Edited by Ruslan Medzhitov, Yale University School of Medicine, New Haven, CT, and approved January 24, 2017 (received for review September 1, 2016)
Understanding the mechanisms underlying the development of senescence and the consequences related to the accumulation of senescent cells is a major focus of ongoing research. Our report shows that senescent cells express a form of oxidized vimentin on their cell surface and that oxidized vimentin is secreted into the blood of senescence-prone senescence-accelerated mouse prone 8 mice. Given the growing evidence that oxidized proteins are involved in the development of human diseases, the detection and monitoring of secreted proteins like malondialdehyde-modified vimentin is certain to become a vital and noninvasive biomarker for studying senescence and monitoring age-related illnesses.
Studying the phenomenon of cellular senescence has been hindered by the lack of senescence-specific markers. As such, detection of proteins informally associated with senescence accompanies the use of senescence-associated β-galactosidase as a collection of semiselective markers to monitor the presence of senescent cells. To identify novel biomarkers of senescence, we immunized BALB/c mice with senescent mouse lung fibroblasts and screened for antibodies that recognized senescence-associated cell-surface antigens by FACS analysis and a newly developed cell-based ELISA. The majority of antibodies that we isolated, cloned, and sequenced belonged to the IgM isotype of the innate immune system. In-depth characterization of one of these monoclonal, polyreactive natural antibodies, the IgM clone 9H4, revealed its ability to recognize the intermediate filament vimentin. By using 9H4, we observed that senescent primary human fibroblasts express vimentin on their cell surface, and MS analysis revealed a posttranslational modification on cysteine 328 (C328) by the oxidative adduct malondialdehyde (MDA). Moreover, elevated levels of secreted MDA-modified vimentin were detected in the plasma of aged senescence-accelerated mouse prone 8 mice, which are known to have deregulated reactive oxygen species metabolism and accelerated aging. Based on these findings, we hypothesize that humoral innate immunity may recognize senescent cells by the presence of membrane-bound MDA-vimentin, presumably as part of a senescence eradication mechanism that may become impaired with age and result in senescent cell accumulation.
Author contributions: D.F., A.S.G., A.L.O., M.P.A., V.M., O.B.C., and A.V.G. designed research; D.F., C.M.R., S.A.-S., A.S.G., P.K., O.V.K., E.S., L.P.V., M.W., M.P.A., and V.M. performed research; D.F., C.M.R., S.A.-S., A.S.G., P.K., E.S., A.L.O., M.P.A., O.B.C., and A.V.G. analyzed data; and D.F., A.S.G., V.M., and A.V.G. wrote the paper.
Conflict of interest statement: O.B.C. and A.V.G. are cofounders and shareholders of Everon Biosciences, a biotech company that funded this work and owns related intellectual property.
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