Establishment of murine macrophage-like mutant and hybrid cell lines: comparative analysis of the differentiation induced by 1 alpha,25-dihydroxyvitamin D3 and recombinant murine interferon-gamma.
The purpose of this study is to establish the monocyte/macrophage-like cell lines which are sensitive to potent systemic and local factors, 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2 VD3) and interferon-gamma (IFN-gamma). We established two variant mouse macrophage-like cell lines, whose responses to 1 alpha,25(OH)2 VD3 and IFN-gamma differed from one another. The AH-sensitive mutant cell line (G3) was induced by allowing P388D1 tolerant to 8-azaguanine. G3 mutant cells were then fused with the 1 alpha,25(OH)2 VD3-stimulated bone marrow cells isolated from DBA/2 mice. After AH selection the hybrid cell line (XC) was established. The G3 mutant cell line and the XC hybrid cell line had macrophage-like characteristics, such as surface antigens, Fc receptor, C3 receptor, and lysosomal enzymes. The treatment of G3 mutant cells with 1 alpha,25(OH)2 VD3 inhibited cell proliferation with morphological changes, and increased acid phosphatase activity, phagocytic activity, and F4/80 antigen expression on the cell surface. In contrast, IFN-gamma inhibited cell proliferation without effect on acid phosphatase activity and phagocytic activity but increased F4/80 antigen expression. In XC hybrid cells, on the other hand, IFN-gamma, but not 1 alpha,25(OH)2 VD3, inhibited cell proliferation with morphological changes but increased phagocytic activity and F4/80 antigen expression. In addition, IFN-gamma, but not 1 alpha,25(OH)2 VD3, dose-dependently increased multinucleated cell formation of both cells. These findings suggest that the G3 mutant cell line with macrophage-like characteristics is 1 alpha,25(OH)2 VD3- and IFN-gamma-sensitive, and that the XC hybrid cell line is, despite its macrophage-like characteristics, only IFN-gamma-sensitive. Therefore, these newly established cell lines will provide useful systems in studying the differentiation of monocyte/macrophage lineage.
Orikasa
M
M
Department of Pharmacology, Niigata University School of Dentistry, Japan.
Kawase
T
T
Shimizu
F
F
Suzuki
A
A
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D006822:Hybrid Cells / Q000187:drug effects; D007371:Interferon-gamma / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D009154:Mutation; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D007371:Interferon-gamma; D000135:Acid Phosphatase; D002117:Calcitriol