基因重组丝虫融合蛋白抗原抗丝虫的保护性免疫及用于丝虫病血清学诊断、监测的研究

EVALUATION OF RECOMBINANT CHITINASE ANTIGEN ANTIFILARIAL IMNUNE PROTECTIVE AND AS SEROLOGICAL DIAGNOSIS,SURVEILLANCE OF LYMPHATIC FILARIASIS

目的:测定基因重组丝虫壳质酶融合蛋白抗原对动物感染丝虫的保护性免疫作用和用于丝虫病血清学诊断和监测的评价。方法:用基因重组壳质酶抗原免疫沙鼠,ELISA方法检测动物及人血清中的抗体水平,SDS-聚丙烯酰胺凝胶电泳和免疫印迹试验测定特异的蛋白质分子。结果:壳质酶抗原可诱导动物体内产生特异的抗丝虫抗体,在用丝虫感染期幼虫攻击感染后,这种特异的抗体可使沙鼠血循环中微丝蚴(Mf)出现的时期明显推迟,MI数显著减少,成虫数也减少。壳质酶抗原与微丝蚴可溶性抗原(Mf-xt)检测Mf阳性沙鼠、正常沙鼠、Mf血症患者、流行区非Mf血症者和非流行区正常人的血清抗体水平中壳质酶检测的结果分别为100％,0,100％,5％和O,则Mf-xt为80％,0,80％,20％和5％。证明壳质酶抗原较Mf-xt敏感性高,特异性强。结论:壳质酶融合蛋白抗原免疫沙鼠具部分的抗丝虫感染的保护性免疫作用。壳质酶抗原同样可用于丝虫病的血清学诊断和监测。

Objective: To determine wheher immunization with Rec- chitinase could protect jirds against filarial resulting from infection with B. malayi, evaluation of chitinase antigen using serologicaldiagnosis and surveillance of lymphatin filariasis.Methods: Thejirds were immunized with chitinase, antibodies level of human and jirds sera were determined br ELISA with chitimase and Mf soluble extract antigens(Mf- xt). Special protein molecular weight was-determined with SDS- PAGE, western bolt.Results: Specific antibody can be induced in jirds by immunization with chitinase, while immunization with chitinase elicited at least partial protection antinst microfilaremia and delayed the onset and readuced the level of Mf resulting in circulation from infection with the B.malayi and also reduced burdens of adult worm.The comparison usefulness of achitinase with Mf - xt for the serologic diagnosis and surveillance of lymphatic filariasis, the result shown that chitinase determinde Mf positive jirds(100%) contained specific antibody to chitinase, normal jirds can test nothing antibody, microfilaremia was 100%, amicrofilaremia donors living in endemic area was 5% and health students living in umendemic area was 0, however Mf - st detected 80%, 0, 80%, 20% and5% respectively.The result identificated thatchitinase more sensitive and special than Mf - xt to determine filariae antibodies. Conclusion: Our results indicate that recombinant fusion protein filarial antigen was partially protective and is suitable for detection of active occult or patent lymphatic filariasis with daytime blood samples.