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Differential stimulation requirements for IL-12 production among clones of macrophage hybridomas
M Taneichi1, A Horino, S Naito
1Department of Bacterial and Blood Products, National Institute of Infectious Diseases, Tokyo, Japan.
We have previously established cloned macrophage hybridomas by somatic cell fusion of the macrophage tumor P388D1 of DBA/2 (H-2d) origin with splenic adherent cells of CKB mice (H-2k). Several cloned lines displayed the serologic and functional characteristics of macrophages. In this study, we evaluated the ability of these hybridomas to produce IL-12 after combined stimulation with IFN-gamma and lipopolysaccharide (LPS). The patterns of IL-12 production by these cloned macrophages fell into three groups. The first group produced IL-12 on stimulation with LPS in combination with IFN-gamma pretreatment, the second group produced IL-12 on stimulation with LPS regardless of the pretreatment with IFN-gamma, and the third group did not produce IL-12 at all on stimulation with IFN-gamma and LPS. None of the macrophage clones tested produced IL-12 constitutively. The results correlated well with IL-12 p40 mRNA expression in those macrophages as detected by RT-PCR. These results suggest the differential stimulation requirements for IL-12 production among macrophages at a clonal level.
We have previously established cloned macrophage hybridomas by somatic cell fusion of the macrophage tumor P388D1 of DBA/2 (H-2d) origin with splenic adherent cells of CKB mice (H-2k). Several cloned lines displayed the serologic and functional characteristics of macrophages. In this study, we evaluated the ability of these hybridomas to produce IL-12 after combined stimulation with IFN-gamma and lipopolysaccharide (LPS). The patterns of IL-12 production by these cloned macrophages fell into three groups. The first group produced IL-12 on stimulation with LPS in combination with IFN-gamma pretreatment, the second group produced IL-12 on stimulation with LPS regardless of the pretreatment with IFN-gamma, and the third group did not produce IL-12 at all on stimulation with IFN-gamma and LPS. None of the macrophage clones tested produced IL-12 constitutively. The results correlated well with IL-12 p40 mRNA expression in those macrophages as detected by RT-PCR. These results suggest the differential stimulation requirements for IL-12 production among macrophages at a clonal level.