Correlation between morphological blastformation rate and functional 3H-thymidine uptake in mixed lymphocyte culture in the presence of PHA.

By dividing at random 14 normal persons into 7 pairs of two individuals each, lymphocytes were isolated from their peripheral blood and taking one of the pairs as stimulating cells or antigens and the others as responding cells, mixed lymphocyte culture (MLC) was carried out. As for the method of MLC we used our MLC method of unidirectional mixed culture with a small amount of lymphocytes in additition of 1% (v /v) PHA.M and cultured for three days, and a widely used conventional method in which 3H.thymidine uptake was the parameter of the blastformation rate and cultured for seven days. In comparing the results of these two groups of MLC the data in six experiments out of the seven coincided. Namely, with 5x 104 cells each of stimulating cell group and responding cell group, it is possible to achieve satisfactory MLC, the culture can be done only for three days without requiring any special technique and the results can be readily evaluated. Therefore, MLC by our simple method would yield satisfactory results in clinics. ∗PMID: 4280235 [PubMed indexed for MEDLINE] Copyright c ©OKAYAMA UNIVERSITY MEDICAL SCHOOL Acta Merl. Okayama 28, 253-257 (1974) CORRELATION BETWEEN MORPHOLOGICAL BLASTFORMATION RATE AND FUNCTIONAL 3H.THYMIDINE UPTAKE IN MIXED LYMPHOCYTE CULTURE IN THE PRESENCE OF PHA Kunzo ORITA, Hiroaki MIWA and Shoken KANEDA Department oj Surgery, Okayama University Medical School, Okayama, Japan (Director: Proj. S. Tanaka) Received jor publication, January 17, 1974 Abstract: By dividing at random 14 normal persons into 7 pairs of two individuals each, lymphocytes were isolated from their peripheral blood and taking one of the pairs as stimulating cells or antigens and the others as responding cells, mixed lymphocyte culture (MLC) was carried out. As for the method of MLC we used our MLC method of unidirectional mixed culture with a small amount of lymphocytes in additition of 1% (v /v) PHA.M and cultured for three days, and a widely used conventional method in which 3H.thymidine uptake was the parameter of the blastformation rate and cultured for seven days. In comparing the results of these two groups of MLC the data in six experiments out of the seven coincided. Namely, with 5x 104 cells each of stimulating cell group and responding cell group, it is possible to achieve satisfactory MLC, the culture can be done only for three days without requiring any special technique and the results can be readily evaluated. Therefore, MLC by our simple method would yield satisfactory results in clinics. By dividing at random 14 normal persons into 7 pairs of two individuals each, lymphocytes were isolated from their peripheral blood and taking one of the pairs as stimulating cells or antigens and the others as responding cells, mixed lymphocyte culture (MLC) was carried out. As for the method of MLC we used our MLC method of unidirectional mixed culture with a small amount of lymphocytes in additition of 1% (v /v) PHA.M and cultured for three days, and a widely used conventional method in which 3H.thymidine uptake was the parameter of the blastformation rate and cultured for seven days. In comparing the results of these two groups of MLC the data in six experiments out of the seven coincided. Namely, with 5x 104 cells each of stimulating cell group and responding cell group, it is possible to achieve satisfactory MLC, the culture can be done only for three days without requiring any special technique and the results can be readily evaluated. Therefore, MLC by our simple method would yield satisfactory results in clinics. In performing organ transplantation recently the mixed lymphocyte culture (MLC) has come to occupy an important place along with HL-A typing. When lymphocytes from two different individuals are cultured together, the percentage of cells transformed into a blast type is inversely proportinal to the tissue compatibility of the two individuals. This MLC has a very highly significant value in predicting the clinical results of a renal allograft. We have already demonstrated that the unidirectional lymphocyte mixed culture of a small amount in the presence of PHA can predict fairly well the potentials of H·2 or non-H-2 histoincompatibility as well as the survival time of skin or kidney allograft (1-4). In the present investigation we conducted MLC by the 3H-thymidine incorporation, the method now widely used the world over (5), conventional method, and obtained the data coinciding with those obtained by our MLC with PHA and report here our findings.