Isolation of Human Mononuclear Cell Subsets by Counterflow Centrifugal Elutriation (CCE). II. Functional Properties of B-Lymphocyte-, T-Lymphocyte-, and Monocyte-Enriched Fractions

摘要

Counterflow centrifugal elutriation (CCE)), a technique which separates cells by size and density, was used to seperate human peripheral blood mononuclear cells into fractions enriched for T lymphocytes, B lymphocytes. These morphologically and phenotypically distinct fractions were analyzed for their ability to respond in several functional assays. B-Cell-enriched fractions devoid of monocytes did not proliferate nor produce significant quantities of lymphokines in response to antigens. These B cells did not proliferate to anti-IgM antibodies but not to anti-IgD antibodies. B-Cell fractions served as simulators of the autologous mixed-lymphocyte reaction (AMLR). T-Lymphocyte fractions were unable to respond to antigen challenge, but both proliferation and lymphokine production could be restored by the addition of monocytes. Monocyte fractions produced Prostaglandin E2 (PGE2) displayed chemotaxis, and functioned as stimulators in the AMLR. Thus, CCE appears to be a useful technique for reproducibly obtaining highly enriched subsets of human peripheral blood mononuclear cells with unique phenotypic and functional properties. These isolated populations can consequently be used to identify the independent and collaborative roles of the cells in immunological events.