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首页> 外文期刊>Journal of Virology >Autoradiographic method for detection of lactate dehydrogenase-elevating virus-infected cells in primary mouse macrophage cultures.
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Autoradiographic method for detection of lactate dehydrogenase-elevating virus-infected cells in primary mouse macrophage cultures.

机译：原发性小鼠巨噬细胞培养物中乳酸脱氢酶升高病毒感染细胞的放射造影方法。

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Peritoneal cells from starch-injected Swiss mice were propagated in plastic petri dishes and on cover slips in a mouse L-cell-conditioned medium for 12 to 24 h and then infected with various multiplicities of lactate dehydrogenase-elevating virus (LDV). Over 95% of the cells in these cultures phagocytosed latex particles and were, therefore, considered macrophages. Infected and mock infected macrophage cultures were supplemented with [3H]uridine at various times after infection and with actinomycin D 30 min before addition of the [3H]uridine. After 1 or 2 h of further incubation, plate cultures were analyzed for LDV-specific RNA, and cover slip cultures were investigated by autoradiography. Other cultures were labeled in the absence of actinomycin D, and the culture fluid was analyzed for labeled LDV. There was a good correlation between the production of LDV-specific RNA and LDV and the number of heavily labeled cells in these cultures. The labeled cells in these cultures. The labeled cells, therefore, were equated with productively infected cells. Only a maximum of about 20% of the macrophages, however, became heavily labeled regardless of the multiplicity of infection or the time, after infection, at which the cells were exposed to [3H]uridine. Only background labeling was observed in the remainder of the cells and in mock-infected cells treated with actinomycin D. The highest proportion of labeled cells was observed when the cells were infected with a multiplicity of infection of about 2,000 mouse infectious units per cell and labeled from 6 to 8 h after infection. Thereafter, the proportion of productively infected cells decreased progressively, concomitant with a decrease in the amounts of viral specific RNA and of LDV produced by the cultures. The results indicate that the majority of the macrophages in primary macrophage cultures do not support LDV replication. Their nonpermissiveness may depend on the physiological state of the cells or reflect the presence of subpopulations of macrophages, but no morphological differences between productively infected an uninfected cells were detectable.
机译：来自淀粉注入的瑞士小鼠的腹膜细胞在塑料培养皿中和小鼠L细胞条件培养基中的盖子上繁殖12至24小时，然后用各种多重乳酸脱氢酶升降病毒（LDV）感染。这些培养物中超过95％的细胞吞噬胶乳颗粒，因此被认为是巨噬细胞。感染和嘲弄感染的巨噬细胞培养物在感染后的不同时间补充有[3H]尿苷，并在加入[3H]尿苷之前30分钟，凝集霉素D 30分钟。在进一步孵育1或2小时之后，分析平板培养物用于LDV特异性RNA，通过放射自显影研究了覆盖滑脂。在没有放线菌素D的情况下标记其他培养物，并分析培养物用于标记的LDV。 LDV特异性RNA和LDV的生产与这些培养物中的重标记细胞数量之间存在良好的相关性。这些培养物中的标记细胞。因此，标记的细胞与有效感染的细胞等同。然而，只有大约20％的巨噬细胞，无论多种感染或时间，感染后，细胞暴露于[3H]尿苷中的时间，大约20％的巨噬细胞变得严重标记。在细胞的其余部分中观察到背景标记，并在用放线菌素D处理的模拟感染细胞中。当细胞感染细胞每种细胞约2,000只小鼠感染单位的多种感染和标记时，观察到最高的标记细胞的比例。感染后6至8​​小时。此后，高效细胞的比例逐渐减少，伴随着病毒特异性RNA和由培养物产生的LDV的量减少。结果表明，主要巨噬细胞培养物中的大多数巨噬细胞不支持LDV复制。它们的非肿注可能取决于细胞的生理状态或反映巨噬细胞群的存在，但没有能量感染的形态学差异是可检测的。

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