Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Production of H5N1 avian influenza triple H5N1/NA-HA-M1 VLPs. ( A ) Schematic representation of the tricistronic expression cassette. ( B ) Western blotting analysis of NA-HA-M1 VLPs purified on a 10%–60% sucrose gradient. Antibodies used for the detection of HA, NA and M1 proteins: mouse monoclonal anti-HA H5N1 antibodies, mouse monoclonal anti-M1 influenza antibodies and rabbit polyclonal anti-avian influenza A neuraminidase antibody. ( C ) Schematic representation of the predicted structure of triple H5N1/NA-HA-M1 VLPs. Structural proteins are color-coded according to the scheme of the expression cassette: NA—yellow, HA—blue, M1—green.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Expressing, Western Blot, Purification

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Characterization of H5N1 avian influenza virus triple H5N1/NA-HA-M1 VLPs. ( A ) Transmission electron microscopy of triple H5N1/NA-HA-M1 VLPs and influenza A/H5N2 virus. Scale bar = 1 µm is shown in the right bottom corner of the images. ( B ) Hemagglutination assay. The AIV A/ostrich/Denmark/725/96 (H5N2) was used as a positive control. The HA titer was determined as the reciprocal of the highest dilution with HA activity. Each dot represents HA titers obtained in each experiment. The bars represent the median values of obtained HA titers. ( C ) Neuraminidase activity assay. The AIV A/Ck/Scot/59 (H5N1) was used as a positive control. For each assay, the mean value from three independent experiments performed is presented. The mean A560 values and standard deviations are shown on the y-axis.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Virus, Transmission Assay, Electron Microscopy, Hemagglutination Assay, Positive Control, Activity Assay

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Schematic timeline of immunization of the broiler hens with triple H5N1/NA-HA-M1 VLPs. Red droplets represent the days of blood collection.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques:

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: End-point titration of chicken sera after immunization with triple H5N1/NA-HA-M1 VLPs. ELISA plates were coated with reference antigen inactivated H5N1 virus. Serial dilutions of chicken A/H5N1/HPAI polyclonal antibodies served as a positive control. Chicken serum from day 0 and control chicken serum served as a background. The dilution factor of the pooled sera is shown on the x-axis. For each ELISA, the mean value from three independent experiments performed is presented. The mean A450 values and standard deviations are shown on the y-axis.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Titration, Enzyme-linked Immunosorbent Assay, Virus, Positive Control, Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Dynamics of anti-H5N1 IgY level in sera of immunized chickens. Kinetics of antibody titer in chickens ( n = 5) following prime and boost vaccinations with triple H5N1/NA-HA-M1 VLPs were measured via ELISA test. The median (thick line) is shown with the interquartile 25% and 75% range (narrow lines). The day of the serum collection is shown on the x-axis. The A450 values are shown on the y-axis. Statistical analysis was performed using the nonparametric Kruskal–Wallis test ( p = 0.05) and Benjamini, Krieger and Yekutieli multiple comparison test ( p = 0.05). Statistical differences were detected between days 0–41 and 0–34 ( p = 0.014) and shown on the graph as a star symbol.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: HI titer of pooled chicken sera collected after immunization with triple H5N1/NA-HA-M1 VLPs. H5N1 A/Ck/Scot/59 and H5N2 A/Ost/Den/72420/96 antibodies were used as a positive control. Sera from chickens vaccinated with PBS/ICF mixture served as a negative control. HI assay was performed in triplicates.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Positive Control, Negative Control, HI Assay

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Construction and characterization of HA-stalk universal influenza antigen from homologous H5N1 HPAI virus strain. ( A ) Schematic representation of the full-length HA (top panel) and HA-stalk constructs (bottom panel). To obtain the HA-stalk construct, a glycine linker was added between cysteines in positions C52 and C277, replacing the head region of the HA1 domain. ( B ) Schematic representation of the predicted structure of HA monomer and HA-stalk monomer. ( C ) Expression of H5N1 HA-stalk in insect cells was confirmed by IPMA with anti-H5N1 monoclonal antibodies. Full-length HA from the H5N1 strain was used as a positive control. Cells infected with the wild type baculovirus were used as a negative control. Images were taken at ×10 magnification. ( D ) Reactivity of the HA-stalk antigen and full-length HA from H5N1 in the IPMA with broadly neutralizing universal FI6 human antibodies. HA-stalk and full-length HA from the H5N1 strain was detected in transfected insect cells. Images were taken at ×20 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Virus, Construct, Expressing, Positive Control, Infection, Negative Control, Transfection

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Expression of HA-stalk antigens from the 1 and 2 HA groups in mammalian cells. HEK293 cells were transfected with HA-stalk H1N1, H5N1 and H7N9 constructs. Protein expression was detected using different monoclonal antibodies specific for H1N1, H5N1 and H7N9 influenza strains. Non-transfected cells were used as a negative control. Images were taken at ×40 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Expressing, Transfection, Construct, Negative Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Cross-reactivity of chicken sera obtained after vaccination with triple H5N1/NA-HA-M1 VLPs with the HA-stalk antigens from the 1 and 2 HA groups. Detection of HA-stalk from H1N1, H5N1 and H7N9 strains was performed on transfected HEK293 cells. Sera from unvaccinated hens were used as a background. Non-transfected cells were used as a negative control. Images were taken at ×40 magnification.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Transfection, Negative Control

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Alignment of amino acid sequences coding LAH regions from H5N1, pH1N1, H7N9 and H3N2 HA protein. ( A ) Alignment view with consensus sequence where the highest similarity is shown as a green colour. ( B ) Matrix showing the percentage of sequence identity between sequences.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Sequencing

Journal: Viruses

Article Title: Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

doi: 10.3390/v14040730

Figure Lengend Snippet: Cross-reactivity of chicken sera obtained after vaccination with triple H5N1/NA-HA-M1 VLPs with the LAH peptide from H3 from 2 HA group. The antibody titer in chickens ( n = 5) before (gray) and after (black) vaccination with triple H5N1/NA-HA-M1 VLPs was measured via the peptide ELISA test. The mean OD values and standard deviations are shown on the y-axis. The tested chickens are shown on the x-axis. Statistical analysis was performed using a nonparametric Wilcoxon test ( p = 0.05) for paired groups.

Article Snippet: Antibodies: Anti-H5N1 virus A/Ck/Scot/59 polyclonal chicken antibodies (cat. No RAA7002, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-H5N2 virus A/Ost/Den/72420/96 polyclonal chicken antibodies (cat. No RAA7003, Animal Health and Veterinary Laboratories Agency, Weybridge, UK); anti-M1 influenza mouse monoclonal (cat. No ab22396, Abcam Inc., Waltham, MA USA); anti-avian influenza A neuraminidase antibody (cat. No ab21304, Abcam Inc., Waltham, MA USA); anti-H1N1 mouse monoclonal antibodies (cat. No 11048-MM08, SinoBiological Inc., Beijing, China); FI6—highly specific humanized synthetic universal neutralizing monoclonal antibodies selected from plasma cells that bind group 1 and 2 influenza A HA (kind gift from Dr. Krzysztof Lacek and Alfredo Nicosia); anti-HA H5N1 mouse monoclonal antibodies (cat. No 11048-MM06, SinoBiological Inc., Beijing, China); anti-HA H3N2 mouse monoclonal antibodies (cat. No 11056-MM03, SinoBiological Inc., Beijing, China); anti-HA H7N9 mouse monoclonal antibodies (cat. No 11082-MM04, SinoBiological Inc., Beijing, China).

Techniques: Peptide ELISA

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Schematic overview of the anti-SARS-CoV-2 drug screening procedure. ( a ) ACE2 targeted inhibitors and compounds from the natural product library were applied prior to infection with SARS-CoV-2 (pre-infection treatment) while compounds from the FDA-approved library and the flavonoid library were applied post infection (post- infection treatment). Out of 2191 compounds tested, 121 displayed viral CPE reductions and 7 were selected for further validations using Vero E6 and HuH7 cell lines and hNEC. ( b ) Uninfected cells, infected cells treated with 0.1% DMSO, and infected cells treated with remdesivir were included as controls in the screen. Following treatment with citicoline, pravastatin sodium, tenofovir alafenamide, imatinib mesylate, calcitriol, dexlansoprazole, and prochlorperazine dimaleate, reduced CPE was observed when compared to the DMSO control.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Drug discovery, Infection, Control

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Summary of compounds identified from the primary screen that exhibit potential antiviral effects against SARS-CoV-2 infection.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Infection, Reverse Transcription, Reflux

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Validation of primary hits. For validating compounds with activity pre-infection, Vero E6 cells were first pre-treated with increasing concentrations of ( a ) citicoline, ( b ) pravastatin sodium and ( c ) tenofovir alafenamide prior to infection with SARS-CoV-2. Similarly, Huh7 cells were pre-treated with ( d ) citicoline, ( e ) pravastatin sodium and ( f ) tenofovir alafenamide and subsequently infected with SARS-CoV-2. For post-infection treatment validation, Vero E6 cells were infected with SARS-CoV-2 and treated with increasing concentrations of ( g ) imatinib mesylate, ( h ) calcitriol, ( i ) dexlansoprazole and ( j ) prochlorperazine dimaleate. Similarly, HuH7 cells were also infected then treated with a range of concentrations of ( k ) imatinib mesylate, ( l ) calcitriol, ( m ) dexlansoprazole and ( n ) prochlorperazine dimaleate. The primary and secondary exes correspond to the viral titre and relative cell viability, and the dashed line represents the CC 50 cut-off for cell viability. One-way ANOVA and Dunnett’s post-test were used to determine statistical differences, with * denoting p < 0.05, ** denoting p < 0.01 and *** denoting p < 0.001. Error bars represent the standard deviation observed from the means of triplicates performed for both cell viability and dose-dependent inhibition studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Biomarker Discovery, Activity Assay, Infection, Standard Deviation, Inhibition

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Validation of primary hits in hNECs. Selected primary hits were further validated in hNECs. Cells were either pre-treated with 10 μM citicoline prior to SARS-CoV-2 infection or treated following SARS-CoV-2 infection with 10 μM imatinib mesylate and 10 μM calcitriol. One-way ANOVA and Dunnett’s post-test were used to determine statistical differences, with ** denoting that p < 0.01. Error bars represent the standard deviation observed from the means of triplicates performed for dose-dependent inhibition studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Biomarker Discovery, Infection, Standard Deviation, Inhibition

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: SARS-CoV-2 and calcitriol modulate vitamin D Receptor (VDR), 24-hydroxylase (24(OH)ase), and cathelicidin (LL-37) in Vero E6 and Huh7 cells. In the first part of the experiment, Vero E6 and Huh7 cells were either mock-infected with media, or SARS-CoV-2 infected and harvested at indicated timepoints in the absence of calcitriol for baseline VDR, 24(OH)ase, and LL-37 mRNA expression. Vero E6 expression of ( a ) VDR, ( b ) 24(OH)ase and ( c ) LL-37 and Huh7 expression of ( d ) VDR, ( e ) 24(OH)ase and ( f ) LL-37 were as shown. In the second part of the experiment, Vero E6 and Huh7 cells were either mock-infected with media or SARS-CoV-2 for 1 h as above, then treated with 0.01, 0.1, 1, 10 or 20 µM calcitriol following infection. At 48 h post infection, the cells were harvested to check for VDR, 24(OH)ase, and LL-37 mRNA expression. Vero E6 mRNA expression of ( g ) VDR, ( i ) 24(OH)ase and ( k ) LL-37 and Huh7 expression of ( h ) VDR, ( j ) 24(OH)ase and ( l ) LL-37 were as shown. Data are represented as relative quantity to 0.1% DMSO control, or as relative quantity to mock-infected control. One-way ANOVA and Dunnett’s post-test and paired t -test were used to determine statistical differences, with * denoting p < 0.05, ** denoting p < 0.01 and *** denoting p < 0.001. Error bars represent the standard deviation observed from the means of triplicates performed for mRNA expression studies.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Infection, Expressing, Control, Standard Deviation

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: Western blot analysis of SARS-CoV-2 spike and VDR proteins in Vero E6 cells. ( a ) Vero E6 cells are either mock infected with media (−) or with SARS-CoV-2 (+) for 1 h. After 1 h post-infection, the cells are then either untreated (−) or treated (+) with 20 µM calcitriol for 6-, 24- and 48- hours before harvesting of total cell lysate. ( b ) Densitometry quantitation of protein expression levels is shown as fold changes to beta-actin. Full-length blots are shown in .

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: Western Blot, Infection, Quantitation Assay, Expressing

Journal: Pharmaceutics

Article Title: Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants

doi: 10.3390/pharmaceutics15030925

Figure Lengend Snippet: In vivo study of calcitriol treatment. 8-week old K18-hACE2 male and female mice were separated into treatment and non-treatment groups. Treatment study group has been given 3 days pre-treatment and 5 days post-treatment of calcitriol (5 μg/kg) via intraperitoneal injection. The non-treatment group has been given a PBS as a treatment control. Next, 10 3 PFU of SARS-CoV-2 (L, Alpha, and Beta variants) were inoculated through intranasal delivery to all groups after pre-treatment. Physiological parameters: body weight ( a – c ), survival rate ( d – f ), and physiological conditions were monitored after infections. For comparison between individual physiological conditions, 5 dpi infected mice were compared between treatment groups of all three variant infected mice ( g – i ). Scoring of physiological conditions was based on five criteria: appearance of the mouse coat, level of consciousness, activity level, eye condition, and respiratory quality. To compare the severity of damage in mouse tissues after infection, left lung lobes from 4 dpi mice were processed for histological analyses and scored based on six criteria to determine severity ( j – l ). The six criteria are inflammatory cell infiltration, haemorrhage, oedema, bronchial epithelial cell damage, degeneration of alveolar epithelial cells, and parenchymal wall expansion. For viral load determination, right lung, brain, liver, and spleen tissues from the same 4 dpi mice were harvested and homogenised for virus titration ( m – o ). Data is not shown for liver and spleen tissues. Statistical significance is determined with a two-tailed unpaired t -test, with * denoting p < 0.05. Survival group: variant L: n = 6 (treatment and no treatment); Alpha: n = 6 (treatment), n = 4 (no treatment); Beta: n = 6 (treatment), n = 5 (no treatment). 4 dpi group: variant L: n = 7 (treatment and no treatment); Alpha and Beta: n = 5 each (treatment and no treatment). Mock infection, n = 3.

Article Snippet: A primary screen to identify novel compounds with potential antiviral effects against SARS-CoV-2 infection was performed on four different libraries, namely a 462-compound ACE2-targeted compound library (CADD) (TargetMol, Boston, MA, USA), a 57-compound natural product library, a 500-compound flavonoids library (TimTec, Tampa, FL, USA), and a 1172-compound FDA-approved drug library (Selleckchem, Houston, TX, USA).

Techniques: In Vivo, Injection, Control, Comparison, Infection, Variant Assay, Activity Assay, Virus, Titration, Two Tailed Test