Chonnam Med J.  2020 Jan;56(1):12-19. 10.4068/cmj.2020.56.1.12.

Differential Activation of Macrophages Based on Their Environment in Advanced Age

• Lee DY
• Lim JS
• Cho KA

• KMID: 2468141
• DOI: http://doi.org/10.4068/cmj.2020.56.1.12

Abstract

The macrophage displays functional and phenotypic diversity, which appears, in no small part, to stem from the ability of macrophages to adapt functionally to changes in their tissue microenvironment. Here, we describe the differential activity of peritoneal macrophages with or without the presence of thioglycollate (TG), an inflammatory drug that encouraged the recruitment of macrophages, during aging. The peritoneal-resident macrophages dramatically reduced in phagocytosis and pro-inflammatory cytokines secretion with aging, whereas the functions of macrophages recruited by TG were not significantly changed with aging. These results suggest that macrophages may be changed by their environment in advanced age, and could provide possible explanations for the controversial results regarding differential changes in macrophages in other papers.

Keyword

Macrophages; Thioglycollate; Inflammation; Environment

MeSH Terms

Aging
Cytokines
Inflammation
Macrophages*
Macrophages, Peritoneal
Phagocytosis
Cytokines

Figure

• 

  FIG. 1 Differential activation between tissue-resident and TG-elicited peritoneal macrophages from young mice. Peritoneum-resident (Mϕ)- and TG-elicited peritoneum macrophages (TG-Mϕ) were isolated from young mice (Y, 8 weeks old). Total cell numbers (n=5/each group) were counted (A), and phagocytotic activity (n=5/each group) was quantified (B). The ratio for Mϕ was arbitrarily defined as 100%, and the ratios for TG-Mϕ were expressed as relative values. (C) The cell surface expression of CD11b (M1 polarization marker) and CD206 (M2 polarization marker) in a F4/80-gated population of peritoneum-resident and TG-elicited peritoneum macrophages (n=5/each group) was analyzed by flow cytometry. (D) Cells (n=5/each group) were stimulated with LPS (5 mg/mL). IL-6, TNF-α, and IL-1β, and IL-10 levels were analyzed by ELISA. *p<0.05; **p<0.01. #p<0.05; ##p<0.01 compared with non-treated and LPS-treated macrophages. LPS: lipopolysaccharide, TG: thioglycollate.

• 

  FIG. 2 Differential activation between tissue-resident and TG-elicited peritoneal macrophages from old mice. Mϕ and TG-Mϕ were isolated from old mice (O, 24 months). Total cell numbers (n=5/each group) were counted (A), and phagocytotic activity (n=5/each group) was quantified (B). (C) The cell surface expression of CD11b and CD206 in a F4/80-gated population of Mϕ and TG-Mϕ (n=5/each group) was analyzed by flow cytometry. (D) Cells (n=5/each group) were stimulated with LPS (5 µg/mL). IL-6, TNF-α, IL-1β, IL-10 levels were analyzed by ELISA. *p<0.05; **p<0.01; ***p<0.001. #p<0.05 compared with non-treated and LPS-treated macrophages. NS indicates not significant.

• 

  FIG. 3 Comparison of functional alterations in peritoneum-resident macrophages from young and old mice. After isolation of Mϕ and TG-Mϕ from Y- and O-mice, total cell numbers (n=5/each group) were counted (A), and phagocytotic activity (n=5/each group) was quantified (B). **p<0.01; ***p<0.001. NS indicates not significant.

• 

  FIG. 4 Age-associated functional alterations of peritoneum-resident macrophages. After isolation of Mϕ and TG-Mϕ from Y- and O-mice, cells (n=5/each group) were stimulated with Salmonella extract proteins (Sal-P) (10 µg/mL), LPS (5 µg/mL), and Vv-FlaB (100 ng/mL). IL-6 (A), TNF-α (B), and IL-1β (C) levels were analyzed by ELISA. *p<0.05; NS indicates not significant. Vv-FlaB: Vibrio vulnificus major flagellin recombinant proteins.

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