title: S100A9 induced inflammatory responses are mediated by distinct damage associated molecular patterns (DAMP) receptors in vitro and in vivo.
abstract: Release of endogenous damage associated molecular patterns (DAMPs), including members of the S100 family, are associated with infection, cellular stress, tissue damage and cancer. The extracellular functions of this family of calcium binding proteins, particularly S100A8, S100A9 and S100A12, are being delineated. They appear to mediate their functions via receptor for advanced glycation endproducts (RAGE) or TLR4, but there remains considerable uncertainty over the relative physiological roles of these DAMPs and their pattern recognition receptors. In this study, we surveyed the capacity of S100 proteins to induce proinflammatory cytokines and cell migration, and the contribution RAGE and TLR4 to mediate these responses in vitro. Using adenoviral delivery of murine S100A9, we also examined the potential for S100A9 homodimers to trigger lung inflammation in vivo. S100A8, S100A9 and S100A12, but not the S100A8/A9 heterodimer, induced modest levels of TLR4-mediated cytokine production from human PBMC. In contrast, for most S100s including S100A9, RAGE blockade inhibited S100-mediated cell migration of THP1 cells and major leukocyte populations, whereas TLR4-blockade had no effect. Intranasal administration of murine S100A9 adenovirus induced a specific, time-dependent predominately macrophage infiltration that coincided with elevated S100A9 levels and proinflammatory cytokines in the BAL fluid. Inflammatory cytokines were markedly ablated in the TLR4-defective mice, but unexpectedly the loss of TLR4 signaling or RAGE-deficiency did not appreciably impact the S100A9-mediated lung pathology or the inflammatory cell infiltrate in the alveolar space. These data demonstrate that physiological levels of S100A9 homodimers can trigger an inflammatory response in vivo, and despite the capacity of RAGE and TLR4 blockade to inhibit responses in vitro, the response is predominately independent of both these receptors.
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Allison L
authors.initials: AL
authors.lastname: Miller
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Marlon
authors.initials: M
authors.lastname: Rebelatto
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Yambasu
authors.initials: Y
authors.lastname: Brewah
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Rowe
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Lori
authors.initials: L
authors.lastname: Clarke
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Meggan
authors.initials: M
authors.lastname: Czapiga
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Kim
authors.initials: K
authors.lastname: Rosenthal
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America; Laboratory of Human Retrovirology, Applied and Developmental Directorate, Building 550 Room 126, Leidos Biomedical Research Inc, Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702, United States of America.
authors.forename: Tomozumi
authors.initials: T
authors.lastname: Imamichi
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Chew-Shun
authors.initials: CS
authors.lastname: Chang
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Partha S
authors.initials: PS
authors.lastname: Chowdhury
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Brian
authors.initials: B
authors.lastname: Naiman
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Laboratory of Molecular Immunoregulation, National Cancer Institute, Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702, United States of America.
authors.forename: De
authors.initials: D
authors.lastname: Yang
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Alison A
authors.initials: AA
authors.lastname: Humbles
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Ronald
authors.initials: R
authors.lastname: Herbst
authors.affiliation: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States of America.
authors.forename: Gary P
authors.initials: GP
authors.lastname: Sims
mesh_terms: D000818:Animals; D040502:Calgranulin B / Q000494:pharmacology*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D000067759:Receptor for Advanced Glycation End Products / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051197:Toll-Like Receptor 4 / Q000378:metabolism
chemical_list: D040502:Calgranulin B; D000067759:Receptor for Advanced Glycation End Products; C576329:S100A9 protein, mouse; D051197:Toll-Like Receptor 4