Ohta Nobuo, Kumagai Takashi, Maruyama Haruhiko, Yoshida Ayako, He Yongkan, Zhang Renli
Department of Molecular Parasitology, Nagoya City University Graduate School of Medical Sciences, Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601, Japan.
Parasitol Int. 2004 Jun;53(2):175-81. doi: 10.1016/j.parint.2004.01.007.
Vaccine development by the use of calpain of Schistosoma japonicum has been tried in our laboratory. We cloned cDNA encoding the heavy chain of S. japonicum calpain, and prepared recombinant molecule of a possible vaccine region of the heavy chain. When BALB/c mice were immunized with our recombinant calpain of S. japonicum with Freund's complete adjuvant, we observed significant reduction in worm burden (41.2% reduction, P<0.05), and also significant anti-fecundity effects. In this sense, calpain of S. japonicum seems to have infection control as well as anti-disease effects. Mechanisms of vaccine effects of calpain remain to be clarified, however, several effector mechanisms are suspected. In immunized mice, raised level of iNos expression was observed, while adhesion of peritoneal exudates cells were also observed in the presence of calpain-immunized sera, suggesting the possibilities of both cellular and humoral protective mechanisms. We examined tissue distribution of calpain in various developmental stages of S. japonicum. Strong signal was observed around excretory grand of cercariae, and they secreted calpain during their migratory movement tested in vitro. Together with the findings, calpain seems to induce larvicidal effects in the immunized mice. We observed time-course kinetics of antibody production against vaccine candidates in experimental S. japonicum infection in pigs. Although significant levels of antibody production were observed for paramyosin and GST, no significant antibody production was observed for calpain. This suggests that calpain is less immunogenic, and route of immunization and/or choice of adjuvant are important in future trials of calpain vaccine.
我们实验室已尝试利用日本血吸虫钙蛋白酶进行疫苗研发。我们克隆了编码日本血吸虫钙蛋白酶重链的cDNA,并制备了重链可能的疫苗区域的重组分子。当用弗氏完全佐剂将我们制备的日本血吸虫重组钙蛋白酶免疫BALB/c小鼠时,我们观察到虫体负荷显著降低(降低了41.2%,P<0.05),并且还有显著的抗生育力作用。从这个意义上讲,日本血吸虫钙蛋白酶似乎具有感染控制以及抗病作用。然而,钙蛋白酶疫苗效应的机制仍有待阐明,不过怀疑有几种效应机制。在免疫小鼠中,观察到诱导型一氧化氮合酶(iNos)表达水平升高,同时在存在钙蛋白酶免疫血清的情况下也观察到腹膜渗出细胞的黏附,这表明细胞和体液保护机制都有可能。我们研究了钙蛋白酶在日本血吸虫各个发育阶段的组织分布。在尾蚴排泄孔周围观察到强信号,并且它们在体外迁移运动过程中分泌钙蛋白酶。综合这些发现,钙蛋白酶似乎在免疫小鼠中诱导杀幼虫作用。我们观察了猪实验性日本血吸虫感染中针对候选疫苗抗体产生的时间进程动力学。虽然观察到副肌球蛋白和谷胱甘肽S-转移酶(GST)有显著水平的抗体产生,但未观察到钙蛋白酶有显著的抗体产生。这表明钙蛋白酶的免疫原性较低,并且免疫途径和/或佐剂的选择在未来钙蛋白酶疫苗试验中很重要。