Nooter K, Coolen J, Dubbes R, Zurcher C, Koch G, Bentvelzen P
J Gen Virol. 1979 Dec;45(3):711-21. doi: 10.1099/0022-1317-45-3-711.
Bone marrow of leukaemic patients, non-leukaemic patients and normal individuals were co-cultivated with the canine cell line A7573. These co-cultures were screened for retrovirus antigens by means of the indirect cytoplasmic immunofluorescence assay (IFA). Rabbit antisera directed against the major structural protein (p30) of woolly monkey (simian) sarcoma leukaemia virus (grown in human lymphoid cells) and Rauscher murine leukaemia virus were used for testing. After 2 months in culture, 6 of 17 co-cultures containing cells from leukaemic patients showed positive staining in the IFA with the anti-simian virus serum. In control dog cells fluorescence was never observed. Five of the six positive cultures were derived from leukaemic children. One of 12 co-cultures of the non-leukaemic group and one of nine normal bone marrow co-cultures were positive with the simian virus antiserum. None of the 38 co-cultures stained positive in the IFA with Rauscher virus antiserum. Absorption of the simian virus antiserum with calf serum or mouse mammary tumour virus had no dramatic effect in the IFA on positive control cells or on cells of a positive co-culture. However, absorption with purified simian virus (grown in rat cells) completely abolished these fluorescence reactions. The results provide evidence that simian sarcoma-leukaemia virus related information was present in the original bone marrow samples and that co-cultivation with permissive mammalian cells enabled the detection of virus footprints.
将白血病患者、非白血病患者和正常个体的骨髓与犬类细胞系A7573共同培养。通过间接细胞质免疫荧光测定法(IFA)对这些共培养物进行逆转录病毒抗原筛查。使用针对绒毛猴(猿猴)肉瘤白血病病毒(在人淋巴细胞中生长)的主要结构蛋白(p30)的兔抗血清和劳舍尔鼠白血病病毒进行检测。培养2个月后,17个含有白血病患者细胞的共培养物中有6个在IFA中用抗猿猴病毒血清显示阳性染色。在对照犬细胞中从未观察到荧光。6个阳性培养物中有5个来自白血病儿童。非白血病组的12个共培养物中有1个以及9个正常骨髓共培养物中有1个用猿猴病毒抗血清呈阳性。38个共培养物中没有一个在IFA中用劳舍尔病毒抗血清染色呈阳性。用小牛血清或小鼠乳腺肿瘤病毒吸收猿猴病毒抗血清对IFA中的阳性对照细胞或阳性共培养物的细胞没有显著影响。然而,用纯化的猿猴病毒(在大鼠细胞中生长)吸收完全消除了这些荧光反应。结果提供了证据,表明猿猴肉瘤 - 白血病病毒相关信息存在于原始骨髓样本中,并且与允许的哺乳动物细胞共同培养能够检测到病毒痕迹。