Immunodiagnostic value and properties of an antigen isolated with affinity chromatography using a monoclonal antibody to Trichinella spiralis

制备了分离旋毛虫特异性抗原的单克隆抗体。根据其反应性和特异性选择IgG_1类单克隆抗体TS32D12。采用TS32D12亲和层析技术，从t -s肌幼虫(Ts-L_1) α -坚韧细胞中分离出160kDa分子（Ts-alpha 160），并对13例患者感染后33-109 d的血清样本进行免疫诊断。在所有病例中检测到的Ts-alpha 160抗体滴度均高于ts -粗抗原抗体滴度。将Ts-alpha 160应用于ELISA，可消除确诊蛔虫患者血清中ts -粗抗原的假阳性反应。另一方面，在感染后第14天的成虫、胚胎和肌肉幼虫中均未检测到160kDa分子。然而，在肌肉晚期的α -硬结体中证实了这一点，至少从第21天到第180天有更多的vae。感染t - alpha 160的BALB/c小鼠在第4周出现低滴度抗体反应，随后在第8周滴度迅速增加。这些结果表明，Ts-alpha 160可作为感染后5 ~ 6周人旋毛虫病的独特抗原进行免疫诊断。此外，发现Ts-alpha 160是由Ts-L_1衍生的糖蛋白和E-S产物组成，TS32D12在100℃，2 min处理后似乎失去了识别的表位。电镜下免疫金法鉴定了Ts-alpha 160蛋白，该蛋白位于ts - l1的棘细胞α颗粒中。在被动转移TS32D12和主动转移Ts-alpha 160的实验组中，蠕虫的恢复没有显著降低。将新鲜分离的弓形虫L_1在含栓塞铵的PBS中孵育，大量沉淀积累并粘附在弓形虫的口腔末端，通过免疫荧光染色观察其对TS32D12的反应。少

Monoclonal antibodies were generated for the isolation of specific antigen from Trichinella spiralis(T.s). A monoclonal antibody (TS32D12) of IgG_1 class was selected according to its reactivity and specificity. We isolated 160kDa molecule (Ts-alpha 160) from alpha -stichocyte of T.s muscle larvae(Ts-L_1) with affinity chromatography using TS32D12 and examined immunodiagnostic value of Ts-alpha 160 against serum samples which had been obtained from 13 patients 33-109 days after infection. Antibodies against Ts-alpha 160 were detected with higher titers than those against Ts-crude antigen in every case. The application of Ts-alpha 160 to the ELISA eliminated false-positive reactions which were observed against Ts-crude antigen in serum samples from patients with proven helminthiasis. On the other hand, the 160kDa molecule was not detected in adult worms, embryos, nor in muscle larvae at day 14 post infection. However, it was demonstrated in alpha -stichosome of the late muscle stage lar … More vae from day 21 at least up to day 180. Antibody responses of infected BALB/c mice of T.s against the Ts- alpha 160 became detectable with low titers at week 4, followed by a rapid increased in titers at week 8. These results suggested that Ts-alpha 160 can be utilized as a unique antigen for the immunodiagnosis of human trichinosis from about 5-6 week after infection. In addition, Ts-alpha 160 was found consist of glycoprotein and E-S product derived from Ts-L_1, the epitopes recognized by the TS32D12 seemed to be lost after treatment at 100ﾟC, 2 min. The Ts-alpha 160 was elucidated by the immunogold method using electron microscope that is located in alpha granules of stichocyte of Ts-L_1. There were no significant decrease in recovery of worms among experimental groups of mice both passively transferred TS32D12 and actively transferred Ts-alpha 160. Freshly isolated T.s L_1 were incubated containing ammonium embelate in PBS, large precipitates accumulated and adhered to the oral end of the worms,and were observed to react to TS32D12 by immuno-fluorescence stain. Less