Salem S A, el-Kowrany S I, Ismail H I, el-Sheikh T F
Department of Parasitology, Faculty of Medicine, Tanta University, Tanta, Egypt.
J Egypt Soc Parasitol. 2001 Apr;31(1):133-44.
The expression of heat shock proteins (HSPs) and their immunogenic role in host parasite relationship has been studied in T. spiralis infection in mice. Two groups of infective stage larvae were incubated at 37 degrees C or at 43 degrees C for 3 hours in a nutrient medium. Crude somatic extracts and excretory/secretory (E/S) products were obtained. Quantitative estimation and in vitro translation of mRNA were carried out. Crude somatic extracts, E/S products and in vitro translated proteins were all subjected to SDS-PAGE and immunoelectrophoresis against a monoclonal antibody to HSP70. They were further used to immunize mice which were then infected with T. spiralis. The degree of immunization was assessed by counting adult worms, muscle larvae and estimating total gamma globulins of mice. SDS-PAGE revealed intense peaks of 71 KDa and 81 KDa in all proteins obtained from heat shocked larvae. These proteins have been proved to involve HSP70 as manifested by their strong reactivity with anti-HSP70. In vitro translated products of heat shocked larvae (at 43 degrees C) proved to be strongly immunogenic as evidenced by the lower adult worm, muscle larval counts and higher total gamma globulins in sera of mice in comparison with the control non immunized group as well as to other larval antigens incubated at 37 degrees C. So, exposure to heat, can induce the synthesis of heat shock proteins which can defend the organism against environmental stress. Meanwhile, these heat shock proteins can render the host more refractory to reinfection.
热休克蛋白(HSPs)在小鼠旋毛虫感染中的表达及其在宿主与寄生虫关系中的免疫原性作用已得到研究。将两组感染期幼虫在营养培养基中于37℃或43℃孵育3小时。获得粗制体细胞提取物和排泄/分泌(E/S)产物。进行mRNA的定量估计和体外翻译。粗制体细胞提取物、E/S产物和体外翻译的蛋白质均进行SDS-PAGE,并针对抗HSP70单克隆抗体进行免疫电泳。它们进一步用于免疫小鼠,然后用旋毛虫感染这些小鼠。通过计数成虫、肌幼虫并估计小鼠的总γ球蛋白来评估免疫程度。SDS-PAGE显示,从热休克幼虫获得的所有蛋白质中均有71 kDa和81 kDa的强峰。这些蛋白质已被证明与HSP70有关,因为它们与抗HSP70具有强烈反应性。热休克幼虫(43℃)的体外翻译产物被证明具有很强的免疫原性,与未免疫的对照组以及在37℃孵育的其他幼虫抗原相比,小鼠血清中的成虫、肌幼虫数量较低,总γ球蛋白较高即可证明。因此,暴露于热可以诱导热休克蛋白的合成,这些蛋白可以保护生物体免受环境压力。同时,这些热休克蛋白可以使宿主对再次感染更具抵抗力。