伯氏疟原虫红外期体外培养的研究

IN VITRO CULTIVATION OF THE EXOERYTHROCYTIC STAGE OF PLASMODIUM BERGHEI

本文报告在国内首次建成了胚肺细胞-伯氏疟原虫红外期体外培养系统.用胰酶消化法自人工流产胎儿分离出胚肺细胞后,建立Elu 8801细胞株。斯氏按蚊叮咬伯氏疟原虫ANKA株感染的昆明小鼠18～21d后,在无菌条件下解剖出唾腺,制备子孢子悬液,接种于单层培养的胚肺细胞。培养48h后,可见红外期裂殖体。72h后,部分裂殖体内已形成成熟的裂殖子,将培养上清经腹腔接种健康小鼠,可使之感染疟疾并可经血传感染其它小鼠。用第2代感染小鼠血饲喂按蚊,蚊体内可产生子孢子。表明本室建立的人胚肺细胞林对P.b.ANKA株易感染,并能支持其红外期发育成熟,产生有感染力的红外期裂殖子。

This paper reports on an in vitro culture system for the exoerythrocytic (EE) stage of Plasmodium berghei (P.b.) using embryonic lung cells. The system was first developed by our laboratory in China. The embryonic lung cells were isolated by trypsin digestion of a human embryonic lung obtained from a therapeutic abortion case and was designated as cell line Elu 8801. Anopheles stephensi mosquitoes were infected by biting P. b. ANKA strain infected Kunming mice and after 18-21 days were dissected under aseptic conditions for preparation of a sporozoite suspension. This suspension was used to inoculate the monolayer cultures of Elu 8801. Regular examination found that following a cultivation for 48 hours, up to 100 multinuclear EE schizonts of P.b. could be observed on 1×1cm cover slide. Seventy-two hours later mature merozoites were seen among part of the schizonts. An intrape-ritoneal inoculation of the supernatant culture medium to mice could induce malaria infection which could be transferred to other mice by blood inoculation. When the mice infected with the second generation were allowed to feed A. stephensi, sporozoites developed in the mosquitoes. The results demonstrate that the human embryonic lung cell line Elu 8801 established in our laboratory is susceptible to P.b. ANKA sporozoites and can support the developmental maturation of EE stages, producing potentially infectious merozoites.