Purification of human T and B cells by a discontinuous density gradient of percoll.
A simple technique for isolating human peripheral B and T cells is described. This method is based on the different mobility of cells in a discontinuous density gradient of Percoll. A population with a mean of 72.8% of surface stable Ig (B cells) is found in the top layer (fraction I) whereas T cells with less than 3% of Ig are distributed in the two lower layers (fractions II and III). B cells obtained by this method were highly viable and reacted strongly with anti-DRw specific sera, whereas T cells did not respond. The clear-cut difference between positive and negative reactions makes this technique very suitable for DRw typing. The proliferative response to PHA is decreased in fractions I and III while greatly increased in fraction II. The different mitogenic responses in the two fractions containing T cells probably represent two functionally distinct T cell subsets. Cells from all fractions are less activated than unseparated cells when Con A and PWM are used as mitogen. The technique is very rapid, avoids the interaction of cells with their specific receptors and gives both T and B cells in one step, all advantages over other methods commonly used.
D000961:Antilymphocyte Serum / Q000494:pharmacology; D001402:B-Lymphocytes; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003102:Colloids; D006801:Humans; D008213:Lymphocyte Activation; D011205:Povidone; D011947:Receptors, Antigen, B-Cell; D012397:Rosette Formation; D012822:Silicon Dioxide; D013601:T-Lymphocytes
D000961:Antilymphocyte Serum; D003102:Colloids; D011947:Receptors, Antigen, B-Cell; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone