Hillyer G V
Laboratory of Parasite Immunology and Pathology, University of Puerto Rico School of Medicine, San Juan.
Am J Trop Med Hyg. 1989 Dec;41(6):674-9. doi: 10.4269/ajtmh.1989.41.674.
Cells from adult Fasciola hepatica were fused with cells from a murine BALB/c myeloma Sp2 line. The hybrid cells were grown in HAT (hypoxanthine, aminopterin, and thymidine) medium, cloned and subcloned, and shown to express parasite antigen for 1 year after fusion. Expression of parasite antigen was demonstrated by the following: 2 histogram flow cytometric analyses, in which a population of hybrid cells in the population of 7 month cultured hybrid cells showed 57% more fluorescence when treated with an anti-F. hepatica serum followed by anti-rabbit immunoglobulin G coupled to fluorescein isothiocyanate as compared with the same hybrid cells washed and treated with normal rabbit serum; Sp2 myeloma cells treated with an anti-F. hepatica serum or normal rabbit serum followed by fluorescein-labeled anti-rabbit IgG had the same negative fluorescence; BALB/c mice immunized with PBS-washed cells from a subclone of these hybridomas developed anti-F. hepatica antibodies (shown by the Falcon assay screening test enzyme-linked immunosorbent assay); and antibodies recognized an F. hepatica antigenic polypeptide of 57,000 Mr in a Western immunoblot. These helminth:myeloma hybrids expressed murine host markers, further confirming the hybrid nature of this cell line. F. hepatica cells alone, like their Sp2 fusion partners, die in HAT supplemented medium by 9 days of culture. F. hepatica:Sp2 hybridomas have been grown continuously in HAT medium for greater than 1 year.
将成年肝片吸虫的细胞与小鼠BALB/c骨髓瘤Sp2系的细胞进行融合。杂交细胞在HAT（次黄嘌呤、氨基蝶呤和胸腺嘧啶核苷）培养基中培养，进行克隆和亚克隆，并显示在融合后1年内表达寄生虫抗原。通过以下方法证实了寄生虫抗原的表达：2次直方图流式细胞术分析，其中在培养7个月的杂交细胞群体中，与用正常兔血清洗涤并处理的相同杂交细胞相比，用抗肝片吸虫血清处理，然后用与异硫氰酸荧光素偶联的抗兔免疫球蛋白G处理的杂交细胞群体显示出多57%的荧光；用抗肝片吸虫血清或正常兔血清处理，然后用荧光素标记的抗兔IgG处理的Sp2骨髓瘤细胞具有相同的阴性荧光；用这些杂交瘤亚克隆的经PBS洗涤的细胞免疫的BALB/c小鼠产生了抗肝片吸虫抗体（通过福尔康检测筛选试验酶联免疫吸附测定显示）；并且抗体在蛋白质免疫印迹中识别出一种57000相对分子质量的肝片吸虫抗原多肽。这些蠕虫：骨髓瘤杂交细胞表达小鼠宿主标志物，进一步证实了该细胞系的杂交性质。单独的肝片吸虫细胞，就像它们的Sp2融合伙伴一样，在添加HAT的培养基中培养9天后死亡。肝片吸虫：Sp2杂交瘤已在HAT培养基中连续培养超过1年。
