Catalase Plays a Critical Role in the CSF-independent Survival of Human Macrophages via Regulation of the Expression of BCL-2 Family

Abstract

A-M⌽, but not M-M⌽, can produce both extracellular catalase and cell-associated catalase in a CSF-independent manner. Intracellular glutathione levels were kept equivalent in these M⌽s, both in the presence or absence of CSF. These results indicate a critical role of extracellular catalase in the survival of human macrophages via regulation of the expression of BCL-2 family genes. Human tissue macrophages (M⌽)2 play important roles for homeostasis, and in vivo alveolar (A)-M⌽ acquire a strong antioxidant phenotype that contributes to prevention of the oxidant burst in an aerobic environment and can survive for long periods (1). In a previous study, we reported that human A-M⌽ and GM-CSF-induced monocyte-derived macrophages (GM-M⌽) are resistant to hydrogen peroxide (H 2 O 2 ) via their high basal and inducible levels of catalase activity and that M-CSF-induced monocyte-derived macrophages (M-M⌽) are sensitive to low levels of H 2 O 2 with low levels of catalase activity (2). GM-M⌽ is phenotypically identical to A-M⌽, whereas M-M⌽ closely resembles peritoneal M⌽ in respect to morphology, cell surface antigen expression, and several biological functions (2-7). In vivo A-M⌽ express BCL-2 family proteins such as BCL-2 and BCL-X L that prevent H 2 O 2 -induced apoptosis via inhibition of caspase-3 or -9 activation and cytochrome c release from mitochondria (8 -11). These findings suggest that a high level of catalase activity enables long survival of GM-M⌽ and A-M⌽ with positive regulation of BCL-2 family protein.In this study, we found that M-M⌽ absolutely require CSF for their survival and express high levels of BCL-2 gene and protein in the presence of M-CSF. In contrast to M-M⌽, GM-M⌽ and A-M⌽ can survive in the absence of CSF via high levels of BCL-X L gene and protein. We further examined the relation between catalase activity and distinct expression of BCL-2 family protein and make clear the roles of CSF in the regulation of catalase activity and BCL-2 family protein in human tissue macrophages under the influence of oxidative stress. EXPERIMENTAL PROCEDURESPreparation and Culture of Macrophages-Monocytes (Mo) were obtained from peripheral blood mononuclear cells of normal healthy volunteers using a magnetic cell separation system (MACS; Miltenyi Biotec, Bergisch Gladbach, Germany) with anti-CD14 monoclonal antibody-coated microbeads as described previously (7). CD14 ϩ Mo were cultured in RPMI 1640 medium (Nissui Seiyaku Co., Ltd., Tokyo, Japan), 3 mg/ml filtered glutamine (Sigma), 100 units/ml penicillin G potassium (Banyu Seiyaku Co., Ltd., Tokyo, Japan), 100 g/ml streptomycin (Meiji Seika Co., Ltd., Tokyo, Japan), 10% autoclaved NaHCO 3 , and 10% heat-inactivated fetal calf serum (Z. L. Bockneck Laboratories Inc., Ontario, Canada) with the following human recombinant cytokines at optimal concentrations: 5 ng/ml GM-CSF (Schering-Plough Japan, Osaka, Japan) or 50 ng/ml M-CSF (Morinaga Milk Industry Co., Ltd., Tokyo, Japan) at 37°C in humidified 5% CO 2 for 7 days. During the culture, Mo differentiated to M⌽. ...

Macrophage colony-stimulating factor is indispensable for repopulation and differentiation of Kupffer cells but not for splenic red pulp macrophages in osteopetrotic (op/op) mice after macrophage depletion

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Bronchiolar epithelial catalase is diminished in smokers with mild COPD

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Hypoxia Prolongs Monocyte/Macrophage Survival and Enhanced Glycolysis Is Associated with Their Maturation under Aerobic Conditions

“…Peripheral blood monocytes gradually die by apoptosis unless provided with a stimulus such as CSF-1 (42). We determined whether the prosurvival action of hypoxia on murine macrophage populations shown above extends to human monocytes.…”

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