Abstract
Surfactant protein SP-D is a multimeric collagenous lectin, called collectin. SP-D is a multifunctional, pattern recognition innate immune molecule, which binds in a calcium dependent manner to an array of carbohydrates and lipids, thus offering resistance to invading pathogens, allergen challenge, and pulmonary inflammation. SP-D is predominantly found in the endoplasmic reticulum of type 2 pneumocytes and in the secretory granules of Clara or non-ciliated bronchiolar cells. The highest expression of SP-D is observed in the distal airways and alveoli. There is also an extra pulmonary existence of SP-D. The common sources of native full-length human SP-D are bronchoalveolar lavage (BAL) washings from normal or preferably patients suffering from alveolar proteinosis who overproduce SP-D in the lungs. Amniotic fluid collected at the term during parturition is another reasonable source. Here, we describe a simple and rapid method of purifying native SP-D away from SP-A which is also present in the same source. We also describe procedures of expressing and purifying a recombinant fragment of human SP-D (rhSP-D) comprising trimeric neck and carbohydrate recognition domains that has been shown to have therapeutic effects in murine models of allergy and infection.
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References
King RJ, Klass DJ, Gikas EG, Clements JA (1973) An uncommon phosphatidylcholine specific for surface-active material in canine lung. Am J Physiol 224(4):788–795
Glasser SW, Korfhagen TR, Weaver T, Pilot-Matias T, Fox JL, Whitsett JA (1987) cDNA and deduced amino acid sequence of human pulmonary surfactant-associated proteolipid SPL(Phe). Proc Natl Acad Sci U S A 84(12):4007–4011
Warr RG, Hawgood S, Buckley DI, Crisp TM, Schilling J, Benson BJ et al (1987) Low molecular weight human pulmonary surfactant protein (SP5): isolation, characterization, and cDNA and amino acid sequences. Proc Natl Acad Sci U S A 84(22):7915–7919
Weaver TE, Whitsett JA (1991) Function and regulation of expression of pulmonary surfactant-associated proteins. Biochem J 273(Pt 2):249–264
Kishore U, Reid KB (2001) Structures and functions of mammalian collectins. Results Probl Cell Differ 33:225–248
Weaver TE (1998) Synthesis, processing and secretion of surfactant proteins B and C. Biochim Biophys Acta 1408(2–3):173–179
Pérez-Gil J, Keough KM (1998) Interfacial properties of surfactant proteins. Biochim Biophys Acta 1408(2–3):203–217
Crouch E, Rust K, Veile R, Donis-Keller H, Grosso L (1993) Genomic organization of human surfactant protein D (SP-D). SP-D is encoded on chromosome 10q22.2-23.1. J Biol Chem 268(4):2976–2983
Lu J, Willis AC, Reid KB (1992) Purification, characterization and cDNA cloning of human lung surfactant protein D. Biochem J 284(Pt 3):795–802
Gardai SJ, Xiao YQ, Dickinson M, Nick JA, Voelker DR, Greene KE et al (2003) By binding SIRPalpha or calreticulin/CD91, lung collectins act as dual function surveillance molecules to suppress or enhance inflammation. Cell 115(1):13–23
Hartshorn KL, Crouch EC, White MR, Eggleton P, Tauber AI, Chang D et al (1994) Evidence for a protective role of pulmonary surfactant protein D (SP-D) against influenza A viruses. J Clin Invest 94(1):311–319
Kuan SF, Rust K, Crouch E (1992) Interactions of surfactant protein D with bacterial lipopolysaccharides. Surfactant protein D is an Escherichia coli-binding protein in bronchoalveolar lavage. J Clin Invest 90(1):97–106
Schelenz S, Malhotra R, Sim RB, Holmskov U, Bancroft GJ (1995) Binding of host collectins to the pathogenic yeast Cryptococcus neoformans: human surfactant protein D acts as an agglutinin for acapsular yeast cells. Infect Immun 63(9):3360–3366
Madan T, Kishore U, Shah A, Eggleton P, Strong P, Wang JY et al (1997) Lung surfactant proteins A and D can inhibit specific IgE binding to the allergens of Aspergillus fumigatus and block allergen-induced histamine release from human basophils. Clin Exp Immunol 110(2):241–249
Kishore U, Strong P, Perdikoulis MV, Reid KB (2001) A recombinant homotrimer, composed of the alpha helical neck region of human surfactant protein D and C1q B chain globular domain, is an inhibitor of the classical complement pathway. J Immunol 166(1):559–565
Wright JR (2005) Immunoregulatory functions of surfactant proteins. Nat Rev Immunol 5(1):58–68
Kishore U, Greenhough TJ, Waters P, Shrive AK, Ghai R, Kamran MF et al (2006) Surfactant proteins SP-A and SP-D: structure, function and receptors. Mol Immunol 43(9):1293–1315
Mahajan L, Madan T, Kamal N, Singh VK, Sim RB, Telang SD et al (2008) Recombinant surfactant protein-D selectively increases apoptosis in eosinophils of allergic asthmatics and enhances uptake of apoptotic eosinophils by macrophages. Int Immunol 20(8):993–1007
Herbein JF, Savov J, Wright JR (2000) Binding and uptake of surfactant protein D by freshly isolated rat alveolar type II cells. Am J Physiol Lung Cell Mol Physiol 278(4):L830–L839
Miyamura K, Leigh LE, Lu J, Hopkin J, López Bernal A, Reid KB (1994) Surfactant protein D binding to alveolar macrophages. Biochem J 300(Pt 1):237–242
Ogasawara Y, Kuroki Y, Akino T (1992) Pulmonary surfactant protein D specifically binds to phosphatidylinositol. J Biol Chem 267(29):21244–21249
Barnes P, Liew F (1995) Nitric oxide and asthmatic inflammation. Immunol Today 16(3): 128–130
Erpenbeck VJ, Schmidt R, Günther A, Krug N, Hohlfeld JM (2006) Surfactant protein levels in bronchoalveolar lavage after segmental allergen challenge in patients with asthma. Allergy 61(5):598–604
Wang JY, Kishore U, Lim BL, Strong P, Reid KB (1996) Interaction of human lung surfactant proteins A and D with mite (Dermatophagoides pteronyssinus) allergens. Clin Exp Immunol 106:367–373
Madan T, Reid KB, Singh M, Sarma PU, Kishore U (2005) Susceptibility of mice genetically deficient in the surfactant protein (SP)-A or SP-D gene to pulmonary hypersensitivity induced by antigens and allergens of Aspergillus fumigatus. J Immunol 174(11):6943–6954
Madan T, Kishore U, Singh M, Strong P, Hussain EM, Reid KB et al (2001) Protective role of lung surfactant protein D in a murine model of invasive pulmonary aspergillosis. Infect Immun 69(4):2728–2731
Singh M, Madan T, Waters P, Parida SK, Sarma PU, Kishore U (2003) Protective effects of a recombinant fragment of human surfactant protein D in a murine model of pulmonary hypersensitivity induced by dust mite allergens. Immunol Lett 86(3):299–307
Fisher JH, Mason R (1995) Expression of pulmonary surfactant protein D in rat gastric mucosa. Am J Respir Cell Mol Biol 12(1):13–18
Madsen J, Kliem A, Tornoe I, Skjodt K, Koch C, Holmskov U (2000) Localization of lung surfactant protein D on mucosal surfaces in human tissues. J Immunol 164(11):5866–5870
Herías MV, Hogenkamp A, van Asten AJ, Tersteeg MH, van Eijk M, Haagsman HP (2007) Expression sites of the collectin SP-D suggest its importance in first line host defence: power of combining in situ hybridisation. RT-PCR and immunohistochemistry. Mol Immunol 44(13):3324–3332
Lin Z, deMello D, Phelps D, Koltun W, Page M, Floros J (2001) Both human SP-A1 and Sp-A2 genes are expressed in small and large intestine. Pediatr Pathol Mol Med 5:367–386
Chailley-Heu B, Rubio S, Rougier J, Ducroc R, Barlier-Mur A, Ronco P et al (1997) Expression of hydrophilic surfactant proteins by mesentery cells in rat and man. Biochem J 328:251–256
Rubio S, Lacaze-Masmonteil T, Chailley-Heu B, Kahn A, Bourbon J, Ducroc R (1995) Pulmonary surfactant protein A (SP-A) is expressed by epithelial cells of small and large intestine. J Biol Chem 270:12162–12169
Madsen J, Tornoe I, Nielsen O, Koch C, Steinhilber W, Holmskov U (2003) Expression and localization of lung surfactant protein A in human tissues. Am J Respir Cell Mol Biol 29:591–597
Brauer L, Kindler C, Jäger K, Sel S, Nölle B, Pleyer U et al (2007) Detection of surfactant proteins A and D in human tear fluid and the human lacrimal system. Invest Ophthalmol Vis Sci 48:3945–3953
Honda Y, Kuroki Y, Matsuura E, Nagae H, Takahashi H, Akino T et al (1995) Pulmonary surfactant protein D in sera and bronchoalveolar lavage fluids. Am J Respir Crit Care Med 152(6 Pt 1):1860–1866
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Dodagatta-Marri, E. et al. (2014). Purification of Surfactant Protein D (SP-D) from Pooled Amniotic Fluid and Bronchoalveolar Lavage. In: Gadjeva, M. (eds) The Complement System. Methods in Molecular Biology, vol 1100. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-724-2_22
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DOI: https://doi.org/10.1007/978-1-62703-724-2_22
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