Long G W, Leath S, Schuman R, Hollingdale M R, Ballou W R, Sim B K, Hoffman S L
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland 20814.
In Vitro Cell Dev Biol. 1989 Sep;25(9):857-62. doi: 10.1007/BF02623670.
Plasmodium berghei exoerythrocytic (EE) stages have been cultured in vitro in human continuous cell lines and primary cultures of both human and rat hepatocytes. Although the predominant experimental model of irradiated sporozoite-induced protective immunity is the mouse, P. berghei has not been cultivated in primary mouse hepatocytes or in continuous mouse lines. Because of this, target cells are not available for determining if these immunized mice produce cytotoxic T lymphocytes (CTLs) that recognize P. berghei antigens expressed on hepatocytes in the context of class I major histocompatability (MHC) antigens. We report the development of methods for cultivating the (EE) stage of P. berghei in murine hepatocytes and in two cell lines derived from the livers of BALB/c mice; one line produced from a primary hepatocyte culture and the other produced by fusion of mouse hepatocytes with a continuous rat liver line. Mature parasites were detected by microscopy and by DNA probe in both cell lines, each of which supported complete development of P. berghei liver stages and production of infectious merozoites. Since class I MHC antigens are present on the surface of primary hepatocytes and the mouse X rat hybrid line, these cells can be used to detect cytotoxic T cells against liver stage parasites.
伯氏疟原虫的红细胞外期（EE）已在人连续细胞系以及人源和大鼠源肝细胞的原代培养物中进行体外培养。尽管辐照子孢子诱导保护性免疫的主要实验模型是小鼠，但伯氏疟原虫尚未在原代小鼠肝细胞或小鼠连续细胞系中培养。因此，无法获得靶细胞来确定这些免疫小鼠是否产生细胞毒性T淋巴细胞（CTL），这些CTL能识别在I类主要组织相容性（MHC）抗原背景下肝细胞上表达的伯氏疟原虫抗原。我们报告了在小鼠肝细胞以及从BALB/c小鼠肝脏衍生的两种细胞系中培养伯氏疟原虫红细胞外期的方法；一种细胞系由原代肝细胞培养产生，另一种由小鼠肝细胞与大鼠肝脏连续细胞系融合产生。通过显微镜检查和DNA探针在两种细胞系中均检测到了成熟寄生虫，每种细胞系都支持伯氏疟原虫肝脏期的完全发育和感染性裂殖子的产生。由于I类MHC抗原存在于原代肝细胞和小鼠X大鼠杂交细胞系的表面，这些细胞可用于检测针对肝脏期寄生虫的细胞毒性T细胞。
