Augustine Nancy H, Pasi Brian M, Hill Harry R
Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84132, USA.
J Clin Lab Anal. 2007;21(5):265-70. doi: 10.1002/jcla.20182.
Lymphocyte proliferation in response to mitogens, phytohemagglutinin (PHA), concanavalin A, pokeweed, and/or specific antigens has been the method of choice for in vitro diagnosis of cell-mediated immune dysfunction. Recently, an assay to measure intracellular adenosine triphosphate (ATP) production in response to PHA has been developed that requires a shorter, overnight incubation. We compared a standard 5- to 7-day lymphocyte mitogen stimulation assay utilizing tritiated thymidine (3H-thy) incorporation to one in which ATP production in response to PHA by CD4-positive cells is measured in a luminometer that requires only 18-24 hr. A total of 20 patient samples suspected of having decreased cell-mediated immunity submitted for mitogen induced lymphocyte proliferation and 21 normal controls were tested in both assays. A comparison of these two methods has demonstrated that the screening ATP assay has a sensitivity at 24 hr of 100% in detecting decreased PHA induced lymphocyte proliferation at 5 days and a specificity of 85% in the samples obtained from normal controls. The data indicate that the ATP assay may be a useful screening tool for more rapid detection of blood samples with decreased cell-mediated immune responses. However, a positive screen should always be confirmed by 3H-thy uptake using mitogens and recall antigens like candida and tetanus.
淋巴细胞对丝裂原、植物血凝素（PHA）、刀豆球蛋白A、商陆以及/或者特定抗原的增殖反应一直是体外诊断细胞介导免疫功能障碍的首选方法。最近，已开发出一种用于测量PHA刺激下细胞内三磷酸腺苷（ATP）生成的检测方法，该方法只需较短的过夜孵育时间。我们将利用氚标记胸腺嘧啶核苷（³H-胸苷）掺入的标准5至7天淋巴细胞丝裂原刺激试验与一种在发光计中测量CD4阳性细胞对PHA反应时ATP生成的试验进行了比较，后者仅需18至24小时。对总共20份怀疑细胞介导免疫降低而提交进行丝裂原诱导淋巴细胞增殖检测的患者样本以及21份正常对照样本进行了这两种试验检测。这两种方法的比较表明，ATP筛选检测在24小时时检测5天时PHA诱导淋巴细胞增殖降低的敏感性为100%，在从正常对照获得的样本中的特异性为85%。数据表明，ATP检测可能是一种用于更快速检测细胞介导免疫反应降低的血样的有用筛选工具。然而，阳性筛选结果始终应用丝裂原和念珠菌、破伤风等回忆抗原来通过³H-胸苷摄取进行确认。
