sepharose and Emphysema

Pulmonary surfactant protein-D (SP-D) is a member of the collectin family of C-type lectins that is synthesized in many tissues including respiratory epithelial cells in the lung. SP-D is assembled predominantly as dodecamers consisting of four homotrimeric subunits each. Association of these subunits is stabilized by interchain disulfide bonds involving two conserved amino-terminal cysteine residues (Cys-15 and Cys-20). Mutant recombinant rat SP-D lacking these residues (RrSP-Dser15/20) is secreted in cell culture as trimeric subunits rather than as dodecamers. In this study, transgenic mice that express this mutant were generated to elucidate the functional importance of SP-D oligomerization in vivo. Expression of RrSP-Dser15/20 failed to correct the pulmonary phospholipid accumulation and emphysema characteristic of SP-D null (mSP-D-/-) mice. Expression of high concentrations of the mutant protein in wild-type mice reduced the abundance of disulfide cross-linked oligomers of endogenous SP-D in the bronchoalveolar lavage fluid and demonstrated a phenotype that partially overlapped with that of the SP-D-/- mice; the animals developed emphysema and foamy macrophages without the associated abnormalities in alveolar phospholipids typical of SP-D-/- mice. Development of foamy macrophages in SP-D-deficient mice is not secondary to the increased abundance of surfactant phospholipids. Disulfide cross-linked SP-D oligomers are required for the regulation of surfactant phospholipid homeostasis and the prevention of emphysema and foamy macrophages in vivo.

Topics: Animals; Base Sequence; Blotting, Western; Bronchoalveolar Lavage Fluid; Cysteine; Dimerization; Disulfides; DNA, Complementary; Dose-Response Relationship, Drug; Emphysema; Genotype; Glycoproteins; Immunoblotting; Lectins; Lung; Macrophages; Mice; Mice, Transgenic; Molecular Sequence Data; Mutation; Phenotype; Phosphatidylcholines; Protein Binding; Protein Conformation; Pulmonary Surfactant-Associated Protein D; Pulmonary Surfactants; Rats; Recombinant Proteins; Sepharose