体外培养杜氏利什曼原虫无鞭毛期和期特异性抗原的测定

THE CULTURE OF LEISHMANIA DONOVANI AMASTIGOTE IN VITRO

利什曼原虫无鞭毛期主要寄生在人体内巨噬细胞和网状内皮系统的其它吞噬细胞内。通常,媒介体内的前鞭毛体可经体外培养获得,而对无鞭毛期原虫的体外培养研究自1979年以来曾有不少学者用kuffer′s细胞、皮肤粘膜细胞、骨髓细胞及人外周血液等方法,均只能在较短时间获得成功。国内对无鞭毛期的体外培养,迄今尚无报道。为此,我们对其培养方法进行了研究,并以获得的材料,对杜氏利什曼原虫无鞭毛期的相关抗原作了观察。

We have reported a culture system,in which a BALB/c mouse macrophage cell line is used for continuous propagation of Leishmania donovani amastigotes and a method for their isolation from the cultured material. The macrophage suspension was prepared at a cell density of 10~6 per milliliter, then added to promastigote sediment after centrifugation at a parasite—macrophage ratio of 10:1 and mixed thoroughly. The mixture was incubated at 35℃. The intracellular parasites from this infected cultures can be isolated with high recovery rate. This in vitro culture system has been used for large amount of Leishmania donovani propagation. Then we may use monoclonal antibodies for identification of amastigote stage specific antigen by IFA and Dot —ELISA.