title: Surfactant protein-A modulates LPS-induced TLR4 localization and signaling via β-arrestin 2.
abstract: The soluble C-type lectin surfactant protein (SP)-A mediates lung immune responses partially via its direct effects on alveolar macrophages (AM), the main resident leukocytes exposed to antigens. SP-A modulates the AM threshold of lipopolysaccharide (LPS) activity towards an anti-inflammatory phenotype both in vitro and in vivo through various mechanisms. LPS responses are tightly regulated via distinct pathways including subcellular TLR4 localization and thus ligand sensing. The cytosolic scaffold and signaling protein β-arrestin 2 acts as negative regulator of LPS-induced TLR4 activation. Here we show that SP-A neither increases TLR4 abundancy nor co-localizes with TLR4 in primary AM. SP-A significantly reduces the LPS-induced co-localization of TLR4 with the early endosome antigen (EEA) 1 by promoting the co-localization of TLR4 with the post-Golgi compartment marker Vti1b in freshly isolated AM from rats and wild-type (WT) mice, but not in β-arrestin 2(-/-) AM. Compared to WT mice pulmonary LPS-induced TNF-α release in β-arrestin 2(-/-) mice is accelerated and enhanced and exogenous SP-A fails to inhibit both lung LPS-induced TNF-α release and TLR4/EEA1 positioning. SP-A, but not LPS, enhances β-arrestin 2 protein expression in a time-dependent manner in primary rat AM. The constitutive expression of β-arrestin 2 in AM from SP-A(-/-) mice is significantly reduced compared to SP-A(+/+) mice and is rescued by SP-A. Prolonged endosome retention of LPS-induced TLR4 in AM from SP-A(-/-) mice is restored by exogenous SP-A, and is antagonized by β-arrestin 2 blocking peptides. LPS induces β-arrestin 2/TLR4 association in primary AM which is further enhanced by SP-A. The data demonstrate that SP-A modulates LPS-induced TLR4 trafficking and signaling in vitro and in vivo engaging β-arrestin 2.
authors.affiliation: Division of Cellular Pneumology, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.
authors.forename: Vicky
authors.initials: V
authors.lastname: Sender
authors.forename: Linda
authors.initials: L
authors.lastname: Lang
authors.forename: Cordula
authors.initials: C
authors.lastname: Stamme
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism*; D055786:Gene Knockout Techniques; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology*; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D037662:Pulmonary Surfactant-Associated Protein A / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D033921:Vesicular Transport Proteins / Q000378:metabolism; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
chemical_list: C000605258:Arrb2 protein, mouse; C000605257:Arrb2 protein, rat; D019390:Arrestins; D008070:Lipopolysaccharides; D037662:Pulmonary Surfactant-Associated Protein A; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D033921:Vesicular Transport Proteins; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; C093846:early endosome antigen 1
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