Identification of lymphocyte subpopulations in E-RFC-enriched and E-RFC-depleted cell fractions of fresh and cryopreserved lymphocytes.
Fresh lymphocytes and frozen-stored lymphocytes were separated into E-RFC-enriched and E-RFC-depleted cell fractions by density gradient centrifugation of sheep red blood cell (SRBC) rosette-forming cells (E-RFC), since the ability to form rosettes is primarily a T cell characteristic. Subpopulations of lymphocytes were identified, demonstrating the presence of cell surface markers: T cell specific antigens (T+), receptors for SRBC on T cells (E-RFC), Fc-receptors (FcR) for IgG type antibodies, and surface Ig (sIg). Our results indicate that, although the E-RFC-depleted fraction contains virtually no cells capable of binding SRBC, there is still a considerable proportion of T cells present in that fraction, as detected with the anti-T cell antiserum. Moreover, data are presented to indicate that the E-RFC-enriched fraction does not consist exclusively of T lymphocytes. Since this separation procedure is used frequently for the identification of the nature of effector cells in cell-mediated (CMC) and antibody-dependent cellular cytotoxicity (ADCC) assays, the identification of T cells in purified lymphocyte fractions by means of SRBC rosette formation may lead to a false conclusion as to the nature of the effector cells.
D001793:Blood Preservation; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D005615:Freezing; D006801:Humans; D007958:Leukocyte Count; D008214:Lymphocytes / Q000276:immunology*; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D012397:Rosette Formation; D013601:T-Lymphocytes / Q000276:immunology
D011947:Receptors, Antigen, B-Cell