Hypothiocyanite and Hypothiocyanite/Lactoferrin Mixture Exhibit Virucidal Activity In Vitro against SARS-CoV-2
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Abstract
SARS-CoV-2 replicates efficiently in the upper airways during the prodromal stage, resulting in environmental viral shedding from patients with active COVID-19 as well as from asymptomatic individuals. There is a need to find pharmacological interventions to mitigate the spread of COVID-19. Hypothiocyanite and lactoferrin are molecules of the innate immune system with a large spectrum cidal activity. The Food and Drug Administration and the European Medicines Agency designated the hypothiocyanite and lactoferrin combination as an orphan drug. We report an in vitro study showing that micromolar concentrations of hypothiocyanite exhibit dose- and time-dependent virucidal activity against SARS-CoV-2 and that the latter is slightly enhanced by the simultaneous presence of lactoferrin.
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SciScore for 10.1101/2020.11.17.387571: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication Contamination: Cells were maintained in a 5% CO2 incubator at 37□°C, routinely checked for mycoplasma and confirmed negative. Table 2: Resources
Antibodies Sentences Resources Immune staining was performed by incubation with 1:3000 anti-VSV-M [23H12] monoclonal antibody (Kerafast) on the infected cells for 90 minutes at 37°C, followed by incubation with 1:1000 anti-rabbit Alexa Fluor® 488 (Thermo Fisher Scientific) for 60 minutes at 37°C. anti-VSV-M [ 23H12suggested: Noneanti-rabbitsuggested: None… SciScore for 10.1101/2020.11.17.387571: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication Contamination: Cells were maintained in a 5% CO2 incubator at 37□°C, routinely checked for mycoplasma and confirmed negative. Table 2: Resources
Antibodies Sentences Resources Immune staining was performed by incubation with 1:3000 anti-VSV-M [23H12] monoclonal antibody (Kerafast) on the infected cells for 90 minutes at 37°C, followed by incubation with 1:1000 anti-rabbit Alexa Fluor® 488 (Thermo Fisher Scientific) for 60 minutes at 37°C. anti-VSV-M [ 23H12suggested: Noneanti-rabbitsuggested: NoneExperimental Models: Cell Lines Sentences Resources Cell culture and viruses: Vero (African green monkey kidney cells, ATCC® CCL-81), Vero E6 (ATCC® CRL1586) and HEK293T (ATCC® CRL-11268TM) cells were grown in Dulbecco’s modified Eagle’s medium (D-MEM) containing 10% heat-inactivated fetal bovine serum (FBSi). Vero E6suggested: NoneTenfold virus dilutions were prepared in DMEM and inoculated on confluent Vero-E6 cells for 1 h at 37 °C. Vero-E6suggested: NonerVSV-S), HEK293T cells were seeded in T175 flasks and then transfected by calcium phosphate-DNA preciptation with 40 μg of pSARS-CoV-2-spike plasmid. HEK293Tsuggested: NoneViral stock was tenfold serially diluted in DMEM and inoculated on confluent Vero cells for 1 h at 37 °C. Verosuggested: NoneSoftware and Algorithms Sentences Resources Statistical analysis was carried out with GraphPad Prism the one-way ANOVA test. GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Curve fitting was performed to determine IC50 values using a sigmoidal 4PL model in GraphPad Prism 8 software. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- No funding statement was detected.
- No protocol registration statement was detected.
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