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胰蛋白酶对仙台病毒在组织培养细胞中生长的作用。I. 胰蛋白酶对L细胞携带的仙台病毒的直接作用恢复其对L细胞的感染性。

Trypsin action on the growth of Sendai virus in tissue culture cells. I. Restoration of the infectivity for L cells by direct action of tyrpsin on L cell-borne Sendai virus.

作者信息

Homma M

出版信息

J Virol. 1971 Nov;8(5):619-29. doi: 10.1128/JVI.8.5.619-629.1971.

Abstract

Sendai virus grown in fertile eggs (egg Sendai) infects L cells in which the synthesis of L Sendai (grown in L cells) occurs by the one-step mechanism. L Sendai is not infectious for L cells when tested by the tube titration method although it is infectious for chick embryos. When L cells infected with egg Sendai were dispersed by trypsin and plated on a monolayer culture of L cells, the viral agents spread to the adjacent recipient cells in which the synthesis of L Sendai occurred. The newly infected L cells became infectious for L cells again by trypsin treatment. Kinetic experiments suggested that the target of trypsin is the mature virus, of L Sendai nature, just budding from the L-cell surface. By using an immunofluorescent cell-counting technique, recovery of the infectivity of L Sendai for L cells due to a direct enzymatic action of trypsin was demonstrated. Under the optimal condition, the infectivity increased 1,000-fold for L cells and 10-fold for chick embryos, and both the titers could favorably be compared. No increasing effect of trypsin was observed on the infectivity of egg Sendai. Density centrifugation studies revealed a difference between egg Sendai and L Sendai in the density. Trypsin treatment which induced the maximal enhancement of L Sendai infectivity did not affect both the densities, showing that variations of Sendai virus in the infectivity for L cells and in the density are independent types of host-controlled modification.

摘要

在受精鸡蛋中培养的仙台病毒(鸡蛋仙台病毒)可感染L细胞,在L细胞中,L仙台病毒(在L细胞中培养)通过一步机制进行合成。尽管L仙台病毒对鸡胚具有感染性,但通过试管滴定法检测时,它对L细胞无感染性。当用胰蛋白酶将感染了鸡蛋仙台病毒的L细胞分散,并接种到L细胞的单层培养物上时,病毒因子会扩散到相邻的受体细胞中,在这些细胞中会发生L仙台病毒的合成。新感染的L细胞经胰蛋白酶处理后,再次对L细胞具有感染性。动力学实验表明,胰蛋白酶的作用靶点是刚从L细胞表面出芽的、具有L仙台病毒性质的成熟病毒。通过使用免疫荧光细胞计数技术,证实了由于胰蛋白酶的直接酶促作用,L仙台病毒对L细胞的感染性得以恢复。在最佳条件下,L仙台病毒对L细胞的感染性增加了1000倍,对鸡胚的感染性增加了10倍,且两种滴度具有良好的可比性。未观察到胰蛋白酶对鸡蛋仙台病毒的感染性有增强作用。密度离心研究揭示了鸡蛋仙台病毒和L仙台病毒在密度上的差异。诱导L仙台病毒感染性最大增强的胰蛋白酶处理并未影响两者的密度,这表明仙台病毒在对L细胞的感染性和密度方面的变化是宿主控制修饰的独立类型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c75/376239/2417954384e6/jvirol00287-0019-a.jpg

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