Phagocytosis of serum- and IgG-opsonized zymosan particles induces apoptosis through superoxide but not nitric oxide in macrophage J774A.1.
Phagocytosis of serum- and IgG-opsonized zymosan (SOZ and IOZ, respectively) particles into J774A.1 macrophages induced apoptosis of the cells, accompanied by the expression of p21(WAF1), one of cyclin-dependent protein kinase (CDK) inhibitors. Furthermore, phagocytosis of SOZ and IOZ particles into macophages induced superoxide formation. Tat-superoxide dismutase (SOD), which is readily transduced into the cells using Tat-domain, protected the cells from the apoptosis induced by phagocytosis of SOZ and IOZ particles. lipopolysaccharide (LPS) /interferon-gamma (IFN-gamma) also caused the apoptosis of the cells. However, Tat-SOD could not protect the cells from LPS/IFN-gamma induced apoptosis, suggesting that apoptosis mechanisms involved are different from each other. In the present study, we determined the amounts of nitric oxide (NO) produced by SOZ, IOZ, and LPS/IFN-gamma, and found that SOZ and IOZ did not induce the generation of NO in macrophages, whereas LPS/ IFN-gamma did. The apoptosis due to phagocytosis was accompanied with the release of cytochrome c from mitochondrial membrane to cytosolic fraction. Furthermore, SOZ and IOZ induced the cleavage of procasapase-3 (35 kDa) to give rise to an active caspase-3 (20 kDa), which was blocked by Tat- SOD but not by 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), a scavenger of NO. On the other hand, LPS/IFN-gamma caused the activation of procaspase-3, which was blocked by PTIO but not by Tat-SOD. Taken together, phagocytosis of SOZ and IOZ particles induced apoptosis through superoxide but not NO in macrophages, accompanied with the release of cytochrome c and the activation of caspase-3.
Kim
Jun-Sub
JS
Department of Biochemistry, College of Medicine, Hallym University, Chunchon 200-702, Korea.
Kwon
Hyeok-Yil
HY
Choi
Won-Ho
WH
Jeon
Chan-Young
CY
Kim
Jong-Il
JI
Kim
Jaebong
J
Lee
Jae-Yong
JY
Kim
Yong-Sun
YS
Park
Jae-Bong
JB
D017209:Apoptosis / Q000276:immunology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis; D045304:Cytochromes c / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology*; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D009895:Opsonin Proteins / Q000276:immunology; D010587:Phagocytosis / Q000502:physiology*; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D015054:Zymosan
D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D007074:Immunoglobulin G; D008070:Lipopolysaccharides; D009895:Opsonin Proteins; D013481:Superoxides; D009569:Nitric Oxide; D007371:Interferon-gamma; D045304:Cytochromes c; D015054:Zymosan; D013482:Superoxide Dismutase; D053148:Caspase 3; D020169:Caspases