旋毛虫肌肉期幼虫分泌排泄物中特异性抗原的分析

Analysis of Specific Antigens Extracted from Excretory Secretory Products of Trichinella spiralis Muscle Larvae

用胃蛋白酶消化旋毛虫感染鼠肌肉，获得肌肉期幼虫。３７℃，用ＲＰＭＩ１６４０培养基培养幼虫，控制虫体死亡率低于５％。每２４ｈｒ收集上清培养液，即为分泌排泄抗原（ＥＳＡ）。共获１０天次ＥＳＡ（ＥＳＡ＿（１ｄ～１０ｄ））。应用ＳＤＳ－ＰＡＧＥ及氨银染色技术进行蛋白组份分析显示：各天次ＥＳＡ的主要蛋白区带数及位置基本相同，分子量范围在９６～１４ｋｄ，主带３条，分别为４８，５３和５８ｋＤ蛋白组份。应用Ｗｅｓｔｅｒｎ－ｂｌｏｔ技术进行免疫识别分析显示：旋毛虫病人血清对各天次ＥＳＡ的免疫识别结果相似，识别的抗原组份有７～８条区带，均有上述３条主带。正常人、蛔虫病人和囊虫病人血清均未能识别任何区带。肺吸虫病人血清能识别２～３条抗原区带，但其分子量位于２４～１４ｋＤ之间。血吸虫病人血清也能识别２～３条抗原区带，其分子量位于１８～１４ｋＤ之间。结果表明：在严格控制死亡虫体抗原污染条件下，较长期内培养肌肉期幼虫获得的各天次ＥＳＡ，其成份和免疫识别效果基本无改变，各天次ＥＳＡ中的４６～５８ｋＤ蛋白均具有良好的特异性。

Muscle larvae were obtained from infected mice by peptic digestion. The larvae were culturedin RPMI1640 medium at 37℃. The medium was changed every 24 hours for 10 days in order to remove thedead larvae Trichinella spiralis,and the rate of dead larvae must be no more than 5%。The collected super-natant,excretory-secretory antigen:ESA1d,ESA26,ESA46,ESA6d,ESA8d and ESA10d were analysed bySDS-PAGE with silver staining,Results showed that the stained bands of these kinds of ESA were similar innumber and site. The molecular weight of these banks were between 14～96 kD with three chief bands in 48,53 and 58 kD.ESA1d,ESA2d,ESA5d,ESA8d and ESA10d were analysed by Western-blot and the results showedthat the effect of immune recognition were similar.The sera from patients with trichinosis were recognizedwith 7～8 antigenic bands in chief bands of 46,48,50/53 and 58 kD. The sera from health persons and pa-tients with ascariasis or cysticerosis showed no any bands The sera from patients with paragonimiasis wererecognized with 2～3 antigenic bands at 14～24 kD,and those from patients with schistosomiasis with 2～3antigenic bands at 14～18 kD. These results indicate that the proteins of ESA between 46～58 kD were highlyspecific antigens.