Westera Liset, Zhang Yan, Tesselaar Kiki, Borghans José A M, Macallan Derek C
Department of Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
Methods Mol Biol. 2013;979:107-31. doi: 10.1007/978-1-62703-290-2_10.
Humans have a remarkable ability to maintain relatively constant lymphocyte numbers across many decades, from puberty to old-age, despite a multitude of infectious and other challenges and a dramatic decline in thymic output. This phenomenon, lymphocyte homeostasis, is achieved by matching the production, death, and phenotype transition rates across a network of varied lymphocyte subpopulations. Understanding this process in humans depends on the ability to measure in vivo rates of lymphocyte production and loss. Such investigations have been greatly facilitated by the advent of stable isotope labeling approaches, which use the rate of incorporation of a tracer into cellular DNA as a marker of cell division. Two labeling approaches are commonly employed, one using deuterium-labeled glucose and the other using deuterium-labeled water, also known as heavy water ((2)H(2)O). Here we describe the application of these two labeling techniques for measurement of human in vivo lymphocyte kinetics through the four phases of investigation: labeling, -sampling, analysis, and interpretation.
从青春期到老年的数十年间，尽管面临众多感染及其他挑战，且胸腺输出急剧下降，但人类仍具有非凡的能力，能够使淋巴细胞数量保持相对恒定。这种现象，即淋巴细胞稳态，是通过在不同淋巴细胞亚群网络中匹配产生、死亡和表型转变速率来实现的。了解人类的这一过程依赖于测量体内淋巴细胞产生和损失速率的能力。稳定同位素标记方法的出现极大地推动了此类研究，该方法利用示踪剂掺入细胞DNA的速率作为细胞分裂的标志物。通常采用两种标记方法，一种使用氘标记的葡萄糖，另一种使用氘标记的水，也称为重水（(2)H(2)O）。在此，我们描述这两种标记技术在通过四个研究阶段测量人类体内淋巴细胞动力学方面的应用：标记、采样、分析和解释。
