B-lineage colonies from normal, human bone marrow are initiated by B cells and their progenitors.
We have recently described a reproducible method whereby colonies containing cells that secrete immunoglobulin (Ig) can be grown from normal, human, adult bone marrow samples. The present report characterizes the cells that initiate these colonies. It is shown that all clonogenic cells express the CD19 surface antigen, as removal of these cells before plating in the B-cell colony assay abolished the subsequent growth of plaque-forming, B-lineage colonies. Cells from both the CD10+ and CD20+ B-lineage subpopulations initiated the growth of B-cell colonies, as removal of either subset resulted in a 50% reduction in the number of resulting B-cell colonies. The removal of activated B cells (CD23+), plasma cells (PCA-1+), or myeloid cells (CD13+) did not lead to a significant depletion in B-cell colony formation. Pre-B cells that were not yet committed to Ig light chain expression were also able to differentiate and proliferate into Ig-secreting colonies under the culture conditions used. Colonies initiated by these light chain uncommitted cells were distinguished using a replicate protein immunoblotting technique, which detects the simultaneous secretion of Ig kappa and Ig lambda from single colonies. These experiments provide evidence that the CD10 antigen is expressed on B-lineage cells before Ig light chain commitment, whereas CD20 is not. In conclusion, this B-cell colony assay provides a system for studying the differentiation of bone marrow-derived B cells and their precursors into Ig-secreting cells.
McGinnes
K
K
Division of Cell and Molecular Biology, Ontario Cancer Institute, Toronto, Canada.
Letarte
M
M
Paige
C J
CJ
D000911:Antibodies, Monoclonal / Q000276:immunology; D000921:Antibody-Producing Cells / Q000166:cytology; D015703:Antigens, CD / Q000378:metabolism; D018941:Antigens, CD19; D018951:Antigens, CD20; D000943:Antigens, Differentiation / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology*; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis; D007146:Immunoglobulin lambda-Chains / Q000096:biosynthesis; D066298:In Vitro Techniques; D015260:Neprilysin
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018941:Antigens, CD19; D018951:Antigens, CD20; D000943:Antigens, Differentiation; D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D015260:Neprilysin