Enhanced antiviral activity of human surfactant protein d by site-specific engineering of the carbohydrate recognition domain
Innate immunity is critical in the early containment of influenza A virus (IAV) infection and surfactant protein D (SP-D) plays a crucial role in innate defense against IAV in the lungs. Multivalent lectin-mediated interactions of SP-D with IAVs result in viral aggregation, reduced epithelial infection, and enhanced IAV clearance by phagocytic cells. Previous studies showed that porcine SP-D (pSP-D) exhibits distinct antiviral activity against IAV as compared to human SP-D (hSP-D), mainly due to key residues in the lectin domain of pSP-D that contribute to its profound neutralizing activity. These observations provided the basis for the design of a full-length recombinant mutant form of hSP-D, designated as “improved SP-D” (iSP-D). Inspired by pSP-D, the lectin domain of iSP-D has 5 amino acids replaced (Asp324Asn, Asp330Asn, Val251Glu, Lys287Gln, Glu289Lys) and 3 amino acids inserted (326Gly-Ser-Ser). Characterization of iSP-D revealed no major differences in protein assembly and saccharide binding selectivity as compared to hSP-D. However, hemagglutination inhibition measurements showed that iSP-D expressed strongly enhanced activity compared to hSP-D against 31 different IAV strains tested, including (pandemic) IAVs that were resistant for neutralization by hSP-D. Furthermore, iSP-D showed increased viral aggregation and enhanced protection of MDCK cells against infection by IAV. Importantly, prophylactic or therapeutic application of iSP-D decreased weight loss and reduced viral lung titers in a murine model of IAV infection using a clinical isolate of H1N1pdm09 virus. These studies demonstrate the potential of iSP-D as a novel human-based antiviral inhalation drug that may provide immediate protection against or recovery from respiratory (pandemic) IAV infections in humans.
|Keywords||Antiviral agent, Collectin, Influenza A virus, Innate immunity, Lung infection, Pandemics, Recombinant expression, Surfactant protein D|
|Persistent URL||dx.doi.org/10.3389/fimmu.2019.02476, hdl.handle.net/1765/121329|
|Journal||Frontiers in Immunology|
van Eijk, M, Hillaire, M.L.B, Rimmelzwaan, G.F, Rynkiewicz, M.J. (Michael J.), White, M.R. (Mitchell R.), Hartshorn, K.L. (Kevan L.), … Haagsman, H.P. (2019). Enhanced antiviral activity of human surfactant protein d by site-specific engineering of the carbohydrate recognition domain. Frontiers in Immunology, 10(OCT). doi:10.3389/fimmu.2019.02476