Fractionation of human T lymphocytes on the basis of their high, medium and low SRBC-rosette-forming affinity: efficiency of the method

Abstract

Under optimal saturating conditions at SRBC/lymphocyte ratio 100:1 26 +/- 1.2% of human peripheral blood lymphocytes isolated from defibrinated blood form rosettes without incubation, 48 +/- 1.8% after 1-hour, 68 +/- 1.7% after 2 hours incubation at 4 degrees C. According to this, human peripheral T lymphocytes may be subdivided into at least three fractions on the basis of their affinity to SRBC. We called these fractions AFRC (no incubation), C1 RFC (1-hour incubation in cold after removing ARFC fraction) and C2RFC (2-hour incubation in cold after removing ARFC and C1RFC fractions). All the three fractions may be separated by repeated rosetting followed by Ficoll-Uropoculine gradient centrifugation. The purity and viability of T lymphocytes yielded from particular fractions was high. Effective yield of lymphocytes varied between the particular fractions ranging between 100% (for AFRC) and 30% (for C2RFC).