Park B H, Good R A
Proc Natl Acad Sci U S A. 1972 Feb;69(2):371-3. doi: 10.1073/pnas.69.2.371.
We have developed a new micromethod for testing phytohemagglutinin stimulation of human lymphocytes. 50 mul of peripheral blood is incubated at 37 degrees C directly, without cell separation, with 50 mul of tissue culture medium containing 50mug of phytohemagglutinin for 24 hr. Then, 0.5 muCi of tritiated thymidine in 50 mul of tissue culture medium is added and incubated for an additional 16 hr. After incubation, erythrocytes are selectively lysed in distilled water, and white blood cells are trapped quickly on a Millipore filter. The radioactivity of the filter is then assessed by a scintillation counter. The requirement of extremely small amounts of blood and a short incubation time are advantages. Further, this method provides a most useful evaluation of thymusdependent cell populations in man and experimental animals and, for the first time, this method provides the possibility of quantitating the deficiency of thymus-dependent cells.
我们已经开发出一种用于检测植物血凝素对人淋巴细胞刺激作用的新微量方法。50微升外周血在不进行细胞分离的情况下，于37℃直接与含有50微克植物血凝素的50微升组织培养基孵育24小时。然后，加入50微升含有0.5微居里氚标记胸腺嘧啶核苷的组织培养基，并再孵育16小时。孵育后，红细胞在蒸馏水中被选择性裂解，白细胞迅速截留在微孔滤膜上。然后用闪烁计数器评估滤膜的放射性。所需血量极少且孵育时间短是其优点。此外，该方法对人和实验动物的胸腺依赖性细胞群体提供了非常有用的评估，并且首次提供了定量胸腺依赖性细胞缺陷的可能性。
