McFadden R G
Lawson Research Institute, London, Ontario, Canada.
J Clin Lab Immunol. 1988 Aug;26(4):201-8.
The application of mononuclear cell populations to a nylon wool (NW) column is a common early procedure in the selection of T lymphocytes for further study. The technique as presently employed in various laboratories does not appear to be standardized, and surprisingly little is known about the effects of physicochemical alterations and biologic mediators on the interaction of lymphocytes with the NW substrate. In this study the factors controlling the adherence of rat splenocytes to NW were examined. NW adherence was shown to be independent of loaded cell concentration, column packing or pH, but was very dependent on NW column size, wash volume, incubation time and ambient temperature. The addition of protein to media did not alter lymphocyte NW adherence, and the interaction did not appear to depend on intact cellular glycolysis or protein synthesis or on microtubular or microfilament function. Because of the theoretical importance of surface adherence to cell motility, the effects of various agents that alter lymphocyte migration were tested in the lymphocyte NW assay. Of the positive chemokinetic factors tested, only casein altered (decreased) NW adherence. Of the negative chemokinetic principles tested only the human lymphokine LyMIF altered (increased) NW adherence. The studies show that the NW-nonadherent cell pool may be a heterogeneous population depending on the physical conditions of the assay, and the NW adherence of rat splenocytes is not an all-or-none phenomenon but can be altered by physical and biological factors. This makes the standardization of the assay of critical importance, particularly if one wishes to compare results of subsequent experiments between laboratories.
将单核细胞群体应用于尼龙毛（NW）柱是筛选T淋巴细胞以便进一步研究的常见早期步骤。目前各实验室采用的这项技术似乎并未标准化，而且令人惊讶的是，对于物理化学改变和生物介质对淋巴细胞与NW底物相互作用的影响知之甚少。在本研究中，对控制大鼠脾细胞与NW黏附的因素进行了检测。结果表明，NW黏附与加载细胞浓度、柱填充或pH无关，但非常依赖于NW柱的大小、洗涤体积、孵育时间和环境温度。向培养基中添加蛋白质并不会改变淋巴细胞与NW的黏附，而且这种相互作用似乎并不依赖于完整的细胞糖酵解或蛋白质合成，也不依赖于微管或微丝功能。由于表面黏附对细胞运动具有理论重要性，因此在淋巴细胞NW试验中测试了各种改变淋巴细胞迁移的试剂的作用。在所测试的正向化学动力学因子中，只有酪蛋白改变（降低）了NW黏附。在所测试的负向化学动力学因子中，只有人淋巴因子LyMIF改变（增加）了NW黏附。这些研究表明，根据试验的物理条件，NW非黏附细胞池可能是一个异质群体，并且大鼠脾细胞与NW的黏附不是全或无的现象，而是可以被物理和生物学因素改变。这使得该试验的标准化至关重要，特别是如果有人希望比较不同实验室后续实验的结果。
