Protective role of the lung collectins surfactant protein A and surfactant protein D in airway inflammation  Angela Haczku, MD, PhD  Journal of Allergy.

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Protective role of the lung collectins surfactant protein A and surfactant protein D in airway inflammation  Angela Haczku, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 122, Issue 5, Pages 861-879 (November 2008) DOI: 10.1016/j.jaci.2008.10.014 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Lung collectins are produced by epithelial cells of the distal and proximal air spaces. A, Photomicrograph of a type II alveolar epithelial cell. Although the lung collectins have been localized to type II alveolar epithelial cells, immunocytochemistry studies indicate that they do not colocalize with lamellar bodies, the subcellular organelles responsible for storage and release of surfactant. Lamellar bodies were labeled with a monoclonal anti–ATP-binding cassette, subfamily A, member 3 antibody, 3C9 (Texas red, left panel). SP-D was labeled by a rabbit polyclonal antimouse SP-D (fluorescein isothiocyanate, second from left). The merged magnified image (right panel) illustrates that SP-D is scattered in the cytoplasm in a diffuse, reticular pattern and is localized outside the lamellar bodies. B, SP-A and SP-D are released onto mucosal surfaces to fulfill innate immune regulatory functions. Alveolar type II epithelial cells and Clara cells constitutively release SP-A and SP-D onto the luminal surface, where lung collectins associate with alveolar macrophages. C, SP-D expression is prominent in both the alveoli and proximal airway epithelial cells. Collectins can be detected not only in the distal but also in the proximal airways, and their expression can be shifted during the inflammatory airway response to the site of inflammation involving Clara cells and goblet cells. The photomicrograph shows epithelial cells of a proximal airway and type II alveolar epithelial cells both positively labeled with a rabbit polyclonal anti–SP-D (fluorescein isothiocyanate). Journal of Allergy and Clinical Immunology 2008 122, 861-879DOI: (10.1016/j.jaci.2008.10.014) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 The SP-A and SP-D genes are under differential inflammatory regulation. A, Allergic sensitization and challenge with A fumigatus induced increased SP-D levels in the BAL fluid. Levels of SP-A did not change. SP-B and SP-C levels decreased by ∼50%. Western blot analysis of the small (SP-A, SP-D) and large (SP-B, SP-C) aggregate surfactant fractions. B, Murine recombinant IL-4 (1.5 μg) induced a time-dependent increase in SP-D mRNA and protein expression in sensitized BALB/c mice in vivo. C, IL-13 (but not IFN-γ [not shown]) induced intracellular SP-D protein increase in isolated type II alveolar epithelial cells in vitro. D, Schematic drawing of the murine SP-D promoter illustrates proximal location of C/EBP, NFAT, AP-1, and STAT binding sites. The proximal SP-D promoter shows an almost perfect alignment between mice and rats and with the exception of the STAT binding site, a close homology with the human promoter. Journal of Allergy and Clinical Immunology 2008 122, 861-879DOI: (10.1016/j.jaci.2008.10.014) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 The collectin structure. A, The collectin triple helix. The collectins are assembled as trimeric subunits of a CRD, neck, and collagen region. B, SP-A, MBL and C1q have relatively short collagen domains, with a configuration resembling a bouquet of tulips. C, SP-D and conglutinin have a characteristic cruciform structure, assembled as tetramers of 3 identical polypeptides. Journal of Allergy and Clinical Immunology 2008 122, 861-879DOI: (10.1016/j.jaci.2008.10.014) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 The “head or tail hypothesis”185: collectins are capable of differential binding through either their CRD or their collagen domain to cell membrane receptors and eliciting respective anti-inflammatory or proinflammatory signaling pathways. A, In the absence of pathogens, collectins bind to the innate immune cells with their head and deliver inhibitory signals. B, Collectin binding to pathogens via CRD and presenting the collagen tail to phagocytes induce proinflammatory cytokine release. DC, Dendritic cell. Journal of Allergy and Clinical Immunology 2008 122, 861-879DOI: (10.1016/j.jaci.2008.10.014) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Lung collectins play an immunoprotective role during resolution of the inflammatory airway response at multiple levels. A, Collectins are capable of inhibiting dendritic cell maturation and activation. B, Collectins also inhibit T cells and the consequent activation of the adaptive immune response, immunoglobulin production by B cells and influx of granulocytes. Collectins inhibit mediator release directly from inflammatory cells. C, Collectins facilitate removal of apoptotic cells during resolution of the inflammatory response. Journal of Allergy and Clinical Immunology 2008 122, 861-879DOI: (10.1016/j.jaci.2008.10.014) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions