Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2
SARS-CoV-2 infection of host cells is driven by binding of the SARS-CoV-2 spike-(S)-protein to lung type II pneumocytes, followed by virus replication. Surfactant protein SP-D, member of the front-line immune defense of the lungs, binds glycosylated structures on invading pathogens such as viruses t...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2021-07-01
|
Series: | Biomolecules |
Subjects: | |
Online Access: | https://www.mdpi.com/2218-273X/11/8/1114 |
id |
doaj-1ce6ee2a5d014a70b42089eb87d344e5 |
---|---|
record_format |
Article |
spelling |
doaj-1ce6ee2a5d014a70b42089eb87d344e52021-08-26T13:33:35ZengMDPI AGBiomolecules2218-273X2021-07-01111114111410.3390/biom11081114Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2Raquel Arroyo0Shawn N. Grant1Miriam Colombo2Lucia Salvioni3Fabio Corsi4Marta Truffi5Davide Ottolina6Brett Hurst7Marc Salzberg8Davide Prosperi9Paul S. Kingma10Division of Neonatology and Pulmonary Biology, Cincinnati Children’s Hospital, Cincinnati, OH 45229, USAAirway Therapeutics Inc., Cincinnati, OH 45249, USANanoBio laboratory, Department of Biotechnology and Bioscience, University of Milano-Bicocca, 20126 Milano, ItalyNanoBio laboratory, Department of Biotechnology and Bioscience, University of Milano-Bicocca, 20126 Milano, ItalyIstituti Clinici Scientifici Maugeri IRCCS, via Maugeri 4, 27100 Pavia, ItalyIstituti Clinici Scientifici Maugeri IRCCS, via Maugeri 4, 27100 Pavia, ItalyDivision of Anesthesiology and Intensive Care Medicine, ASST Fatebenefratelli Sacco, Luigi Sacco Hospital, University of Milan, 20157 Milano, ItalyInstitute for Antiviral Research, Utah State University, Logan, UT 84322, USAAirway Therapeutics Inc., Cincinnati, OH 45249, USANanoBio laboratory, Department of Biotechnology and Bioscience, University of Milano-Bicocca, 20126 Milano, ItalyDivision of Neonatology and Pulmonary Biology, Cincinnati Children’s Hospital, Cincinnati, OH 45229, USASARS-CoV-2 infection of host cells is driven by binding of the SARS-CoV-2 spike-(S)-protein to lung type II pneumocytes, followed by virus replication. Surfactant protein SP-D, member of the front-line immune defense of the lungs, binds glycosylated structures on invading pathogens such as viruses to induce their clearance from the lungs. The objective of this study is to measure the pulmonary SP-D levels in COVID-19 patients and demonstrate the activity of SP-D against SARS-CoV-2, opening the possibility of using SP-D as potential therapy for COVID-19 patients. Pulmonary SP-D concentrations were measured in bronchoalveolar lavage samples from patients with corona virus disease 2019 (COVID-19) by anti-SP-D ELISA. Binding assays were performed by ELISAs. Protein bridge and aggregation assays were performed by gel electrophoresis followed by silver staining and band densitometry. Viral replication was evaluated in vitro using epithelial Caco-2 cells. Results indicate that COVID-19 patients (<i>n</i> = 12) show decreased pulmonary levels of SP-D (median = 68.9 ng/mL) when compared to levels reported for healthy controls in literature. Binding assays demonstrate that SP-D binds the SARS-CoV-2 glycosylated spike-(S)-protein of different emerging clinical variants. Binding induces the formation of protein bridges, the critical step of viral aggregation to facilitate its clearance. SP-D inhibits SARS-CoV-2 replication in Caco-2 cells (EC<sub>90</sub> = 3.7 μg/mL). Therefore, SP-D recognizes and binds to the spike-(S)-protein of SARS-CoV-2 in vitro, initiates the aggregation, and inhibits viral replication in cells. Combined with the low levels of SP-D observed in COVID-19 patients, these results suggest that SP-D is important in the immune response to SARS-CoV-2 and that rhSP-D supplementation has the potential to be a novel class of anti-viral that will target SARS-CoV-2 infection.https://www.mdpi.com/2218-273X/11/8/1114SP-DCOVID-19spike-proteinACE2anti-inflammatory |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Raquel Arroyo Shawn N. Grant Miriam Colombo Lucia Salvioni Fabio Corsi Marta Truffi Davide Ottolina Brett Hurst Marc Salzberg Davide Prosperi Paul S. Kingma |
spellingShingle |
Raquel Arroyo Shawn N. Grant Miriam Colombo Lucia Salvioni Fabio Corsi Marta Truffi Davide Ottolina Brett Hurst Marc Salzberg Davide Prosperi Paul S. Kingma Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 Biomolecules SP-D COVID-19 spike-protein ACE2 anti-inflammatory |
author_facet |
Raquel Arroyo Shawn N. Grant Miriam Colombo Lucia Salvioni Fabio Corsi Marta Truffi Davide Ottolina Brett Hurst Marc Salzberg Davide Prosperi Paul S. Kingma |
author_sort |
Raquel Arroyo |
title |
Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 |
title_short |
Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 |
title_full |
Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 |
title_fullStr |
Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 |
title_full_unstemmed |
Full-Length Recombinant hSP-D Binds and Inhibits SARS-CoV-2 |
title_sort |
full-length recombinant hsp-d binds and inhibits sars-cov-2 |
publisher |
MDPI AG |
series |
Biomolecules |
issn |
2218-273X |
publishDate |
2021-07-01 |
description |
SARS-CoV-2 infection of host cells is driven by binding of the SARS-CoV-2 spike-(S)-protein to lung type II pneumocytes, followed by virus replication. Surfactant protein SP-D, member of the front-line immune defense of the lungs, binds glycosylated structures on invading pathogens such as viruses to induce their clearance from the lungs. The objective of this study is to measure the pulmonary SP-D levels in COVID-19 patients and demonstrate the activity of SP-D against SARS-CoV-2, opening the possibility of using SP-D as potential therapy for COVID-19 patients. Pulmonary SP-D concentrations were measured in bronchoalveolar lavage samples from patients with corona virus disease 2019 (COVID-19) by anti-SP-D ELISA. Binding assays were performed by ELISAs. Protein bridge and aggregation assays were performed by gel electrophoresis followed by silver staining and band densitometry. Viral replication was evaluated in vitro using epithelial Caco-2 cells. Results indicate that COVID-19 patients (<i>n</i> = 12) show decreased pulmonary levels of SP-D (median = 68.9 ng/mL) when compared to levels reported for healthy controls in literature. Binding assays demonstrate that SP-D binds the SARS-CoV-2 glycosylated spike-(S)-protein of different emerging clinical variants. Binding induces the formation of protein bridges, the critical step of viral aggregation to facilitate its clearance. SP-D inhibits SARS-CoV-2 replication in Caco-2 cells (EC<sub>90</sub> = 3.7 μg/mL). Therefore, SP-D recognizes and binds to the spike-(S)-protein of SARS-CoV-2 in vitro, initiates the aggregation, and inhibits viral replication in cells. Combined with the low levels of SP-D observed in COVID-19 patients, these results suggest that SP-D is important in the immune response to SARS-CoV-2 and that rhSP-D supplementation has the potential to be a novel class of anti-viral that will target SARS-CoV-2 infection. |
topic |
SP-D COVID-19 spike-protein ACE2 anti-inflammatory |
url |
https://www.mdpi.com/2218-273X/11/8/1114 |
work_keys_str_mv |
AT raquelarroyo fulllengthrecombinanthspdbindsandinhibitssarscov2 AT shawnngrant fulllengthrecombinanthspdbindsandinhibitssarscov2 AT miriamcolombo fulllengthrecombinanthspdbindsandinhibitssarscov2 AT luciasalvioni fulllengthrecombinanthspdbindsandinhibitssarscov2 AT fabiocorsi fulllengthrecombinanthspdbindsandinhibitssarscov2 AT martatruffi fulllengthrecombinanthspdbindsandinhibitssarscov2 AT davideottolina fulllengthrecombinanthspdbindsandinhibitssarscov2 AT bretthurst fulllengthrecombinanthspdbindsandinhibitssarscov2 AT marcsalzberg fulllengthrecombinanthspdbindsandinhibitssarscov2 AT davideprosperi fulllengthrecombinanthspdbindsandinhibitssarscov2 AT paulskingma fulllengthrecombinanthspdbindsandinhibitssarscov2 |
_version_ |
1721194673171595264 |