Chattopadhayay R, Kaur S, Ganguly N K, Mahajan R C
Department of Parasitology, Postgraduate Institute of Medical Education & Research, Chandigarh.
Indian J Med Res. 1996 Dec;104:349-54.
The axenic amastigotes of an Indian strain of L. donovani have been generated from the promastigote form at 37 degrees C in RPMI-1640 medium, pH 6.0 +/- 0.5, supplemented with 10 per cent heat inactivated foetal calf serum and these are being maintained through serial subculturing under the same conditions. The present study was carried out to differentiate axenic amastigote from the promastigote stage on the basis of their antigenic constitution and also to look for any immunoreactive antigen(s) specific to axenic amastigotes. SDS-PAGE analysis revealed a few stage specific and some conserved antigenic fractions in both the stages. On immunoblotting with immune sera raised against the membrane fractions of the axenic amastigotes, the 200 kDa antigenic fraction of axenic amastigote was found to be highly reactive. When the immune sera raised against the membranes of both stages were checked by immunofluorescence no cross reactivity was observed at higher dilutions. These findings showed that there are some antigenic diversities as well as similarities between the two stages of L. donovani cultured in vitro. Also, the 200 kD fraction of axenic amastigote appeared to be an immunodominant antigen specific to that stage.
杜氏利什曼原虫印度株的无菌无鞭毛体已在37℃、pH 6.0±0.5的RPMI - 1640培养基中由前鞭毛体形式产生，该培养基补充有10%热灭活胎牛血清，并且通过在相同条件下连续传代培养来维持。本研究旨在根据其抗原构成区分无菌无鞭毛体和前鞭毛体阶段，并寻找任何特定于无菌无鞭毛体的免疫反应性抗原。SDS - PAGE分析显示两个阶段都有一些阶段特异性和一些保守的抗原组分。在用针对无菌无鞭毛体膜组分产生的免疫血清进行免疫印迹时，发现无菌无鞭毛体的200 kDa抗原组分具有高度反应性。当通过免疫荧光检查针对两个阶段的膜产生的免疫血清时，在较高稀释度下未观察到交叉反应。这些发现表明，体外培养的杜氏利什曼原虫的两个阶段之间存在一些抗原差异以及相似性。此外，无菌无鞭毛体的200 kD组分似乎是该阶段特有的免疫显性抗原。
