Quantitation Of Viral Reproduction

没有可靠的通用方法来量化一个普通病毒复制的时间。从宏观上讲，一种“普通”病毒在一种文化中的产仔数和繁殖时间是多少？该项目开发了数学和实验技术，用于确定单个母体病毒粒子附着在细胞上时的子体病毒粒子的平均数、子体的平均周期时间以及病毒周期时间的标准差(传播)。这些测定可以通过流式细胞仪实验进行。靶细胞用脂类染料染色，病毒在它们的贴壁阶段通过与被染色的0代细胞孵育而同步。通过收集感染标记物(在实际实验中，这将是HIV中的gp41)的数据，无论是在第0代染色的细胞中还是在随后添加的未染色的细胞中，并与受感染的“第0代”细胞孵育，平均病毒的复制都可以作为积分方程式的解来确定。原则上，在将一群同类动物放生到野外并进行几次种群计数后，同样的方法可以用于计算一只动物的窝大小和繁殖周期。然而，在医学方面，这项工作的目的是改进各种艾滋病毒疗法的量化及其对艾滋病毒繁殖的抑制作用。计算机程序现在已经就位，一篇理论论文已经发表，实验方案现在已经实施，数据目前正在分析中。J.L.Spouge和S.P.Layne：“一种同时确定病毒有效爆发大小和周期时间的实用方法”(1999)。娜塔莉。阿卡德。SCI。美国96：7017-7022

There is no dependable general way of quantifying the times at which an average virus reproduces. In macroscopic terms, what are the litter sizes and breeding times of an "average" virus in a culture? This project develops mathematical and experimental techniques for determining the average number of daughter virions from a single mother virion, when these daughters attach to their cell, the average cycle time for the daughters, and the standard deviation (spread) of the viral cycle time. These determinations can be made from flow cytometry experiments. Target cells are stained with lipid dyes, and viruses syncronized at their attachment phase by incubating them with the stained "generation 0" cells. By collecting data on a marker of infection (in actual experiments, this will be gp41 in HIV), both in generation 0 stained cells and unstained cells that are added later and incubated with the infected "generation 0" cells, the reproduction of the average virus can be determined as the solution of an integral equation. The same methods can in principle be employed in calculating litter sizes and breeding cycle times of an animal, after a homogeneous group of the animals is released into the wild and several populaton counts taken. In the medical context, however, this work is intended to improve the quantitation of various HIV therapies and their inhibitory effects on HIV reproduction. Computer programs are now in place, a theoretical paper has been published, experimental protocols have now been carried out, and the data are currently being analyzed. J. L. Spouge and S. P. Layne "A practical method for simultaneously determining the effective burst sizes and cycle times of viruses" (1999) Proc. Natl. Acad. Sci. USA 96 : 7017-7022