Interactions of recombinant human pulmonary surfactant protein D and SP-D multimers with influenza A.
To further study the structure and function of surfactant protein D (SP-D), recombinant human SP-D (rhSP-D) was isolated from the culture medium of Chinese hamster ovary (CHO)-K1 cells stably transfected with a full-length hSP-D cDNA. Although a significant fraction of the secreted rhSP-D was recovered as dodecamers similar to recombinant rat SP-D (rrSP-D), a major fraction accumulated as multimers of dodecamers indistinguishable from human proteinosis SP-D. As previously shown for the rat protein, rhSP-D agglutinated specific strains of influenza A virus (IAV), inhibited viral hemagglutinin activity, and protected neutrophils (PMN) from deactivation by IAV. However, the potency of rhSP-D multimers was severalfold greater than for purified dodecamers, comparable to natural, proteinosis hSP-D. Although rhSP-D multimers were also more potent than the serum collectins in mediating viral aggregation and protection of PMN, they were less potent than conglutinin in inhibiting infectivity in vitro. These studies establish that the propensity of hSP-D to form multimers of dodecamers is determined by its primary structure and demonstrate carbohydrate recognition domain valency-dependent interactions of SP-D with IAV.
Hartshorn
K
K
Department of Medicine, Boston University School of Medicine, Massachusetts 02118, USA.
Chang
D
D
Rust
K
K
White
M
M
Heuser
J
J
Crouch
E
E
D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins; D002645:Chickens; D002850:Chromatography, Gel; D006224:Cricetinae; D018076:DNA, Complementary; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D006384:Hemagglutination; D006801:Humans; D009980:Influenza A virus / Q000472:pathogenicity / Q000502:physiology*; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D010063:Ovum / Q000821:virology; D037663:Pulmonary Surfactant-Associated Protein D; D011663:Pulmonary Surfactants / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000648:ultrastructure; D014162:Transfection
D002352:Carrier Proteins; D018076:DNA, Complementary; D006023:Glycoproteins; D046911:Macromolecular Substances; D037663:Pulmonary Surfactant-Associated Protein D; D011663:Pulmonary Surfactants; D011994:Recombinant Proteins