Complementation of pulmonary abnormalities in SP-D(-/-) mice with an SP-D/conglutinin fusion protein.
Surfactant protein D (SP-D) and serum conglutinin are closely related members of the collectin family of host defense lectins. Although normally synthesized at different anatomic sites, both proteins participate in the innate immune response to microbial challenge. To discern the roles of specific domains in the function of SP-D in vivo, a fusion protein (SP-D/Cong(neck+CRD)) consisting of the NH(2)-terminal and collagenous domains of rat SP-D (rSP-D) and the neck and carbohydrate recognition domains (CRDs) of bovine conglutinin (Cong) was expressed in the respiratory epithelium of SP-D gene-targeted (SP-D(-/-)) mice. While SP-D/Cong(neck+CRD) fusion protein did not affect lung morphology and surfactant phospholipid levels in the lungs of wild type mice, the chimeric protein substantially corrected the increased lung phospholipids in SP-D(-/-) mice. The SP-D/Cong(neck+CRD) fusion protein also completely corrected defects in influenza A clearance and inhibited the exaggerated inflammatory response that occurs following viral infection. However, the chimeric protein did not ameliorate the ongoing lung inflammation, enhanced metalloproteinase expression, and alveolar destruction that characterize this model of SP-D deficiency. By contrast, a single arm mutant (RrSP-D(Ser15,20)) partially restored antiviral activity but otherwise failed to rescue the deficient phenotype. Our findings directly implicate the CRDs of both SP-D and conglutinin in host defense in vivo. Our findings also strongly suggest that the molecular mechanisms underlying impaired pulmonary host defense and abnormal lipid metabolism are distinct from those that promote ongoing inflammation and the development of emphysema.
Zhang
Liqian
L
Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039, USA.
Hartshorn
Kevan L
KL
Crouch
Erika C
EC
Ikegami
Machiko
M
Whitsett
Jeffrey A
JA
D000280:Administration, Inhalation; D000818:Animals; D015153:Blotting, Western; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D050260:Carbohydrate Metabolism; D002417:Cattle; D037222:Collectins; D016207:Cytokines / Q000096:biosynthesis; D018076:DNA, Complementary / Q000378:metabolism; D004646:Emphysema / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics* / Q000502:physiology*; D009980:Influenza A virus / Q000235:genetics; D008168:Lung / Q000002:abnormalities*; D016676:Macrophages, Alveolar / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D010713:Phosphatidylcholines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D037663:Pulmonary Surfactant-Associated Protein D; D011663:Pulmonary Surfactants / Q000235:genetics* / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012712:Serum Globulins / Q000235:genetics* / Q000502:physiology*
D037222:Collectins; D016207:Cytokines; D018076:DNA, Complementary; D006023:Glycoproteins; D010713:Phosphatidylcholines; D010743:Phospholipids; D037663:Pulmonary Surfactant-Associated Protein D; D011663:Pulmonary Surfactants; D011993:Recombinant Fusion Proteins; D012712:Serum Globulins; C024014:conglutinin