We recently provided evidence that anti-inflammatory macrophage activation, i.e., the inhibition of constitutive and signal-induced NF-κB activity by the pulmonary collectin surfactant protein (SP)-A, critically involves a promoted stabilization of IκB-α, the predominant inhibitor of NF-κB, via posttranscriptional mechanisms comprising the activation of atypical (a)PKCζ. SP-A uptake and degradation by alveolar macrophages (AMφ) occur in a receptor-mediated, clathrin-dependent manner. However, a mutual link between endocytosis of and signaling by SP-A remains elusive. The aim of this study was to investigate whether clathrin-mediated endocytosis (CME) of SP-A by AM is a prerequisite for its modulation of the IκB-α/NF- κB pathway. The inhibition of clathrin-coated pit (CCP) formation and clathrin-coated vesicle (CCV) formation/budding abrogates SP-A-mediated IκB-α stabilization and SP-A-mediated inhibition of LPS-induced NF-κB activation in freshly isolated rat AMφ, as determined by Western analysis, fluorescence-activated cell sorting, confocal microscopy, and EMSA. Actin depolymerization and inhibition of CCP formation further abolished SP-A-mediated inhibition of LPS-induced TNF-α release, as determined by ELISA. In addition, SP-A-induced atypical PKCζ activation was abolished by pretreatment of AMφ with CCV inhibitors as determined by in vitro immunocomplex kinase assay. Although CME is classically considered as a means to terminate signaling, our results demonstrate that SP-A uptake via CME by AMφ has to precede the initiation of SP-A signaling.
|Journal||American Journal of Physiology - Lung Cellular and Molecular Physiology|
|Number of pages||12|
|Publication status||Published - 03.2009|