunogenic in mice inducing similar antibody levels, measured by ELISA.