猪带绦虫45W-4BX和TSOL18的高效表达及免疫效果研究

RT-PCR扩增45W-4B和TSOL18基因，PCR截去45W-4B基因的N端信号肽和C端疏水氨基酸序列形成45W-4BX。将45W-4BX和TSOL18 PCR产物分别亚克隆人pGEX-4T-1，用IPTG诱导表达，取产物进行SDS-PAGE和Western blot分析。纯化表达产物制成油佐剂疫苗分别免疫家猪，用25000枚猪带绦虫成熟虫卵进行攻击感染，ELISA检测各组的抗体水平，90d后剖检计算各组的减虫率。结果表明，45W-4BX和TSOL18基因在大肠杆菌中分别以可溶性和包涵体形式获得高效表达，并能被囊虫病人血清所识别。重组蛋白免疫猪15d血清抗体即为阳性，30d左右达到峰值。2种重组抗原的减虫率均在88％以上，与囊虫粗抗原免疫效果相当。这为进一步研制基于45W-4BX和TSOL18的猪囊虫重组基因工程疫苗奠定了基础。

Highly Efficacious Expression and Immune Efficacy of Recombinant 45W-4BX and TSOL18 from Taenia solium Oncosphere

Genes 45W-4B and TSOL18 were amplified by RT-PCR. The truncated 45W-4B named 45W-4BX was acquired by PCR. 45W-4BX and TSOL18 were subcloned into pGEX-4T-1, respectively. Soluble 45W-4BX and TSOL18 in the form of inclusive bodies were detected by SDS- PAGE, Western blot and the results indicated that they could be recognized by sera from cysticercosis patients. Purified antigens 45W-4BX, TSOL18 and GST and cysticercus cellulosae coarse extract were used to immunize pigs, which were experimentally challenged with 25 000 eggs of Taenia solium 28 days postimmunization. Data suggested that antibody level would start to raise at 15 days and reach peak at 45 days postimmunization. Reduction in the number of cysticerci in the groups immunized with recombinant proteins were above 88 %. Level of protection of groups vaccinated with TSOL18 and 45W-4BX did not show significant difference in comparison with the cysticercus cellulosae coarse extract. Recombinant antigens TSOL18 and 45W-4BX were primarily elucidated and their protective efficacy was confirmed.