Flanigan T P, King C H, Lett R R, Nanduri J, Mahmoud A A
Department of Medicine, Case Western Reserve University, Cleveland, Ohio.
J Clin Invest. 1989 Mar;83(3):1010-4. doi: 10.1172/JCI113942.
Freeze-thaw (FT)-disrupted schistosomula or their membrane extract induced significant resistance in mice to Schistosoma mansoni infection (34 and 25%, respectively) without the use of adjuvant. Antigens identified in schistosome extracts by sera from immunized animals were then evaluated for protective potential. Immunization with schistosomal antigens of 97 and 68-70 kD resulted in significant protection that was equivalent to that obtained by FT schistosomula. Since the 97-kD antigen was suggested to be parasite paramyosin, we used a biochemical technique to purify this muscle protein. Purified schistosome paramyosin ran as a single band on 10% SDS-PAGE and was recognized both by sera from mice immunized with FT schistosomula and a polyclonal antiserum raised against the 97-kD parasite protein. Preincubation of schistosome paramyosin with sera from mice immunized with FT schistosomula resulted in the removal of reactivity with the 97-kD protein in crude worm extracts. Paramyosin was identified by Western blotting to be in the tegument of schistosomula. The purified schistosome paramyosin resulted in significant protection in three separate experiments (24, 46, and 53%) without the use of adjuvant. Addition of BCG to paramyosin resulted in enhanced protection.
冻融(FT)破坏的童虫或其膜提取物在不使用佐剂的情况下,能使小鼠对曼氏血吸虫感染产生显著抗性(分别为34%和25%)。然后,对免疫动物血清在血吸虫提取物中鉴定出的抗原的保护潜力进行了评估。用97 kD和68 - 70 kD的血吸虫抗原进行免疫可产生显著保护作用,其效果与FT童虫相当。由于97 kD抗原被认为是寄生虫副肌球蛋白,我们采用生化技术纯化了这种肌肉蛋白。纯化的血吸虫副肌球蛋白在10% SDS - PAGE上呈现为单一条带,并且能被用FT童虫免疫的小鼠血清以及针对97 kD寄生虫蛋白产生的多克隆抗血清识别。用FT童虫免疫的小鼠血清与血吸虫副肌球蛋白预孵育后,可消除与粗制虫体提取物中97 kD蛋白的反应性。通过蛋白质印迹法鉴定出副肌球蛋白存在于童虫的体表。纯化的血吸虫副肌球蛋白在三个独立实验中均产生了显著保护作用(分别为24%、46%和53%),且无需使用佐剂。向副肌球蛋白中添加卡介苗可增强保护作用。