Delineation and characterization of human B cell subpopulations at various stages of activation by using a B cell-specific monoclonal antibody.
Human tonsillar B cells were separated into three distinct subpopulations, Ba-/IgD+, Ba+/IgD+, and Ba+/IgD-, by using a B cell-specific monoclonal antibody (anti-Ba) that recognizes only activated B cells, and anti-IgD antibody. Stimulation of Ba-/IgD+ cells with anti-mu plus PHA-conditioned culture supernatant (PHA-sup) or TPA induced Ba+/IgD+ cells, which reverted to Ba-/IgD+ phenotype in the absence of continuous stimulation. Further stimulation of Ba+/IgD+ cells with several B cell activators, such as TPA plus anti-mu or PWM plus T cells, resulted in the loss of IgD expression. Three-color FACS analysis showed that the expression of transferrin receptor (TFR) was at its maximum in Ba+/IgD- cells, and the intensity of this expression was proportional to that of Ba expression in Ba+/IgD+ cells. PHA-sup induced maximum proliferation in Ba+/IgD- cells, and the degree of response was a function of the intensity of Ba expression in Ba+/IgD+ cells. PHA-sup or purified BCDF (BSF-2) induced Ig secretion preferentially in Ba+/IgD- cells. Taken together, these results show that resting B cells (Ba-/IgD+) are activated into Ba+/IgD+ cells, and then into Ba+/IgD- cells, under mitogenic stimulation, and BCDF induces the final maturation of Ba+/IgD- cells into Ig-secreting cells. Ba+/IgD- cells, which maximally expressed TFR as well as Ba and displayed maximum proliferative response to PHA-sup, did not express any Tac antigen. On the other hand, in vitro activated B cells expressed Ba and TFR as well as Tac antigen.
H
H
Kikutani
H
H
Nakamura
R
R
Sato
R
R
Kimura
K
K
Yamasaki
R R
RR
Hardy
T
T
Kishimoto
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D001402:B-Lymphocytes / Q000145:classification* / Q000276:immunology; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006133:Growth Substances / Q000032:analysis / Q000276:immunology; D006801:Humans; D007072:Immunoglobulin D / Q000276:immunology; D007136:Immunoglobulins / Q000096:biosynthesis; D015847:Interleukin-4; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000032:analysis / Q000276:immunology; D010641:Phenotype; D011947:Receptors, Antigen, B-Cell / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D000954:Antigens, Surface; D000939:Epitopes; D006133:Growth Substances; D007072:Immunoglobulin D; D007136:Immunoglobulins; D008222:Lymphokines; D011947:Receptors, Antigen, B-Cell; D015847:Interleukin-4