Miller G, Heston L
Yale J Biol Med. 1974;47(2):123-35.
Three continuous lymphoblastoid cell lines, 2 productive of nucleocapsids and 1 nonproductive line, were studied for their content of Epstein-Barr viral (EBV) antigens during transition from stationary to logarithmic phase growth. As a preliminary step, viable cells were separated from degenerating ones in discontinuous gradients of serum albumin. Viral capsid antigens were found in both living and dead cells of the 2 producer lines; however, complement fixing (CF) antigens and nuclear antigen were detected only in viable cell subpopulations. The content of antigen detectable in extracts of viable cells by complement fixation remained constant in replicating and resting cultures; further, all viable cells of the 3 lines demonstrated intranuclear antigen by anticomplement immunofluorescence in all stages of growth. In contrast, the proportion of cells with viral capsid antigen in the producer lines increased 7- to 24-fold following entry of resting populations into the phase of exponential growth.The results suggest that expression of viral capsid antigens is discontinuous and is initiated in response to events in log phase, possibly DNA synthesis or mitosis. Expression of the complement fixing and nuclear antigens in continuous in viable cells. These findings emphasize the intimate relationship of the CF and nuclear antigen to the transformed state and suggest that study of this antigen complex will shed light on the mechanisms of lymphocyte transformation by EBV.
研究了三种连续的淋巴母细胞系,其中两种能产生核衣壳,一种不能产生,观察它们在从静止期到对数生长期转变过程中 Epstein-Barr 病毒(EBV)抗原的含量。作为初步步骤,在血清白蛋白的不连续梯度中,将活细胞与退化细胞分离。在两种能产生病毒的细胞系的活细胞和死细胞中均发现了病毒衣壳抗原;然而,补体结合(CF)抗原和核抗原仅在活细胞亚群中检测到。通过补体结合在活细胞提取物中可检测到的抗原含量在复制培养和静止培养中保持恒定;此外,在生长的各个阶段,这三种细胞系的所有活细胞通过抗补体免疫荧光均显示出核内抗原。相比之下,静止群体进入指数生长期后,能产生病毒的细胞系中具有病毒衣壳抗原的细胞比例增加了 7 至 24 倍。结果表明,病毒衣壳抗原的表达是不连续的,并且是在对数期响应某些事件而启动的,可能是 DNA 合成或有丝分裂。补体结合抗原和核抗原在活细胞中的表达是持续的。这些发现强调了 CF 抗原和核抗原与转化状态的密切关系,并表明对这种抗原复合物的研究将有助于揭示 EBV 介导淋巴细胞转化的机制。