Phorbol myristate acetate modulates calcium ion-dependent superoxide anion generation induced by a monoclonal antibody raised against polymorphonuclear leukocytes.
We used a monoclonal antibody, YI 51, raised against human polymorphonuclear leukocytes (PMN) to induce superoxide anion (O2-) generation in cells. Although YI 51 alone played only a small part in inducing O2- generation in PMN, the amount of O2- generation induced in 5 X 10(5) PMN was 3.7 to 5.5 nmol/min when F(ab')2 fragments of rabbit anti-mouse immunoglobulin antibody were added as a cross-linking agent. This O2- -inducing activity was high compared with that of wheat germ agglutinin (WGA), insoluble immunoglobulin G immune complexes (IC), or phorbol myristate acetate (PMA). The binding of YI 51 and soluble immunoglobulin G IC to PMN was not reciprocally inhibitory, indicating that YI 51 does not interfere with ligand binding to the Fc receptor-binding site. In the absence of calcium ion (Ca2+), O2- generation induced by YI 51 decreased to 10 to 20% of that in the presence of Ca2+. In contrast, O2- generation in response to WGA, IC, or PMA under Ca2+-free conditions was not affected. When PMN were pretreated with low concentrations of PMA (10(-10) to 10(-9) M), the amount of O2- generation by the cells in response to YI 51 in Ca2+-free buffer was enhanced in a concentration-dependent manner. It also equaled the O2- generated by the cells in buffer containing Ca2+. In cells pretreated with PMA, the amount of O2- induced by WGA was enhanced two- to threefold over that in untreated cells. In contrast, there was no augmentation over untreated cells with stimulation by IC. These results suggest that YI 51, IC, and WGA induce O2- generation in human PMN in different manners.
Ichinose
Y
Y
Department of Chest Surgery, National Kyushu Cancer Center, Fukuoka, Japan.
Hara
N
N
Ohta
M
M
Motohiro
A
A
Kuda
T
T
Aso
H
H
Yagawa
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000502:physiology*; D000936:Antigen-Antibody Complex / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D001666:Binding Sites, Antibody / Q000187:drug effects; D002118:Calcium / Q000502:physiology*; D005110:Extracellular Space / Q000502:physiology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D011961:Receptors, Fc / Q000032:analysis / Q000378:metabolism; D017452:Receptors, IgG; D011975:Receptors, Mitogen / Q000187:drug effects; D013481:Superoxides / Q000096:biosynthesis*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000943:Antigens, Differentiation; D011961:Receptors, Fc; D017452:Receptors, IgG; D011975:Receptors, Mitogen; D014909:Wheat Germ Agglutinins; D013481:Superoxides; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium