p16(Ink4a) and senescence-associated β-galactosidase can be induced in macrophages as part of a reversible response to physiological stimuli

Constitutive p16(Ink4a) expression, along with senescence-associated beta-galactosidase (SA beta G), are commonly accepted biomarkers of senescent cells (SCs). Recent reports attributed improvement of the healthspan of aged mice following p16(Ink4a)-positive cell killing to the eradication of accumulated SCs. However, detection of p16(Ink4a)/SA beta G-positive macrophages in the adipose tissue of old mice and in the peritoneal cavity of young animals following injection of alginate-encapsulated SCs has raised concerns about the exclusivity of these markers for SCs. Here we report that expression of p16(Ink4a) and SA beta G in macrophages is acquired as part of a physiological response to immune stimuli rather than through senescence, consistent with reports that p16(Ink4a) plays a role in macrophage polarization and response. Unlike SCs, p16(Ink4a)/SA beta G-positive macrophages can be induced in p53-null mice. Macrophages, but not mesenchymal SCs, lose both markers in response to M1-[LPS, IFN-alpha, Poly(I:C)] and increase their expression in response to M2-inducing stimuli (IL-4, IL-13). Moreover, interferon-inducing agent Poly(I:C) dramatically reduced p16(Ink4a) expression in vivo in our alginate bead model and in the adipose tissue of aged mice. These observations suggest that the antiaging effects following eradication of p16(Ink4a)-positive cells may not be solely attributed to SCs but also to non-senescent p16(Ink4a)/SA beta G-positive macrophages.