日本血吸虫尾蚴体外培养细胞的形态学与抗原性初步研究

目的探索日本血吸虫尾蚴细胞培养技术，分析培养细胞的形态特征与抗原性。方法采用改良RPMI-1640培养基对日本血吸虫尾蚴细胞作体外培养，观察培养细胞的生长特性和一般形态，取培养3周的细胞作HE染色和超微结构观察，并用SDS—PAGE、Western blot技术、免疫荧光和凝集试验检测其抗原性。结果在培养早期，细胞呈半悬浮状并向培养瓶中心聚集生长，观察到细胞分裂增殖现象，随着’培养时间延长，细胞生长繁殖速度渐渐减慢，培养至8周，多数细胞死亡。培养3周的细胞，大小不均，呈多形性，大部分细胞呈高核质比率，仅少数细胞胞浆较丰富。电镜观察培养细胞表面呈现特殊结构。SDS—PAGE和Immuno blot分析表明：尾蚴细胞与尾蚴虫体的蛋白带谱大体相似，感染兔血清对尾蚴虫体和尾蚴培养细胞抗原分子识别的条带略有不同。用尾蚴培养细胞与感染兔血清作凝集试验和免疫荧光检测，二者均可出现特异性结合。结论日本血吸虫尾蚴细胞在体外出现有限增殖，该细胞在血吸虫病的免疫诊断中具有潜在的应用价值。

Preliminary Study on Morphology and Antigenicity of Cultured Cells from Schistosoma Japanicum Cercaria in Vitro

Objective To optimize the conditions of cell culture for cercariae of Schistosome japonicum,and to identify its morphological characteristics and antigenicity.Methods Cells were cultured in modified RPMI-1640 medium.The growing feature and general morph were observed.After 3 weeks,cercariae cells were observed with HE staining and ultramicrostructure inspection.Cells antigenicity was detected by immunofluorescence and western-blot.Results In the early stage of culture,cells were growing semi-suspendingly with assembling towards center of flask and obvious division and proliferation of cells was observed.Cell growth speed decreased and the majority of cells died or degraded when cells were cultured until eight weeks.Morphous and sizes of cercariae cells were not uniform and showed multi-morphous after three weeks culture.Most cells had a high nuclear-to-cytoplasm ratio with few cells possessing rich cytoplasm.Under the scanning electron microscopy,some cells surface took on special structure.Cells and worm bodies of cercariae had similar protein bands and were identified differently using infected rabbit serum by Western-blot analysis.Infected rabbit sera bonded to cercariae cultured cells specially by agglutination trial and immunofluorescence assay.Conclusion Cercariae cells of Schistosome japonicum show proliferation trend,which display diagnosis potential in schistomasis.