title: Senescent cells expose and secrete an oxidized form of membrane-bound vimentin as revealed by a natural polyreactive antibody.
abstract: 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.
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263; dfrescas@tartiscorp.com ochernova@tartiscorp.com andrei.gudkov@roswellpark.org.
authors.forename: David
authors.initials: D
authors.lastname: Frescas
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Christelle M
authors.initials: CM
authors.lastname: Roux
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Semra
authors.initials: S
authors.lastname: Aygun-Sunar
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Anatoli S
authors.initials: AS
authors.lastname: Gleiberman
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Peter
authors.initials: P
authors.lastname: Krasnov
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037.
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Kurnasov
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Evguenia
authors.initials: E
authors.lastname: Strom
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Lauren P
authors.initials: LP
authors.lastname: Virtuoso
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Michelle
authors.initials: M
authors.lastname: Wrobel
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037.
authors.forename: Andrei L
authors.initials: AL
authors.lastname: Osterman
authors.affiliation: Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY 14263.
authors.forename: Marina P
authors.initials: MP
authors.lastname: Antoch
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263.
authors.forename: Vadim
authors.initials: V
authors.lastname: Mett
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263; dfrescas@tartiscorp.com ochernova@tartiscorp.com andrei.gudkov@roswellpark.org.
authors.forename: Olga B
authors.initials: OB
authors.lastname: Chernova
authors.affiliation: Everon Biosciences, Inc., Buffalo, NY 14263; dfrescas@tartiscorp.com ochernova@tartiscorp.com andrei.gudkov@roswellpark.org.|Department of Cell Stress Biology, Roswell Park Cancer Institute, Buffalo, NY 14263.
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Gudkov
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D056724:Immunity, Humoral / Q000502:physiology; D007113:Immunity, Innate / Q000502:physiology; D007075:Immunoglobulin M / Q000378:metabolism; D007382:Intermediate Filaments / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D017382:Reactive Oxygen Species / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D000906:Antibodies; D015415:Biomarkers; D007075:Immunoglobulin M; D017382:Reactive Oxygen Species; D014746:Vimentin; D008315:Malondialdehyde; D001616:beta-Galactosidase
keywords: SAMP8; aging; biomarker; malondialdehyde; oxidative posttranslational modifications