Interleukin 7 Independent Development of Human B Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS 1996-09, Vol.93 (19), p.10348-10353
• Main Authors: Julie A. R. Pribyl, LeBien, Tucker W.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antibodies, Monoclonal; Antigens, CD - analysis; Antigens, CD19 - analysis; B lymphocytes; B-Lymphocytes - cytology; B-Lymphocytes - drug effects; B-Lymphocytes - immunology; Bone marrow; Bone Marrow - embryology; Bone Marrow - immunology; Bone marrow cells; Cell Differentiation; Cell growth; Cell lines; Cells, Cultured; Coculture Techniques; Cultured cells; Developmental biology; Fetus; Flow Cytometry; Goats; Hematopoiesis - immunology; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Humans; Immunity (Disease); Immunoglobulin kappa-Chains - immunology; Immunoglobulin lambda-Chains - immunology; Immunoglobulin mu-Chains - immunology; Immunophenotyping; Interleukin-7 - pharmacology; Kinetics; Mammals; Mice; Receptors, Immunologic - biosynthesis; Species Specificity; Stem cells; Stromal Cells; Time Factors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.19.10348

Mammalian hematopoietic stem cell (HSC) commitment and differentiation into lymphoid lineage cells proceed through a series of developmentally restricted progenitor compartments. A complete understanding of this process, and how it differs from HSC commitment and differentiation into cells of the myeloid/erythroid lineages, requires the development of model systems that support HSC commitment to the lymphoid lineages. We now describe a human bone marrow stromal cell culture that preferentially supports commitment and differentiation of human HSC to CD19$^{+}$ B-lineage cells. Fluorescence activated cell sorterpurified CD34$^{++}$/lineage$^{-}$ cells were isolated from fetal bone marrow and cultured on human fetal bone marrow stromal cells in serum-free conditions containing no exogenous cytokines. Over a period of 3 weeks, CD34$^{++}$/lineage$^{-}$ cells underwent commitment, differentiation, and expansion into the B lineage. Progressive changes included: loss of CD34, acquisition of and graded increases in the level of cell surface CD19, and appearance of immature B cells expressing $\mu $/$\kappa $ or $\mu $/$\lambda $ cell surface Ig receptors. The tempo and phenotype of B-cell development was not influenced by the addition of IL-7 (10 ng/ml), or by the addition of goat anti-IL-7 neutralizing antibody. These results indicate a profound difference between mouse and human in the requirement for IL-7 in normal B-cell development, and provide an experimental system to identify and characterize human bone marrow stromal cell-derived molecules crucial for human B lymphopoiesis.