Immunoglobulin-positive mononuclear cells in human peripheral blood: detection by mixed anti-globulin and direct anti-globulin-rosetting reactions.
Ig-bearing mononuclear cells were identified in Ficoll-Hypaque preparations of human peripheral blood by using mixed anti-globulin (MAG) and direct anti-globulin rosettes; indicator cells consisted of sheep erythrocytes coated with human F(ab')2 or anti-F(ab')2 antibody, respectively. Of the cell population isolated from 10 normal subjects, a mean of 68% was lymphocytes. However, fewer than 50% of the cells with detectable surface Ig were lymphocytes. On viable cell preparations using chromic chloride-treated sheep erythrocytes (CrCl3SRBC) coated with anti-F(ab')2 antibody, a mean of 20.1% of the lymphocytes formed rosettes, i.e., were B. Up to 6% of peripheral blood lymphocytes formed mixed Ig-rosettes and E-rosettes. On viable lymphocytes using F(ab')2-coated CrCl3SRBC, MAG rosettes were insensitive in detection of B lymphocytes. Formaldehyde treatment of lymphocytes increased the number of B cells detectable to 25.5% of the lymphocyte population. Study of T-enriched and B-enriched populations showed that the observed increase in B cell reactivity was real and not due to MAG-rosetting T cells. A one-stage procedure for T and B lymphocyte separation is described.
D000888:Antibodies, Anti-Idiotypic; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D004912:Erythrocytes / Q000276:immunology; D005557:Formaldehyde / Q000494:pharmacology; D006386:Hemagglutination Tests; D006801:Humans; D007104:Immune Adherence Reaction; D008214:Lymphocytes / Q000276:immunology*; D009000:Monocytes / Q000276:immunology*; D011947:Receptors, Antigen, B-Cell / Q000032:analysis*; D013601:T-Lymphocytes / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D011947:Receptors, Antigen, B-Cell; D005557:Formaldehyde