McCoy J L, Dean J H, Herberman R B
J Immunol Methods. 1977;15(4):355-71. doi: 10.1016/0022-1759(77)90097-7.
Direct and indirect agarose microdroplet migration inhibition assays were performed with log dilutions (50--5 X 10(-8) microgram/ml) of soluble purified protein derivative (PPD) of tuberculin and leukocytes (4 X 10(5)/droplet) from PPD skin test positive or negative individuals. Some tests were run in parallel with the capillary tube method. Inhibition of migration of leukocytes from 9/11 PPD skin test positive donors studied was observed in direct tests when employing PPD doses ranging from 1--50 microgram/ml PPD. Inhibition of migration of leukocytes from some PPD skin reactive donors was obtained with as little as 5 X 10(-3)--5 X 10(-7) microgram/ml (i.e., 5 nanograms to 0.5 picograms). Some inhibition of leukocyte migration was seen with skin test negative donors (presumably toxicity) with the higher doses of PPD (50 microgram/ml), but generally little inhibition was observed with these donors at lower doses. Comparative experimetns of the agarose micromethod and the capillary tube technique indicated that the agarose assay was more sensitive in that it could detect reactivity with 2--4 logs lower antigen concentration. Indirect assays using supernatants of cultures of PPD triggered mononuclear cells and indicator granulocytes in agarose droplets demonstrating that a lymphokine (presumable leukocyte inhibitory factor) was being produced and suggested that cell-mediate immune (CMI) reactions were operative. The results indicate the usefulness of this technique in the sensitive-detection of CMI against such antigens as soluble PPD. The assay should prove useful in quantitative assessment of cell-mediated reactivity by using a wide range of antigen concentrations and the leukocytes from as little as 2--5 ml of blood.
采用结核菌素可溶性纯化蛋白衍生物（PPD）的对数稀释液（50 - 5×10⁻⁸微克/毫升）以及PPD皮肤试验阳性或阴性个体的白细胞（4×10⁵/微滴），进行了直接和间接琼脂糖微滴迁移抑制试验。一些试验与毛细管法平行进行。在直接试验中，当使用1 - 50微克/毫升PPD剂量时，观察到9/11名接受研究的PPD皮肤试验阳性供体的白细胞迁移受到抑制。对于一些PPD皮肤反应性供体，低至5×10⁻³ - 5×10⁻⁷微克/毫升（即5纳克至0.5皮克）的PPD剂量就能抑制白细胞迁移。皮肤试验阴性供体在较高剂量PPD（50微克/毫升）时，白细胞迁移也有一定抑制（可能是毒性作用），但在较低剂量时，这些供体通常很少出现抑制现象。琼脂糖微量法和毛细管技术的对比实验表明，琼脂糖试验更敏感，因为它能检测到抗原浓度低2 - 4个对数时的反应性。使用PPD刺激的单核细胞培养上清液和琼脂糖微滴中的指示粒细胞进行的间接试验表明，正在产生一种淋巴因子（可能是白细胞抑制因子），提示细胞介导免疫（CMI）反应起作用。结果表明该技术在灵敏检测针对可溶性PPD等抗原的CMI方面有用。该试验通过使用广泛的抗原浓度以及仅2 - 5毫升血液中的白细胞，在定量评估细胞介导反应性方面应会很有用。
