Husain Zaheed, Alper Chester A, Yunis Edmond J, Dubey Devendra P
Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA.
Immunology. 2002 Jul;106(3):373-80. doi: 10.1046/j.1365-2567.2002.01444.x.
Human natural killer (NK) cells express several inhibitory and non-inhibitory NK receptors per cell. Understanding the expression patterns of these receptor genes in individual cells is important to understanding their function. Using a single-cell reverse transcription-polymerase chain reaction (RT-PCR) method, we analysed the expression of nine NK receptor genes in 38 resting CD56+ NK cells from peripheral blood of normal donors. We observed highly diverse patterns of receptor expression in these cells. No NK receptor is expressed universally in every CD56+ NK cell. The expressed receptor types per cell varied from two to eight. We specifically analysed the distribution of inhibitory (DL) and non-inhibitory (DS) killer immunoglobulin-like receptors (KIR). The frequency of individual receptor expression varied from 26% for 2DS2 to 68% for both 2DL1 and 2DL4. A comparison of the coexpression of DL and DS receptors showed a significant association in the expression of 2DL2 and 2DS2 (chi2=16.6; P<0.001) genes but no association between 2DL1 and 2DS1 or between 3DL1 and 3DS1 genes. Coexpression analysis of the 2DL1 and 2DL2 genes in 2DL4+ and 2DL4- cells showed a strong association in 2DL4+ but not in 2DL4- cells, suggesting a differential effect of the 2DL4 gene on the expression of 2DL1 and 2DL2 genes. Single-cell RT-PCR is a powerful tool to study multiple receptor gene expression ex vivo in individual NK cells and provides information about the expression pattern of KIR receptors that may suggest mechanisms of gene expression responsible for generation of the KIR repertoire.
人类自然杀伤（NK）细胞每个细胞表达多种抑制性和非抑制性NK受体。了解这些受体基因在单个细胞中的表达模式对于理解其功能很重要。我们使用单细胞逆转录聚合酶链反应（RT-PCR）方法，分析了来自正常供体外周血的38个静息CD56+NK细胞中9种NK受体基因的表达。我们在这些细胞中观察到高度多样的受体表达模式。没有一种NK受体在每个CD56+NK细胞中普遍表达。每个细胞表达的受体类型从2种到8种不等。我们专门分析了抑制性（DL）和非抑制性（DS）杀伤细胞免疫球蛋白样受体（KIR）的分布。单个受体表达的频率从2DS2的26%到2DL1和2DL4的68%不等。DL和DS受体共表达的比较显示2DL2和2DS2基因的表达有显著相关性（χ2=16.6；P<0.001），但2DL1和2DS1基因之间或3DL1和3DS1基因之间没有相关性。2DL4+和2DL4-细胞中2DL1和2DL2基因的共表达分析显示在2DL4+细胞中有很强的相关性，而在2DL4-细胞中没有，这表明2DL4基因对2DL1和2DL2基因的表达有不同的影响。单细胞RT-PCR是一种强大的工具，可用于在体外研究单个NK细胞中多种受体基因的表达，并提供有关KIR受体表达模式的信息，这可能提示负责KIR库产生的基因表达机制。
