Quantitation Of Viral Reproduction

没有可靠的通用方法来量化病毒平均繁殖的时间。从宏观上看，培养物中“平均”病毒的窝仔数和繁殖时间是多少？该项目开发了数学和实验技术，用于确定来自单个母体病毒体的子病毒体的平均数量，当这些子病毒体附着在它们的细胞上时，子病毒体的平均周期时间，以及病毒周期时间的标准偏差（扩散）。这些测定可以由流式细胞术实验进行。用脂质染料对靶细胞进行染色，并通过将病毒与染色的“0代”细胞一起孵育使病毒在其附着期同步。通过收集第0代染色细胞和随后加入并与感染的“第0代”细胞一起孵育的未染色细胞中感染标志物（在实际实验中，这将是HIV中的gp41）的数据，可以将平均病毒的繁殖确定为积分方程的解。原则上，在将同类动物群释放到野外并进行几次种群计数后，可以采用相同的方法计算动物的产仔数和繁殖周期。然而，在医学方面，这项工作旨在改善各种艾滋病毒疗法的定量及其对艾滋病毒繁殖的抑制作用。计算机程序现已到位，理论论文已发表，初步实验数据已分析。我们目前正在用新的细胞染色方法改进实验方案，并制定了进行实验和分析的时间表。 目前这一轮的实验失败是由于与基本理论无关的技术原因，该项目被搁置，直到有足够的实验资源可用。

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, preliminary experimental data have been analyzed. We are currently improving the experimental protocols with new cell staining methods, and a timetable for doing the experiments and analysis is in place. The current round of experiments failed for technical reasons unrelated to the underlying theory, and the project is on hold until adequate experimental resources are available.