Geczy C L, Meyer P A
J Immunol. 1982 Jan;128(1):331-6.
Human mononuclear leukocytes generate cell-bound procoagulant activity (LPCA) after incubation with an antigen (mumps or tuberculin) to which the donor was previously sensitized. An inhibitor of coagulation appears to be liberated into the extracellular culture fluid during incubation of leukocytes with the sensitizing antigen. Removal of this activity before measuring LPCA resulted in a reliable test that correlated directly with delayed skin reactivity. The assay was particularly sensitive in that cells from weakly sensitized donors who reacted only to high doses of tuberculin (100 TU) in the delayed skin tests produced detectable LPCA in vitro. By contrast cells from weakly sensitized donors did not react to PPD in the lymphocyte blast transformation test or the direct macrophage migration inhibition factor test. The LPCA assay correlated closely with the blast transformation and MIF tests in which cells were used from more strongly sensitized donors who reacted in skin tests with lower doses of tuberculin (1 or 10 TU). The assays were antigen-specific in that cells from donors sensitive to mumps antigen but not to tuberculin reacted only to mumps antigen in vitro. The assay was extremely reproducible; cells from individual donors reacted to the same extent over a period of 8 mo). We propose that the assay system reported here represents an improved method for the measurement of cell-mediated immunity in vitro because it requires fewer donor cells, is technically simpler, and is more sensitive than previously described methods.
人单核白细胞在与供体先前致敏的抗原(腮腺炎或结核菌素)孵育后会产生细胞结合促凝活性(LPCA)。在白细胞与致敏抗原孵育期间,一种凝血抑制剂似乎会释放到细胞外培养液中。在测量LPCA之前去除这种活性可得到一种可靠的检测方法,该方法与迟发型皮肤反应直接相关。该检测方法特别灵敏,因为在迟发型皮肤试验中仅对高剂量结核菌素(100 TU)有反应的弱致敏供体的细胞在体外能产生可检测到的LPCA。相比之下,弱致敏供体的细胞在淋巴细胞增殖试验或直接巨噬细胞移动抑制因子试验中对结核菌素纯蛋白衍生物(PPD)无反应。LPCA检测与增殖试验和巨噬细胞移动抑制因子试验密切相关,在这些试验中使用的是来自更强致敏供体的细胞,这些供体在皮肤试验中对较低剂量的结核菌素(1或10 TU)有反应。这些检测方法具有抗原特异性,即对腮腺炎抗原敏感但对结核菌素不敏感的供体的细胞在体外仅对腮腺炎抗原产生反应。该检测方法具有极高的可重复性;来自个体供体的细胞在8个月的时间内反应程度相同。我们认为,本文报道的检测系统代表了一种改进的体外细胞介导免疫测量方法,因为它所需的供体细胞较少,技术上更简单,且比先前描述的方法更灵敏。