Isolation of monocyte-depleted and monocyte-rich fractions from human mononuclear cells.
Utilizing a recently reported characteristic of monocytes to aggregate in cold, a new procedure for obtaining monocyte-rich and monocyte-depleted mononuclear fractions from human blood is described. After aggregation in cold and adherence to plastic, a fraction containing 88% monocyte (LeuM3 CD14+) is obtained. After treating the supernatant, which separates from the aggregates, with a monocyte lysosomotropic agent (L-Leucine Methyl Ester), a fraction containing 0% monocyte (LeuM3 CD14+) and 88% lymphocytes (Leu4 CD3+ and Leu 12 CD19+) is obtained. In round-bottom culture wells, this monocyte-depleted fraction produced immunoglobulins in response to pokeweed mitogen (5,826 +/- 2,356 IgM micrograms/ml), to a degree not significantly different that that produced in the presence of monocytes (7,426 +/- 3,347 IgM micrograms/ml). This suggests the presence of cells in the lymphocyte fraction, that are not monocytes but are capable of antigen presenting.
Rubinstien
E
E
Department of Medicine, Saint Francis Hospital and Medical Center, Hartford, Connecticut 06105.
Ballow
M
M
D000328:Adult; D000917:Antibody Formation; D000938:Antigen-Presenting Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003080:Cold Temperature; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000502:physiology; D011043:Pokeweed Mitogens / Q000494:pharmacology
D011043:Pokeweed Mitogens; C030365:leucine methyl ester; D007930:Leucine