A Recombinant Fragment of Human Surfactant Protein D Binds Spike Protein and Inhibits Infectivity and Replication of SARS-CoV-2 in Clinical Samples

Autor: Uday Kishore, Ekta Gupta, Hrishikesh Pandit, Indra Kundu, Sheetalnath Rooge, Savneet Kaur, Reshu Agarwal, Sanjeev Gupta, Taruna Madan, Dinesh M. Tripathi, Susan Idicula-Thomas, Praveen M. Varghese, Rambhadur Subedi, Barnali Biswas
Rok vydání: 2021
Předmět:
entry inhibition
Male
0301 basic medicine
viruses
Clinical Biochemistry
Plasma protein binding
Virus Replication
spike protein
medicine.disease_cause
law.invention
0302 clinical medicine
law
Chlorocebus aethiops
Receptor
skin and connective tissue diseases
Pulmonary Surfactant-Associated Protein D
Original Research
Coronavirus
Infectivity
Chemistry
virus diseases
Recombinant Proteins
Spike Glycoprotein
Coronavirus
Recombinant DNA
Female
Protein Binding
medicine.drug
Adult
Pulmonary and Respiratory Medicine
surfactant protein D
Protein subunit
Virus
03 medical and health sciences
medicine
Animals
Humans
Gene
Vero Cells
Molecular Biology
entry inhibitor
SARS-CoV-2
fungi
COVID-19
Surfactant protein D
Cell Biology
Molecular biology
Virology
Entry inhibitor
body regions
030104 developmental biology
030228 respiratory system
Viral replication
Zdroj: American Journal of Respiratory Cell and Molecular Biology
ISSN: 1535-4989
1044-1549
DOI: 10.1165/rcmb.2021-0005oc
Popis: RationaleCOVID-19 is an acute infectious disease caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Human surfactant protein D (SP-D) is known to interact with spike protein of SARS-CoV, but its immune-surveillance against SARS-CoV-2 is not known.ObjectiveThis study aimed to examine the potential of a recombinant fragment of human SP-D (rfhSP-D) as an inhibitor of replication and infection of SARS-CoV-2.MethodsrfhSP-D interaction with spike protein of SARS-CoV-2 and hACE-2 receptor was predicted via docking analysis. The inhibition of interaction between spike protein and ACE-2 by rfhSP-D was confirmed using direct and indirect ELISA. The effect of rfhSP-D on replication and infectivity of SARS-CoV-2 from clinical samples was studied by measuring the expression of RdRp gene of the virus using qPCR.Measurements and Main ResultsIn-silico interaction studies indicated that three amino acid residues in the RBD of spike of SARS-CoV-2 were commonly involved in interacting with rfhSP-D and ACE-2. Studies using clinical samples of SARS-CoV-2 positive cases (asymptomatic, n=7 and symptomatic, n=8 and negative controls n=15) demonstrated that treatment with 5μM rfhSP-D inhibited viral replication by ~5.5 fold and was more efficient than Remdesivir (100 μM). Approximately, a 2-fold reduction in viral infectivity was also observed after treatment with 5μM rfhSP-D.ConclusionsThese results conclusively demonstrate that the calcium independent rfhSP-D mediated inhibition of binding between the receptor binding domain of the S1 subunit of the SARS-CoV-2 spike protein and human ACE-2, its host cell receptor, and a significant reduction in SARS-CoV-2 infection and replication in-vitro.
Databáze: OpenAIRE
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