Rising, Anna
- Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
- Karolinska institutet
Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis and hospitalisation of infants in developed countries. Surfactant protein A (SP-A) is an important innate immune molecule, localized in pulmonary surfactant. SP-A binds to carbohydrates on the surface of pathogens in a calcium-dependent manner to enable neutralisation, agglutination and clearance of pathogens including RSV.SP-A forms trimeric units and further oligomerises through interactions between its N-terminal domains. Whilst a recombinant trimeric fragment of the closely related molecule (surfactant protein D) has been shown to retain many of the native protein's functions, the importance of the SP-A oligomeric structure in its interaction with RSV has not been determined.The aim of this study was to produce a functional trimeric recombinant fragment of human (rfh)SP-A, which lacks the N-terminal domain (and the capacity to oligomerise) and test its ability to neutralise RSV in an in vitro model of human bronchial epithelial infection.We used a novel expression tag derived from spider silk proteins ('NT') to produce rfhSP-A in Escherichia coli, which we found to be trimeric and to bind to mannan in a calcium-dependent manner. Trimeric rfhSP-A reduced infection levels of human bronchial epithelial (AALEB) cells by RSV by up to a mean (+/- SD) of 96.4 (+/- 1.9) % at 5 mu g/ml, which was significantly more effective than dimeric rfhSP-A (34.3 (+/- 20.5) %) (p
Surfactant protein A; Recombinant trimeric fragment; Respiratory syncytial virus; NT domain; Innate immunity; Neutralisation; Collectin
Immunobiology
2017, volym: 222, nummer: 2, sidor: 111-118
Utgivare: ELSEVIER GMBH, URBAN & FISCHER VERLAG
Biokemi
Molekylärbiologi
https://res.slu.se/id/publ/80755