T-lymphocyte colonies can be obtained from normal human peripheral blood and bone marrow, using a double layer technique which requires the simultaneous presence of peripheral blood irradiated leucocytes in the underlayer and phytohaemagglutinin in the overlayer (Foa CATOVSKY, Clin. Exp. Immunol., 36, 488, 1979). The absence of either of these two factors unables colony formation. In an attempt to improve the plating efficiency 15% autologous plasma was added to the leucocyte rich underlayer, or tested alone. The results demonstrated that both leucocytes and plasma alone show a similar colony promoting activity, giving rise respectively to 160 +/- 80.4 SD and 180 +/- 113.5 SD T-colonies. The combination of both factors produced a significant increase in growth, with an overall mean of 410 +/- 190.5 colonies. Increasing concentration of plasma (1%, 5%, 10%, 15%) gave rise to a dose-dependent increase in colony growth. Furthermore, the simultaneous presence of leucocytes and plasma in the underlayer enabled good colony formation with as little as 0.1 x 105 tes cells in the overlayer. These findings indicate that both human peripheral blood leucocytes and plasma possess T-colony stimulating activity, and that optimal growth is achieved with the combination of these two factors in the underlayer.
[Effect of human plasma on the in vitro formation of T-lymphocyte colonies]
FIERRO, Maria Teresa;
1982-01-01
Abstract
T-lymphocyte colonies can be obtained from normal human peripheral blood and bone marrow, using a double layer technique which requires the simultaneous presence of peripheral blood irradiated leucocytes in the underlayer and phytohaemagglutinin in the overlayer (Foa CATOVSKY, Clin. Exp. Immunol., 36, 488, 1979). The absence of either of these two factors unables colony formation. In an attempt to improve the plating efficiency 15% autologous plasma was added to the leucocyte rich underlayer, or tested alone. The results demonstrated that both leucocytes and plasma alone show a similar colony promoting activity, giving rise respectively to 160 +/- 80.4 SD and 180 +/- 113.5 SD T-colonies. The combination of both factors produced a significant increase in growth, with an overall mean of 410 +/- 190.5 colonies. Increasing concentration of plasma (1%, 5%, 10%, 15%) gave rise to a dose-dependent increase in colony growth. Furthermore, the simultaneous presence of leucocytes and plasma in the underlayer enabled good colony formation with as little as 0.1 x 105 tes cells in the overlayer. These findings indicate that both human peripheral blood leucocytes and plasma possess T-colony stimulating activity, and that optimal growth is achieved with the combination of these two factors in the underlayer.
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