Antigenic Phenotype and Functional Characterization of Human Tonsil B Cells

• Published in: Blood 1988-04, Vol.71 (4), p.1048-1055
• Main Authors: Gadol, Nancy, Peacock, Margaret A., Ault, Kenneth A.
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal; Antigens, Differentiation, B-Lymphocyte - analysis; Antigens, Differentiation, B-Lymphocyte - immunology; Antigens, Surface - analysis; Antigens, Surface - immunology; B-Lymphocytes - classification; B-Lymphocytes - immunology; B-Lymphocytes - physiology; Biological and medical sciences; Cell Separation; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Fundamental immunology; HLA-D Antigens - analysis; Humans; Immunobiology; Immunohistochemistry; Lymphocyte Function-Associated Antigen-1; Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation; Palatine Tonsil - analysis; Palatine Tonsil - anatomy & histology; Palatine Tonsil - cytology; Phenotype; Receptors, Antigen, B-Cell - analysis; Staining and Labeling; Suspensions
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V71.4.1048.1048

Human tonsil cells were labeled with anti-leu-16, a mono-clonal antibody (MoAb) that recognizes the CD20 antigen and that is specific for B cells. Two populations of B cells were identified by flow cytometry on the basis of antigen density. One labeled brightly with anti-Leu-16, and the other labeled at a level comparable to blood B cells. These two populations were characterized with a panel of MoAbs in two- and three-color flow cytometric studies and appeared to correspond to germinal-center and mantle-zone B cells. The pattern of staining of anti-Leu-16 on sections of frozen tonsil supported this characterization. Anti-Leu-16 labeled germinal center cells more intensely than mantle zone cells and stained a few scattered B cells in the interfollicular zone. The ability of each Leu-16+ population to secrete IgG and IgM in response to mitogens was measured in a particle immunofluorescence assay. Dim Leu-16+ B cells (small, resting B cells and a subpopulation of preactivated cells) secreted IgG and IgM in response to pokeweed mitogen (PWM) but only IgG in response to B cell growth factor (BCGF). Bright Leu-16+ B cells (small to large activated cells and possibly memory cells) did not respond to PWM but secreted IgG in response to BCGF. The functional responses of dim Leu-16+ and bright Leu-16+ B cells were consistent with their identification as mantle-zone and germinal-center B cells. Phenotypic identification and functional studies of mantle-zone and germi-nal-center B cells may help clarify the differentiation pathway within the germinal center.© 1988 by Grune & Stratton, Inc. 0006-4971/88/7104-0018$ 3.00/0