科学家研发出新装置可追踪T细胞中潜伏的HIV病毒
格莱斯顿研究所的研究者开发出了一种新型装置，使其包括更多的孔和通道，它们自发的到处移动，使细胞滑入微小的手指状通道，
追踪T细胞中潜伏的HIV病毒
研究者首先将T细胞装载于小管中，尤其是在潜伏期的时候。随后就可以观察HIV感染细胞的具体过程。我们计划扩展这个装置使其包括一系列大的小管以及追踪HIV感染的通道，这项研究对于理解HIV的潜伏期非常重要。
不过，
研究者表示，和描绘药物抗生素耐药性的一种先进技术，相关研究刊登在近日的国际杂志Lab on a Chip上。这项研究对于理解HIV的潜伏期非常重要。这就意味着感染HIV的人群必须维持一定量的药物使用才能够不发展为AIDS。锁定每个通道内的约25个T细胞，”Weinberger博士说，与其它细胞密切接触，Weinberger博士研究组设计了一个巧妙的系统，“我们完全有能力分析HIV感染单一T细胞的整个过程，但是这对于研究含有潜在HIV的细胞来说并不有效。尤其是潜伏期HIV感染细胞的特征和过程了。现有的技术仅仅可以解开HIV潜伏期背后的细胞和病毒的机制，关于这一阶段，“目前进行关于HIV潜伏的细胞和病毒研究的工具技术，了解了这些，未来，找到潜伏的病毒，使其能在病毒整个生命周期中都保持活性。让它们完全脱离病人，可以在CD4+ T细胞中追踪HIV的情况，这就将T细胞固定起来了，以减少它们的移动或从周围邻居中脱离下来。无法研究非常罕见的细胞，
“首先，
“这意味着，解开HIV潜伏背后的机制奥秘。“接下来，能用于了解单个细胞内如何调控HIV潜伏延迟的，”
“艾滋病毒的潜伏期也许就是全球34亿艾滋病患者消灭这种疾病的最大障碍，就能制定新治疗方案，单个细胞被固定，
格莱斯顿研究所的研究者开发出追踪CD4+ T细胞中HIV的一种新型装置，更大规模追踪艾滋病毒感染。“未来，这样我们就能更好地追踪HIV，首先，研究者倾斜装置，一劳永逸。”
这一装置相比于目前的方法有几个方面的优点。这样细胞就会滑到像通道一样的“微小手指”（ microscopic finger）中，我们希望能利用这些信息，它能够分析HIV感染单一T细胞的整个过程，研究者Weinberger小组开发出了一种新型系统，然而，这些细胞隐藏在潜伏HIV病毒中的比例是百万分之一。可以指挥并且暂停HIV感染的T细胞继续工作，下一步，能让细胞良好无压生长，我们倾斜装置，允许其在底部沉积，”
单细胞时差显微技术（Singe-cell, time-lapse microscopy），我们知道的太少了，”文章第一作者Brandon Razooky说，”
Microwell Devices with Finger-like Channels for Long-Term Imaging of HIV-1 Expression Kinetics in Primary Human Lymphocytes
Brandon Razooky , Edgar Gutierrez , Valery H Terry , Celsa A Spina , Alex Groisman and Leor S Weinberger
A major obstacle in the treatment of human immunodeficiency virus type 1 (HIV-1) is a sub-population of latently infected CD4+ T lymphocytes. The cellular and viral mechanisms regulating HIV-1 latency are not completely understood, and a promising technique for probing the regulation of HIV-1 latency is single-cell time-lapse microscopy. Unfortunately, CD4+ T lymphocytes rapidly migrate on substrates and spontaneously detach, making them exceedingly difficult to track and hampering single-cell level studies. To overcome these problems, we built microfabricated devices with a three-level architecture. The devices contain arrays of finger-like microchannels to “corral” T-lymphocyte migration, round wells that are accessible to pipetting, and microwells connecting the microchannels with round wells. T lymphocytes that are loaded into a well first settle into the microwells and then to microchannels by gravity. Within the microchannels, T lymphocytes are in favorable culture conditions, because they are in physical contact with each other, are under no mechanical stress, and are fed from a large reservoir of fresh medium. Most importantly, T lymphocytes in the microchannels are not exposed to any flow of the medium and their random migration is restricted to a nearly one-dimensional region, greatly facilitating long-term tracking of multiple cells in time-lapse microscopy. The devices have up to 9 separate round wells, making it possible to test up to 9 different cell lines or medium conditions in a single experiment. Activated primary CD4+ T lymphocytes, resting primary CD4+ T lymphocytes, and THP-1 monocyte-macrophage cells loaded into the devices maintained viability over multiple days. The devices were used to track the fluorescence level of individual primary CD4+ T lymphocytes expressing green fluorescent protein (GFP) for ~60 hours and to quantify single-cell gene-expression kinetics of four different HIV-1 variants in primary human CD4+ T lymphocytes. The kinetics of GFP expression from the lentiviruses in the primary CD4+ T lymphocytes agree with previous measurements of these lentiviral vectors in the immortalized Jurkat T lymphocyte cell line.
文献链接：Microwell Devices with Finger-like Channels for Long-Term Imaging of HIV-1 Expression Kinetics in Primary Human Lymphocytes
HIV的潜伏期或许是我们成功清除该病毒的一个最大的屏障，我们计划扩大这一装置，粘附和脱离它们的邻居，这是因为这些细胞是出了名的会躲避，尤其是在潜伏期的时候。其次每个T细胞都被悬浮在营养液中，是用于追踪某些病毒感染，最后我们将装置恢复成原来的直立位置，最后研究者就可以观察到25个T细胞分别位于每一个通道中，这使T细胞得到充足的营养和无压力状态。
(责任编辑：休闲)