Roberts M, Bacon A, Rappuoli R, Pizza M, Cropley I, Douce G, Dougan G, Marinaro M, McGhee J, Chatfield S
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom.
Infect Immun. 1995 Jun;63(6):2100-8. doi: 10.1128/iai.63.6.2100-2108.1995.
We examined the capacity of a genetically detoxified derivative of pertussis toxin (PTX), PT-9K/129G, to act as a mucosal adjuvant for an intranasally (i.n.) administered tetanus vaccine. Groups of mice were immunized i.n. with the nontoxic C-terminal 50-kDa portion of tetanus toxin (fragment C [Frg C]) either alone or mixed with PT-9K/129G, PTX, or cholera toxin (CT) or were immunized subcutaneously (s.c.) with an equivalent amount of Frg C adsorbed to alhydrogel. In response to a single immunization, mice receiving Frg C plus PT-9K/129G or CT i.n. and parenterally immunized mice developed high-titer (> 20,000) anti-Frg C antibodies, whereas mice immunized i.n. with Frg C plus PTX or with Frg C alone seroconverted only after being boosted. The serum anti-Frg C response was dominated by immunoglobulin G1 (IgG1) in mice immunized with Frg C plus PT-9K/129G, with Frg C plus PTX, or s.c. In contrast, IgG1, IgG2a, and IgG2b contributed almost equally to the Frg C response when CT was the adjuvant. Anti-Frg C IgE was detected only in the sera of mice immunized i.n. with Frg C plus PTX and immunized s.c. with Frg C plus alhydrogel. High levels of IgA antibodies were present in nasal lavage fluid from mice immunized i.n. with Frg C plus PT-9K/129G, PTX, or CT but not in that from mice given Frg C alone i.n. or parenterally. The mucosal adjuvanticity of PT-9K/129G was manifested in inbred as well as outbred mice. A single i.n. dose of Frg C plus either PT-9K/129G or PTX (with high specific activity) was sufficient to protect all immunized mice from tetanus toxin challenge, in contrast to the case for mice that received Frg C alone i.n. We conclude that the pertussis toxin analog PT-9K/129G, which is devoid of ADP-ribosyltransferase activity, is a potent mucosal adjuvant for vaccines delivered via the respiratory tract.
我们检测了百日咳毒素（PTX）的一种基因解毒衍生物PT-9K/129G作为鼻内（i.n.）接种破伤风疫苗的黏膜佐剂的能力。将小鼠分组，分别经鼻内接种破伤风毒素无毒C末端50 kDa片段（片段C [Frg C]）单独制剂，或与PT-9K/129G、PTX或霍乱毒素（CT）混合的制剂，或者经皮下（s.c.）接种等量吸附于氢氧化铝凝胶的Frg C。单次免疫后，经鼻内接种Frg C加PT-9K/129G或CT的小鼠以及经胃肠外免疫的小鼠产生了高滴度（> 20,000）的抗Frg C抗体，而经鼻内接种Frg C加PTX或单独接种Frg C的小鼠仅在加强免疫后才出现血清转化。在用Frg C加PT-9K/129G、Frg C加PTX免疫的小鼠或经皮下免疫的小鼠中，血清抗Frg C反应以免疫球蛋白G1（IgG1）为主。相比之下，当CT作为佐剂时，IgG1、IgG2a和IgG2b对Frg C反应的贡献几乎相等。仅在经鼻内接种Frg C加PTX以及经皮下接种Frg C加氢氧化铝凝胶免疫的小鼠血清中检测到抗Frg C IgE。在用Frg C加PT-9K/129G、PTX或CT经鼻内免疫的小鼠的鼻腔灌洗液中存在高水平的IgA抗体，而在经鼻内或胃肠外单独给予Frg C的小鼠的鼻腔灌洗液中则没有。PT-9K/129G的黏膜佐剂活性在近交系和远交系小鼠中均有体现。与经鼻内单独接种Frg C的小鼠不同，经鼻内单次给予Frg C加PT-9K/129G或PTX（具有高比活性）足以保护所有免疫小鼠免受破伤风毒素攻击。我们得出结论，缺乏ADP-核糖基转移酶活性的百日咳毒素类似物PT-9K/129G是通过呼吸道递送的疫苗的有效黏膜佐剂。
