Establishment of a human B cell line that proliferates in response to B cell growth factor.
A human B cell line which shows a marked dose dependence on B cell growth factor (BCGF) when cultured in less than or equal to 2% serum has been established. Human B lymphocytes were obtained from peripheral blood of normal donors and cultured in the presence of anti-IgM (mu chain specific) and BCGF. Frequent refeedings with fresh medium containing BCGF and anti-IgM led to the establishment of a long term cultured human B cell line, HAB-40. Phenotyping of HAB-40 revealed that the cell population consisted predominantly of IgM-bearing (72%) and B1 (100%) positive cells. This B cell line consistently secreted IgM and IgG when co-cultured in the presence of PMA, anti-IgM and beta or gamma interferon (IFN). Also, it was Epstein-Barr virus nuclear antigen (EBNA) positive (100%). HAB-40 cells have been successfully maintained in the presence of BCGF without anti-IgM for over a year. Removal of BCGF led to the rapid loss of viable cells in cultures containing less than 2% serum. HAB-40 cells in microassays exhibited a marked dose-dependent incorporation of [3H]thymidine in response to BCGF in the absence of any exogenous stimulants such as anti-IgM or Staphylococcus aureus Cowan I (SAC). Recombinant interleukin 2 (IL-2) failed to augment the [3H]thymidine uptake by these B cells despite the low density expression of Tac antigen (IL-2 receptor) on their cell surface, or even when the cells were stimulated with phorbol myristate acetate (PMA) to express higher density of Tac antigen (48%). HAB-40 cells could be maintained in BCGF which was partially purified to deplete it of other contaminating proteins. None of the seven well established EBNA-positive human B cell lines nor two chronic B lymphocytic leukemia (B-CLL) cell lines that were tested showed BCGF dependence. The same BCGF-active chromatographic fractions that were active on HAB-40 cells also stimulated BCL1 and normal human B cells stimulated with anti-IgM. In the presence of less than or equal to 2% serum proteins this cell line provides a simple, reproducible assay for BCGF even in the presence of contaminant IL-2.
Abe
H
H
Rossio
J L
JL
Ruscetti
F W
FW
Matsushima
K
K
Oppenheim
J J
JJ
D000888:Antibodies, Anti-Idiotypic; D000954:Antigens, Surface / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens; D006133:Growth Substances / Q000494:pharmacology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D007372:Interferons / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D015847:Interleukin-4; D007945:Leukemia, Lymphoid / Q000473:pathology; D008222:Lymphokines / Q000494:pharmacology*; D008934:Mitogens / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7
D000888:Antibodies, Anti-Idiotypic; D000954:Antigens, Surface; D000956:Antigens, Viral; D019309:Epstein-Barr Virus Nuclear Antigens; D006133:Growth Substances; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007376:Interleukin-2; D008222:Lymphokines; D008934:Mitogens; D011994:Recombinant Proteins; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; C044203:anti-IgM; D015847:Interleukin-4; D007372:Interferons; D013755:Tetradecanoylphorbol Acetate