SjAWMAg免疫血清和吡喹酮联用体内杀不同发育期血吸虫的实验研究

EXPERIMENTAL STUDY ON SYNERGIC EFFECT OF IMMUNE SERUM AGAINST SURFACE MEMBRANE ANTIGEN OF SCHISTOSOMA JAPONICUM ADULT AND PRAZIQUANTEL IN KILLING DIFFERENT DEVELOPMENTAL STAGE WORMS

目的 观察日本血吸虫成虫表膜抗原(SjAWMAg)免疫血清与吡喹酮合用对不同发育期血吸虫的杀虫效果,探讨体液免疫在吡喹酮杀虫中的作用。 方法 提取 SjAWMAg,并用 SDS PAGE 对其进行鉴定,制备高滴度(1∶25 600)SjAWMAg抗血清,采用Western blot分析该血清与肺期童虫、肝期童虫及成虫抗原的反应性;将 SjAWMAg抗血清与吡喹酮合用治疗血吸虫感染后第2 d、第14 d和第35 d的小鼠,在感染后第49 d剖杀小鼠收获虫体,计算减虫率和减卵率。结果 Western blot分析表明,SjAWMAg抗血清与肺期及肝期童虫抗原存在交叉反应; 联合用药的 3 个治疗组(感染2、14、35 d)与单用吡喹酮组比较杀虫作用均有显著提高,减虫率分别提高了 38.79%、29.80%和 46.82%,其中以感染35 d治疗组提高最显著;减卵率分别提高了 62.04%,42.78%和 29.81%,以感染 2 d治疗组更为显著。结论抗SjAWMAg多克隆抗体与吡喹酮联合使用,均可提高吡喹酮对不同发育期血吸虫的杀灭效果,揭示体液免疫在吡喹酮杀血吸虫过程中具有重要作用,同时提示这种联合用药可能解决单用吡喹酮治疗血吸虫早期感染效果不佳的难题。

Objective To observe the synergic effect of the antibodies against surface membrance antigen of Schistosoma japonicum (Sj) adult worm and praziquantel(PZQ) in killing different developmental stage worms and to study the role of humoral immunity in the schistosomicidal process of praziquantel. Methods Surface membrance antigen of Sj adult were extracted and characterized by SDS PAGE.Serum from rabbit immunized with surface membrance antigen of Sj adult was prepared. The cross reactions betwwen the immune serum and different developmental stage antigens of Sj were analyzed by Western blot. The groups of experimental mice were treated with PZQ plus immune serum injection at 2 d, 14 d and 35 d after infection, respectively. All mice were sacrificed 49 d after infection. Worm burden and liver eggs per gram (LEPG) were counted. Results Worm burden and egg burden of the experimental mice treated at 2 d, 14 d and 35 d by combining usage of immune serum and PZQ were dramatically reduced, compared with those only treated by praziquantel. The immune serum increased the schistosomicidal activity of PZQ against different developmental stage worms (2 d,14 d and 35 d) by 38.79%, 29.80% and 46.82%, respectively .Correspondingly, egg burden in liver decreased in each group by 62.04%, 42.78% and 29.81%. Conclusion Immune serum against surface membrance antigen of Sj adult combined with praziquantel can improve the schistosomicidal effect. That indicated that the humoral immunity play a key role in the schistosomicidal process of PZQ.