Development and characterization of a human B cell line that responds to B cell growth factor but not interleukin 2

• Published in: The Journal of immunology (1950) Vol. 138; no. 8; pp. 2554 - 2560
• Main Authors: Tseng, CT, Springgate, CF, Piela, TH, Choi, YS
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Assoc Immnol 15.04.1987; American Association of Immunologists
• Subjects: Analysis of the immune response. Humoral and cellular immunity; Antigens, Surface - analysis; B-Lymphocytes - drug effects; B-Lymphocytes - immunology; Biological and medical sciences; Cell Division - drug effects; Cell Line; Cell Transformation, Viral; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Growth Substances - pharmacology; Herpesvirus 4, Human; Humans; Immunobiology; Immunoglobulin G - secretion; Interferon-gamma - pharmacology; Interleukin-2 - pharmacology; Interleukin-4; Lymphokines - pharmacology; Lymphokines, interleukins ( function, expression); Phenotype; Recombinant Proteins - pharmacology; Regulatory factors and their cellular receptors; Spleen - cytology; Human; Cell proliferation; B-cell growth factor; Membrane receptor; Cell line; Interleukin 2; B-Lymphocyte; Lymphokine; Lymphoblastoid cell; Growth factor
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.138.8.2554

The human lymphoblastoid cell line we present here proliferated in response to a 14,000 m.w. B cell growth factor (BCGF), and not to interleukin 2 (IL 2). This cell line, designated B-A3, was established by Epstein Barr virus (EBV) transformation of Staphylococcus aureus Cowan I (SAC)-activated spleen B cells, and has been maintained in RPMI 1640 medium complemented with 15% fetal calf serum (FCS) without the addition of other exogenous growth factors. A proliferative response, as measured by [3H]thymidine uptake of B-A3 cells was significantly induced by either commercial IL 2-free human BCGF preparations, or phytohemagglutinin-stimulated mixed lymphocyte culture supernatant at all FCS concentrations used in the assay. The most marked proliferation due to BCGF, however, was observed in the absence of FCS. This BCGF-induced proliferation was not influenced by IL 2 or interferon-gamma (IFN-gamma), because both recombinant IL 2 and IFN-gamma failed to induce proliferation. The response of B-A3 cells to a specific BCGF was additionally indicated by the responsiveness of this cell line to BCGF purified by a series of chromatographic steps. The BCGF to which B-A3 cells responded had a m.w. of 14,000 and was similar to low m.w. BCGF reported from other laboratories. Surface characterization of B-A3 cells, analyzed by flow cytometry with a panel of monoclonal antibodies, demonstrated that the majority of B-A3 cells were stained positively with Leu-12, HLA-DR, and surface IgG markers, whereas staining with surface IgM, IgD markers, pan T cell markers (Leu-4 and Leu-9), and IL 2 receptor (Tac) were consistently negative. Taken together, the human lymphoblastoid cell line we present here responded specifically to a low m.w. BCGF. This cell line may be of value in the purification of BCGF to homogeneity, in studies of the interactions of BCGF with human B cells, and in the identification of the BCGF receptor.