Identification of histamine-production-increasing factor produced by stimulated RBL-2H3 rat basophilic leukemia cells as granulocyte-macrophage colony-stimulating factor.
When RBL-2H3 rat basophilic leukemia cells were stimulated by antigen or the Ca2+ ionophore A23187, the activity to increase histamine production by rat bone marrow cells in the conditioned medium increased time-dependently. To characterize the histamine-production-increasing factor (HPIF) produced by RBL-2H3 cells, the conditioned medium was collected 8 h after stimulation by A23187, and the factor was purified by three-step chromatography, the specific activity being increased by 9000-fold. The partial amino acid sequence of the peptide obtained by S. aureus V8 protease digestion was identical to the internal amino acid sequence of rat granulocyte-macrophage colony-stimulating factor (GM-CSF). In addition, GM-CSF mRNA levels in RBL-2H3 cells were increased by A23187 with a peak at 4 h. Furthermore, recombinant rat GM-CSF increased histamine production by rat bone marrow cells. These findings suggested that HPIF produced by the stimulated RBL-2H3 cells is GM-CSF. Possible significant roles of HPIF at the late phase of allergic inflammation are discussed.
Department of Pathophysiological Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Aoba Aramaki, Aoba-ku, Sendai, Miyagi 980-8578, Japan.
M
M
Shiraishi
N
N
Hirasawa
S
S
Mue
K
K
Ohuchi
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000276:immunology; D001854:Bone Marrow Cells / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D017077:Culture Media, Conditioned; D004140:Dinitrophenols / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006632:Histamine / Q000096:biosynthesis*; D007476:Ionophores / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000276:immunology; D014407:Tumor Cells, Cultured
D000941:Antigens; D017077:Culture Media, Conditioned; D004140:Dinitrophenols; D007476:Ionophores; D012333:RNA, Messenger; D011994:Recombinant Proteins; C034638:dinitrophenyl-bovine serum albumin; D012710:Serum Albumin, Bovine; D000001:Calcimycin; D006632:Histamine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor