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目的:研究大理猪源旋毛虫肌幼虫(Trichinela spiralis musclelarvae)3种抗原:虫体可溶性抗原(body soluble anti-gen,BSA)、表面抗原(surface antigen,SA)和排泄分泌抗原(excretory-secretory antigen,ESA)的蛋白组分及其之间的差异,并进一步比较分析其免疫反应性,为分离和筛选出免疫原性和免疫反应性强的旋毛虫肌幼虫抗原成分打基础。方法:采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),对肌幼虫3种抗原蛋白成分进行分析;采用蛋白印迹(Western-blot)对成虫3种抗原蛋白组分进行免疫反应性分析。结果:SDS-PAGE电泳结果:BSA显示30条蛋白染色条带,分子量范围在100 kDa～12 kDa之间,其中主带11条;SA显示10条主要蛋白染色条带,分子量范围在105 kDa～28 kDa之间;ESA显示20条蛋白染色条带,分子量范围在112 kDa～10 kDa之间,其中主带11条。Western-blot结果:BSA出现25条反应条带,其中分子量为75 kDa、66 kDa、63 kDa、55 kDa、49 kDa、45 kDa、37 kDa、28 kDa、20 kDa、16 kDa的条带着色较深;SA出现7条反应条带;ESA显示14条反应带,其中53 kDa、49 kDa、45 kDa、43 kDa、35 kDa、27 kDa、18 kDa、12 kDa显色明显。结论:旋毛虫肌幼虫BSA与ESA较SA蛋白组分复杂,三者主带部分属低分子量区;免疫反应性的强度SA和ESA高于BSA,说明前两种抗原是旋毛虫肌幼虫刺激机体产生免疫应答的主要靶抗原。
Objective: To study the expression of three antigens of Trichinella spiralis musclelarvae (body soluble anti-gen (BSA), surface antigen (SA) and excretory-secretory antigen , ESA) protein components and their differences, and further comparative analysis of their immunological reactivity, for the isolation and screening of immunogenic and immunocompetent Trichinella spiralis larvae antigen components lay the foundation. Methods: SDS-PAGE was used to analyze the protein components of three kinds of antigens in muscle larvae. Western blotting was used to analyze the protein components of the three adult antigens Immunoreactivity analysis. Results: The result of SDS-PAGE showed that BSA showed 30 protein bands with the molecular weight ranged from 100 kDa to 12 kDa, of which 11 were primary bands. SA showed 10 major protein bands with a molecular weight range of 105 kDa ~ 28 kDa; ESA showed 20 protein bands, the molecular weight range between 112 kDa ~ 10 kDa, of which the main band of 11. Western-blot results showed that there were 25 reaction bands in BSA, and the bands of 75 kDa, 66 kDa, 63 kDa, 55 kDa, 49 kDa, 45 kDa, 37 kDa, 28 kDa, 20 kDa and 16 kDa Seven reaction bands were observed in SA. Seventeen reaction bands were observed in ESA, of which 53 kDa, 49 kDa, 45 kDa, 43 kDa, 35 kDa, 27 kDa, 18 kDa and 12 kDa were evident. Conclusion: The components of BSA and ESA of Trichinella spiralis larvae are more complex than that of SA protein, and the main bands of the three strains belong to low molecular weight region. The intensities of immunoreactive SA and ESA are higher than that of BSA, indicating that the first two antigens are the stimulation of Trichinella spiralis larvae The main target antigen that produces an immune response.