Abstract
Lymphocyte blasttransformation by phytohemagglutinin (PHA) is thought to show the cellular immunological capacity of individuals. There are several problems with the evaluation of peripheral lymphocyte blasttransformation by the method which has heretofore been used.
For example, it is uncertain as to how many percent of initial lymphocytes transform to blastic cells within 72 hours, and as to what differenciation exists between the lymphocytes which would transform to blastic cells and those which would not transform; it is difficult to distinguish the relatively small blastic lymphocytes from non-blastic cells; and there is time differenciation among various lymphocytes to commence blastic transformation. To clarify these points, PHA lymphocyte culture added with small dose of colchicine (0.4γ/ml) was prepared. About 35% of initial lymphocytes obtained from healthy persons transformed to blastic cells within 72 hours by our method.
Using chromosome method with H3-thimidine labeled lymphocytes, it was confirmed that the lymphocytes in the conventional culture undergo mitosis 1 or 2 times within 72 hours. The addition of colchicine to the culture is to prevent such mitosis of cultured lymphocytes.
Peripheral lymphocytes of chronic lymphocytic leukemia, of malignant lymphoma with or without increased abnormal lymphocytes, and of filaria showed lower percentage of blasttransformation and the tendency of delayed reaction to PHA.
It seems better to utilize the number of H3-thimidine labeled cells plus cells of metaphase by our method to compare the degree of PHA blasttransformation, because these cells can be clearly distinguished.
Normal and CLL human peripheral lymphocytes could survive for 28 days in cell culture in the presence of small dose (subthreshold) of PHA. But peripheral lympocytes of leukemic malignant lymphoma disappeared from the culture of the same condition.
