Douce G, Giuliani M M, Giannelli V, Pizza M G, Rappuoli R, Dougan G
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, UK.
Vaccine. 1998 Jul;16(11-12):1065-73. doi: 10.1016/s0264-410x(98)80100-x.
Using a fixed dose of antigen, the immune response to detoxified mutants of LT-WT following intranasal (i.n.), subcutaneous (s.c.) and oral (i.g.) immunisation has been studied. When given i.n., both LT-WT and mutant toxin, K63, generated significant levels of toxin-specific IgG in the serum, and the levels of IgA in nasal and lung lavages were greater than those induced by rLT-B. In comparison, i.g. immunisation of mice with a similar quantity of either LT-WT or K63 toxin induced barely detectable levels of IgG in the sera. However, if the amount of protein used for i.g. immunisation was increased tenfold, relatively good levels of toxin-specific IgG were induced in the sera by both LT-WT or K63. Low levels of toxin-specific IgA were also observed in intestinal washes from these mice. Western blotting of the sera, using the native toxin as an antigen, demonstrated the presence of both anti-A and anti-B subunit antibodies. Most significantly, toxin-neutralising antibodies were induced in the serum, with the strongest activity being induced by the LT-WT, an intermediate activity induced by mutant K63 and a lower response by rLT-B. Together, these data show that ADP-ribosyltransferase is not necessary for mucosal immunogenicity of these proteins, and that the i.n. route of immunisation is more effective than the i.g. route of immunisation for the generation of both systemic (IgG) and mucosal (IgA) immune responses.
使用固定剂量的抗原，研究了鼻内（i.n.）、皮下（s.c.）和口服（i.g.）免疫后对LT-WT解毒突变体的免疫反应。鼻内给药时，LT-WT和突变毒素K63均在血清中产生了显著水平的毒素特异性IgG，并且鼻洗液和肺灌洗液中的IgA水平高于rLT-B诱导的水平。相比之下，用相似量的LT-WT或K63毒素对小鼠进行口服免疫，血清中诱导出的IgG水平几乎检测不到。然而，如果用于口服免疫的蛋白量增加10倍，LT-WT和K63在血清中均诱导出相对较好水平的毒素特异性IgG。在这些小鼠的肠道冲洗液中也观察到低水平的毒素特异性IgA。用天然毒素作为抗原对血清进行蛋白质印迹分析，结果显示存在抗A和抗B亚基抗体。最重要的是，血清中诱导出了毒素中和抗体，其中LT-WT诱导的活性最强，突变体K63诱导出中等活性，rLT-B诱导的反应较低。总之，这些数据表明ADP-核糖基转移酶对于这些蛋白质的黏膜免疫原性不是必需的，并且鼻内免疫途径在产生全身（IgG）和黏膜（IgA）免疫反应方面比口服免疫途径更有效。
