recombinant influenza a virus h5n1 hemagglutinin  (Sino Biological)


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    Name:
    Influenza A H5N1 Hemagglutinin HA1 Protein
    Description:
    A DNA sequence encoding the Influenza A virus A Hubei 1 2010 H5N1 hemagglutinin AEO89181 1 Met1 Arg339 termed as HA1 was expressed with a polyhistidine tag at the C terminus
    Catalog Number:
    40060-V08H1
    Price:
    None
    Category:
    recombinant protein
    Host:
    HEK293 Cells
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    Structured Review

    Sino Biological recombinant influenza a virus h5n1 hemagglutinin
    HA-based vaccines stimulate antigen-specific antibody responses in BALB/c mice. a Schematic representation of the standard immunization schedule (priming + two boosters). b Antigen-specific total IgG titer in sera collected on day 30 from BALB/c mice subjected to i.m. immunization with different antigens conjugated to 200 kDa HA or injected alone (standard immunization schedule; BSA, Vк 3-20 , OVA, n = 12; SOD, hGH, TT, RABV G, n = 6; <t>H5N1,</t> n = 4). c Anti-OVA total IgG and IgG subclass titers detected on day 30 in sera of BALB/c mice subjected to i.m. immunization with 10 μg of OVA alone, conjugated to HA, or emulsified with alum following the standard schedule ( n = 10 mice/group; HA vs. alum: P
    A DNA sequence encoding the Influenza A virus A Hubei 1 2010 H5N1 hemagglutinin AEO89181 1 Met1 Arg339 termed as HA1 was expressed with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/recombinant influenza a virus h5n1 hemagglutinin/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant influenza a virus h5n1 hemagglutinin - by Bioz Stars, 2021-09
    94/100 stars

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    1) Product Images from "Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines"

    Article Title: Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/s41423-021-00667-y

    HA-based vaccines stimulate antigen-specific antibody responses in BALB/c mice. a Schematic representation of the standard immunization schedule (priming + two boosters). b Antigen-specific total IgG titer in sera collected on day 30 from BALB/c mice subjected to i.m. immunization with different antigens conjugated to 200 kDa HA or injected alone (standard immunization schedule; BSA, Vк 3-20 , OVA, n = 12; SOD, hGH, TT, RABV G, n = 6; H5N1, n = 4). c Anti-OVA total IgG and IgG subclass titers detected on day 30 in sera of BALB/c mice subjected to i.m. immunization with 10 μg of OVA alone, conjugated to HA, or emulsified with alum following the standard schedule ( n = 10 mice/group; HA vs. alum: P
    Figure Legend Snippet: HA-based vaccines stimulate antigen-specific antibody responses in BALB/c mice. a Schematic representation of the standard immunization schedule (priming + two boosters). b Antigen-specific total IgG titer in sera collected on day 30 from BALB/c mice subjected to i.m. immunization with different antigens conjugated to 200 kDa HA or injected alone (standard immunization schedule; BSA, Vк 3-20 , OVA, n = 12; SOD, hGH, TT, RABV G, n = 6; H5N1, n = 4). c Anti-OVA total IgG and IgG subclass titers detected on day 30 in sera of BALB/c mice subjected to i.m. immunization with 10 μg of OVA alone, conjugated to HA, or emulsified with alum following the standard schedule ( n = 10 mice/group; HA vs. alum: P

    Techniques Used: Mouse Assay, Injection

    Related Articles

    Recombinant:

    Article Title: Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines
    Article Snippet: .. Different antigens, i.e. ovalbumin (OVA, Hyglos GmbH); human superoxide dismutase (SOD), bovine serum albumin (BSA), human growth hormone (hGH, Sigma-Aldrich); tetanus toxoid (TT, Alomone Labs); recombinant influenza A virus H5N1 hemagglutinin (H5N1, Sino Biological Inc.); a recombinant immunoglobulin (Ig) light κ chain variable region (Vκ3–20 , a kind gift from Prof. R. Dolcetti, Centro di Riferimento Oncologico di Aviano (CRO-IRCCS), Aviano, Italy); and rabies virus G glycoprotein (RABV G; obtained from Challenge Virus Strain—11 (ATCC® VR-959™), propagated in BHK-21 cells (ATCC® CCL-10™), and purified as previously described ), were conjugated to HA-acetal using a general procedure for each antigen. ..

    Purification:

    Article Title: Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines
    Article Snippet: .. Different antigens, i.e. ovalbumin (OVA, Hyglos GmbH); human superoxide dismutase (SOD), bovine serum albumin (BSA), human growth hormone (hGH, Sigma-Aldrich); tetanus toxoid (TT, Alomone Labs); recombinant influenza A virus H5N1 hemagglutinin (H5N1, Sino Biological Inc.); a recombinant immunoglobulin (Ig) light κ chain variable region (Vκ3–20 , a kind gift from Prof. R. Dolcetti, Centro di Riferimento Oncologico di Aviano (CRO-IRCCS), Aviano, Italy); and rabies virus G glycoprotein (RABV G; obtained from Challenge Virus Strain—11 (ATCC® VR-959™), propagated in BHK-21 cells (ATCC® CCL-10™), and purified as previously described ), were conjugated to HA-acetal using a general procedure for each antigen. ..

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    Sino Biological recombinant sars cov 2 spike protein
    Dose-dependent transduction rates of <t>SARS-CoV-2</t> pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.
    Recombinant Sars Cov 2 Spike Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Sino Biological h7n9
    Detection limit of FICT assay for target antigen. FICT employing Europium-conjugated antibodies were tested for the limit of detection (LOD) against <t>H7N9</t> rHA1 ( A ). The data ( n = 3) are shown as mean ± SD. Linear regression is shown with the dotted line. The arrow indicates the antigen concentration of LOD. Raw fluorescence peaks from the test line (TL) and control line (CL) in FICT are shown in the bottom panel.
    H7n9, supplied by Sino Biological, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Sino Biological pr8 h1n1 influenza virus strain
    CD4 and CD8 T cells are required for protective immunity to influenza A virus. C57BL/6 mice were vaccinated twice (at 3 weeks interval) with NP protein formulated in ADJ+GLA. At 70 days after booster vaccination, mice were challenged intranasally with <t>H1N1/PR8</t> strain of influenza A virus; unvaccinated mice were challenged as controls. Cohorts of vaccinated virus-challenged mice were treated (intravenously and intranasally) with isotype control IgG, anti-CD4 or anti-CD8 antibodies at days -5, -3, -1 and 1, 3 and 5, relative to viral challenge. On the 6 th day after viral challenge, virus-specific T cells and viral titers were quantified in lungs. (A) FACS plots are gated on live lymphocytes and numbers are percentages among live lymphocytes. (B) FACS plots are gated on CD8 T cells and numbers are percentages of D b /NP366 tetramer-binding CD8 T cells among CD8 T cells. (C) FACS plots are gated on CD4 T cells and numbers are percentages of I-A b /NP311 tetramer-binding CD4 T cells among CD8 T cells. (D) Viral titers in lungs were quantified by a plaque assay. Data are from two independent experiments. *, **, and *** indicate significance at P
    Pr8 H1n1 Influenza Virus Strain, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Sino Biological sars cov 2
    Dose-dependent transduction rates of <t>SARS-CoV-2</t> pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.
    Sars Cov 2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Dose-dependent transduction rates of SARS-CoV-2 pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Dose-dependent transduction rates of SARS-CoV-2 pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.

    Article Snippet: Mice immunizationFor SARS-CoV-2 antisera, 6–8 weeks old BALB/c mice were subcutaneously immunized twice with 50 μg of recombinant SARS-CoV-2 spike protein (S1+S2 ECD) (Sino Biological) and emulsified in Complete Freund's Adjuvant (CFA, Sigma) for priming and Incomplete Freund's Adjuvant (IFA, Sigma) for the boost in a total of 100 μl at a 3-week interval.

    Techniques: Transduction, Generated

    Lentiviral pseudovirus system of SARS-CoV or SARS-CoV-2 and avian influenza H5. Structural protein genes, including S protein of SARS-CoV or SARS-CoV-2 and HA/NA protein of avian influenza H5, were subcloned into envelope expression plasmid derived from pMD.G vector. To generate SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses, we co-transfected the structural protein expressing either S protein or HA and NA vectors, a package vector, and a reporter vector into HEK-293T cells. Generated SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses were harvested and transduced into Vero-E6 or MDCK cells, respectively.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Lentiviral pseudovirus system of SARS-CoV or SARS-CoV-2 and avian influenza H5. Structural protein genes, including S protein of SARS-CoV or SARS-CoV-2 and HA/NA protein of avian influenza H5, were subcloned into envelope expression plasmid derived from pMD.G vector. To generate SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses, we co-transfected the structural protein expressing either S protein or HA and NA vectors, a package vector, and a reporter vector into HEK-293T cells. Generated SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses were harvested and transduced into Vero-E6 or MDCK cells, respectively.

    Article Snippet: Mice immunizationFor SARS-CoV-2 antisera, 6–8 weeks old BALB/c mice were subcutaneously immunized twice with 50 μg of recombinant SARS-CoV-2 spike protein (S1+S2 ECD) (Sino Biological) and emulsified in Complete Freund's Adjuvant (CFA, Sigma) for priming and Incomplete Freund's Adjuvant (IFA, Sigma) for the boost in a total of 100 μl at a 3-week interval.

    Techniques: Expressing, Plasmid Preparation, Derivative Assay, Transfection, Generated

    Immunoblotting of S protein of SARS-CoV or SARS-CoV-2 and HA protein of avian influenza H5. (A) S proteins of SARS-CoV and SARS-CoV-2 were immunoblotted with mouse anti-SARS-CoV S protein antibody and mouse anti-HA tag protein antibody, respectively. (B) HA proteins of avian influenza H5 were immunoblotted with mouse anti-influenza virus H5 HA protein antibody. As the antibody recognized the HA2 epitope, both of HA0 and HA2 protein were detected by the immunoblotting.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Immunoblotting of S protein of SARS-CoV or SARS-CoV-2 and HA protein of avian influenza H5. (A) S proteins of SARS-CoV and SARS-CoV-2 were immunoblotted with mouse anti-SARS-CoV S protein antibody and mouse anti-HA tag protein antibody, respectively. (B) HA proteins of avian influenza H5 were immunoblotted with mouse anti-influenza virus H5 HA protein antibody. As the antibody recognized the HA2 epitope, both of HA0 and HA2 protein were detected by the immunoblotting.

    Article Snippet: Mice immunizationFor SARS-CoV-2 antisera, 6–8 weeks old BALB/c mice were subcutaneously immunized twice with 50 μg of recombinant SARS-CoV-2 spike protein (S1+S2 ECD) (Sino Biological) and emulsified in Complete Freund's Adjuvant (CFA, Sigma) for priming and Incomplete Freund's Adjuvant (IFA, Sigma) for the boost in a total of 100 μl at a 3-week interval.

    Techniques:

    Transduction optimization of SARS-CoV and SARS-CoV-2 pseudoviruses. Generated SARS-CoV and SARS-CoV-2 pseudoviruses were transduced into Vero-E6 cells. Different transduction medium with (A) 2% FBS or (B) 2.5 μg/ml trypsin. Using transduction medium with 2% FBS showed higher transduction rate for SARS-CoV and SARS-CoV-2 pseudoviruses. Using transduction medium with 2.5 μg/ml trypsin obviously reduced transduction rate, especially for SARS-CoV pseudoviruses.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Transduction optimization of SARS-CoV and SARS-CoV-2 pseudoviruses. Generated SARS-CoV and SARS-CoV-2 pseudoviruses were transduced into Vero-E6 cells. Different transduction medium with (A) 2% FBS or (B) 2.5 μg/ml trypsin. Using transduction medium with 2% FBS showed higher transduction rate for SARS-CoV and SARS-CoV-2 pseudoviruses. Using transduction medium with 2.5 μg/ml trypsin obviously reduced transduction rate, especially for SARS-CoV pseudoviruses.

    Article Snippet: Mice immunizationFor SARS-CoV-2 antisera, 6–8 weeks old BALB/c mice were subcutaneously immunized twice with 50 μg of recombinant SARS-CoV-2 spike protein (S1+S2 ECD) (Sino Biological) and emulsified in Complete Freund's Adjuvant (CFA, Sigma) for priming and Incomplete Freund's Adjuvant (IFA, Sigma) for the boost in a total of 100 μl at a 3-week interval.

    Techniques: Transduction, Generated

    Pseudovirus transduction of SARS-CoV or SARS-CoV-2 and avian influenza H5Nx. Generated (A) SARS-CoV or SARS-CoV-2 and (B) avian influenza H5Nx pseudoviruses were transduced into Vero-E6 or MDCK cells, respectively. Red fluorescence indicated the cells transduced by the indicated pseudoviruses with RFP reporter gene. (C) Transduction titers of avian influenza H5Nx pseudoviruses were determined according to the numbers of cells expressing red fluorescence.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Pseudovirus transduction of SARS-CoV or SARS-CoV-2 and avian influenza H5Nx. Generated (A) SARS-CoV or SARS-CoV-2 and (B) avian influenza H5Nx pseudoviruses were transduced into Vero-E6 or MDCK cells, respectively. Red fluorescence indicated the cells transduced by the indicated pseudoviruses with RFP reporter gene. (C) Transduction titers of avian influenza H5Nx pseudoviruses were determined according to the numbers of cells expressing red fluorescence.

    Article Snippet: Mice immunizationFor SARS-CoV-2 antisera, 6–8 weeks old BALB/c mice were subcutaneously immunized twice with 50 μg of recombinant SARS-CoV-2 spike protein (S1+S2 ECD) (Sino Biological) and emulsified in Complete Freund's Adjuvant (CFA, Sigma) for priming and Incomplete Freund's Adjuvant (IFA, Sigma) for the boost in a total of 100 μl at a 3-week interval.

    Techniques: Transduction, Generated, Fluorescence, Expressing

    Detection limit of FICT assay for target antigen. FICT employing Europium-conjugated antibodies were tested for the limit of detection (LOD) against H7N9 rHA1 ( A ). The data ( n = 3) are shown as mean ± SD. Linear regression is shown with the dotted line. The arrow indicates the antigen concentration of LOD. Raw fluorescence peaks from the test line (TL) and control line (CL) in FICT are shown in the bottom panel.

    Journal: Scientific Reports

    Article Title: Improvement of a rapid diagnostic application of monoclonal antibodies against avian influenza H7 subtype virus using Europium nanoparticles

    doi: 10.1038/s41598-017-08328-9

    Figure Lengend Snippet: Detection limit of FICT assay for target antigen. FICT employing Europium-conjugated antibodies were tested for the limit of detection (LOD) against H7N9 rHA1 ( A ). The data ( n = 3) are shown as mean ± SD. Linear regression is shown with the dotted line. The arrow indicates the antigen concentration of LOD. Raw fluorescence peaks from the test line (TL) and control line (CL) in FICT are shown in the bottom panel.

    Article Snippet: Recombinant hemagglutinin 1 (rHA1) of H7N9 (A/Anhui/1/2013) and rHA1 of H5N1 (A/Vietnam/1/2003) were purchased from Sino Biological Inc. (Beijing, China).

    Techniques: Concentration Assay, Fluorescence

    Development of H7 subtype-specific antibodies. The fusions were performed using mouse spleen cells inoculated with H7N9 virus rHA1. Ten hybridomas were produced. The secreted antibodies from each hybridoma were tested for recombinant antigen (H7N9 HA1 ( A ) and different influenza subtype virus ( B ) by indirect ELISA. Pre-immune, serum from a healthy mouse; P.C., positive control antibody (anti-influenza A nucleoprotein antibody).

    Journal: Scientific Reports

    Article Title: Improvement of a rapid diagnostic application of monoclonal antibodies against avian influenza H7 subtype virus using Europium nanoparticles

    doi: 10.1038/s41598-017-08328-9

    Figure Lengend Snippet: Development of H7 subtype-specific antibodies. The fusions were performed using mouse spleen cells inoculated with H7N9 virus rHA1. Ten hybridomas were produced. The secreted antibodies from each hybridoma were tested for recombinant antigen (H7N9 HA1 ( A ) and different influenza subtype virus ( B ) by indirect ELISA. Pre-immune, serum from a healthy mouse; P.C., positive control antibody (anti-influenza A nucleoprotein antibody).

    Article Snippet: Recombinant hemagglutinin 1 (rHA1) of H7N9 (A/Anhui/1/2013) and rHA1 of H5N1 (A/Vietnam/1/2003) were purchased from Sino Biological Inc. (Beijing, China).

    Techniques: Produced, Recombinant, Indirect ELISA, Positive Control

    CD4 and CD8 T cells are required for protective immunity to influenza A virus. C57BL/6 mice were vaccinated twice (at 3 weeks interval) with NP protein formulated in ADJ+GLA. At 70 days after booster vaccination, mice were challenged intranasally with H1N1/PR8 strain of influenza A virus; unvaccinated mice were challenged as controls. Cohorts of vaccinated virus-challenged mice were treated (intravenously and intranasally) with isotype control IgG, anti-CD4 or anti-CD8 antibodies at days -5, -3, -1 and 1, 3 and 5, relative to viral challenge. On the 6 th day after viral challenge, virus-specific T cells and viral titers were quantified in lungs. (A) FACS plots are gated on live lymphocytes and numbers are percentages among live lymphocytes. (B) FACS plots are gated on CD8 T cells and numbers are percentages of D b /NP366 tetramer-binding CD8 T cells among CD8 T cells. (C) FACS plots are gated on CD4 T cells and numbers are percentages of I-A b /NP311 tetramer-binding CD4 T cells among CD8 T cells. (D) Viral titers in lungs were quantified by a plaque assay. Data are from two independent experiments. *, **, and *** indicate significance at P

    Journal: bioRxiv

    Article Title: Programming Multifaceted Pulmonary T-Cell Immunity by Combination Adjuvants

    doi: 10.1101/2020.07.10.197459

    Figure Lengend Snippet: CD4 and CD8 T cells are required for protective immunity to influenza A virus. C57BL/6 mice were vaccinated twice (at 3 weeks interval) with NP protein formulated in ADJ+GLA. At 70 days after booster vaccination, mice were challenged intranasally with H1N1/PR8 strain of influenza A virus; unvaccinated mice were challenged as controls. Cohorts of vaccinated virus-challenged mice were treated (intravenously and intranasally) with isotype control IgG, anti-CD4 or anti-CD8 antibodies at days -5, -3, -1 and 1, 3 and 5, relative to viral challenge. On the 6 th day after viral challenge, virus-specific T cells and viral titers were quantified in lungs. (A) FACS plots are gated on live lymphocytes and numbers are percentages among live lymphocytes. (B) FACS plots are gated on CD8 T cells and numbers are percentages of D b /NP366 tetramer-binding CD8 T cells among CD8 T cells. (C) FACS plots are gated on CD4 T cells and numbers are percentages of I-A b /NP311 tetramer-binding CD4 T cells among CD8 T cells. (D) Viral titers in lungs were quantified by a plaque assay. Data are from two independent experiments. *, **, and *** indicate significance at P

    Article Snippet: Recombinant nucleoprotein (NP) of the PR8/H1N1 influenza virus strain was purchased from Sino Biological Inc (Beijing, China).

    Techniques: Mouse Assay, FACS, Binding Assay, Plaque Assay

    Histopathological analysis of lungs following viral challenge of vaccinated mice. Groups of C57BL/6 mice were vaccinated twice (at 3 weeks interval) with NP protein formulated in various adjuvants. At 100 days after booster vaccination, vaccinated mice were challenged intranasally with H1N1/PR8 strain of influenza A virus. On the 6 th day after viral challenge, lungs were collected in neutral-buffered formailin, and tissue sections were stained with Hematoxylin and Eosin (H E). H E stained lung sections were evaluated by a board-certified pathologist (Dr. Gasper); he was blinded to the identity of sections. In each image (40X magnification), asterisks indicate similarly sized large bronchioles, arrow heads indicate regions in which bronchial lesions extend in to the adjacent alveoli, and arrows indicate perivascular lymphoid nodules. A. Adjuplex-vaccinated mouse: there is mild necrotizing bronchitis asterisks). B. CPG-vaccinated mouse: there is obliteration of two bronchioles by inflammation that extends far into the surrounding alveoli (arrowheads). C. GLA-vaccinated mouse: there is bronchiolitis affecting 1 of the larger bronchioles, with minimal extension into the adjacent alveoli. D. ADJ+CPG-vaccinated mouse. Broncholitis is similar to that in A, but alveolar regions around the affected bronchiole (center) are infiltrated by inflammatory cells. E. ADJ+GLA vaccinated mouse: bronchiolitis is of intermediate severity between B and C, and regionally extends into the adjacent alveolar tissue (arrowhead). Each lung section was scored individually, and lesion scores from 0-3 were assigned for bronchial lesions, alveolar lesions, and specific disease patterns, with 0 = absent, 1 = mild, 2 = moderate, 3 = severe. Bronchioloar Lesions: Epithelial degeneration/necrosis; Intraepithelial neutrophils; Intraepithelial eosinophils; Intraepithelial lymphocytes; Luminal dislodged epithelial cells/debris; Luminal cellular exudate; Peribronchiolar neutrophils; Pavementing/Subendothelial leukocytes. Alveolar Lesions: Alveolar wall thickening; Interstitial macrophages; Interstitial lymphocytes; Interstitial granulocytes; Epithelial necrosis; Luminal edema; Luminal hemorrhage; Luminal cellular exudate; Luminal alveolar macrophages; Luminal neutrophils; Luminal sloughed epithelial cells.

    Journal: bioRxiv

    Article Title: Programming Multifaceted Pulmonary T-Cell Immunity by Combination Adjuvants

    doi: 10.1101/2020.07.10.197459

    Figure Lengend Snippet: Histopathological analysis of lungs following viral challenge of vaccinated mice. Groups of C57BL/6 mice were vaccinated twice (at 3 weeks interval) with NP protein formulated in various adjuvants. At 100 days after booster vaccination, vaccinated mice were challenged intranasally with H1N1/PR8 strain of influenza A virus. On the 6 th day after viral challenge, lungs were collected in neutral-buffered formailin, and tissue sections were stained with Hematoxylin and Eosin (H E). H E stained lung sections were evaluated by a board-certified pathologist (Dr. Gasper); he was blinded to the identity of sections. In each image (40X magnification), asterisks indicate similarly sized large bronchioles, arrow heads indicate regions in which bronchial lesions extend in to the adjacent alveoli, and arrows indicate perivascular lymphoid nodules. A. Adjuplex-vaccinated mouse: there is mild necrotizing bronchitis asterisks). B. CPG-vaccinated mouse: there is obliteration of two bronchioles by inflammation that extends far into the surrounding alveoli (arrowheads). C. GLA-vaccinated mouse: there is bronchiolitis affecting 1 of the larger bronchioles, with minimal extension into the adjacent alveoli. D. ADJ+CPG-vaccinated mouse. Broncholitis is similar to that in A, but alveolar regions around the affected bronchiole (center) are infiltrated by inflammatory cells. E. ADJ+GLA vaccinated mouse: bronchiolitis is of intermediate severity between B and C, and regionally extends into the adjacent alveolar tissue (arrowhead). Each lung section was scored individually, and lesion scores from 0-3 were assigned for bronchial lesions, alveolar lesions, and specific disease patterns, with 0 = absent, 1 = mild, 2 = moderate, 3 = severe. Bronchioloar Lesions: Epithelial degeneration/necrosis; Intraepithelial neutrophils; Intraepithelial eosinophils; Intraepithelial lymphocytes; Luminal dislodged epithelial cells/debris; Luminal cellular exudate; Peribronchiolar neutrophils; Pavementing/Subendothelial leukocytes. Alveolar Lesions: Alveolar wall thickening; Interstitial macrophages; Interstitial lymphocytes; Interstitial granulocytes; Epithelial necrosis; Luminal edema; Luminal hemorrhage; Luminal cellular exudate; Luminal alveolar macrophages; Luminal neutrophils; Luminal sloughed epithelial cells.

    Article Snippet: Recombinant nucleoprotein (NP) of the PR8/H1N1 influenza virus strain was purchased from Sino Biological Inc (Beijing, China).

    Techniques: Mouse Assay, Staining

    Vaccine-induced protective immunity to H1N1 and H5N1 influenza viruses. (A-C) Groups of C57BL/6 mice were vaccinated twice at 3-week intervals with NP protein formulated in various adjuvants.. (A) At 100 days after the booster vaccination, mice were challenged intranasally with H1N1/PR8 strain of influenza A virus. Viral tiers and virus-specific T cell responses in lungs were quantified on the 6 th day after virus challenge. (A) Viral titers in the lungs on the 6 th day after virus challenge. (B) Percentages of NP366-specific IFN-γ and IL-17 producing cells among CD8 T cells (bar graphs) and calculated proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ+IL-7-producing peptide-stimulated NP366-specific T cells (Pie charts). (C) Percentages of NP311-specific IFN-γ and IL-17 producing cells among CD4 T cells and calculated proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ+IL-7-producing peptide-stimulated NP311-specific T cells. (D) C57BL/6 mice were vaccinated with NP+ADJ+GLA twice at an interval of 3 weeks. One hundred and eighty days after the last vaccination, mice were challenged intransally with H1N1/PR8 strain of influenza A virus; unvaccinated mice were challenged with virus as controls. Cohorts of vaccinated virus-challenged mice were treated with isotype control IgG or anti-IL-17A antibodies (intravenously and intranasally) at -1, 0, 1, 3 and 5 days relative to virus challenge. On the 6 th day after viral challenge, viral titers and virus-specific T cell responses were quantified in lungs. (E) Groups of C57BL/6 mice were vaccinated twice with NP protein alone or formulated in various adjuvants. Fifty days after booster vaccination, vaccinated and unvaccinated mice were challenged intranasally with the highly pathogenic H5N1 avian influenza A virus; weight loss and survival was monitored until day 14. Data are pooled from 2 independent experiments or representative of two independent experiments. *, **, and *** indicate significance at P

    Journal: bioRxiv

    Article Title: Programming Multifaceted Pulmonary T-Cell Immunity by Combination Adjuvants

    doi: 10.1101/2020.07.10.197459

    Figure Lengend Snippet: Vaccine-induced protective immunity to H1N1 and H5N1 influenza viruses. (A-C) Groups of C57BL/6 mice were vaccinated twice at 3-week intervals with NP protein formulated in various adjuvants.. (A) At 100 days after the booster vaccination, mice were challenged intranasally with H1N1/PR8 strain of influenza A virus. Viral tiers and virus-specific T cell responses in lungs were quantified on the 6 th day after virus challenge. (A) Viral titers in the lungs on the 6 th day after virus challenge. (B) Percentages of NP366-specific IFN-γ and IL-17 producing cells among CD8 T cells (bar graphs) and calculated proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ+IL-7-producing peptide-stimulated NP366-specific T cells (Pie charts). (C) Percentages of NP311-specific IFN-γ and IL-17 producing cells among CD4 T cells and calculated proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ+IL-7-producing peptide-stimulated NP311-specific T cells. (D) C57BL/6 mice were vaccinated with NP+ADJ+GLA twice at an interval of 3 weeks. One hundred and eighty days after the last vaccination, mice were challenged intransally with H1N1/PR8 strain of influenza A virus; unvaccinated mice were challenged with virus as controls. Cohorts of vaccinated virus-challenged mice were treated with isotype control IgG or anti-IL-17A antibodies (intravenously and intranasally) at -1, 0, 1, 3 and 5 days relative to virus challenge. On the 6 th day after viral challenge, viral titers and virus-specific T cell responses were quantified in lungs. (E) Groups of C57BL/6 mice were vaccinated twice with NP protein alone or formulated in various adjuvants. Fifty days after booster vaccination, vaccinated and unvaccinated mice were challenged intranasally with the highly pathogenic H5N1 avian influenza A virus; weight loss and survival was monitored until day 14. Data are pooled from 2 independent experiments or representative of two independent experiments. *, **, and *** indicate significance at P

    Article Snippet: Recombinant nucleoprotein (NP) of the PR8/H1N1 influenza virus strain was purchased from Sino Biological Inc (Beijing, China).

    Techniques: Mouse Assay

    Kinetics and durability of influenza viral control in vaccinated mice. B6 mice were vaccinated twice (at 3 weeks intervals) intranasally with NP protein formulated with the indicated adjuvants. Unvaccinated mice and mice vaccinated with NP only (without adjuvants) served as controls. At 100 and 180 days after booster vaccination, mice were challenged intranasally with PR8/H1N1 influenza virus. (A) Body weight loss was assessed by calculating bodyweight at different days after challenge, relative to bodyweight before challenge at 100 days after vaccination. (B) Vaccinated mice were challenged with PR8/H1N1at 100 days after vaccination and viral titers in lungs were quantified at day 2 and 4 after challenge, using a plaque assay. (C) At day 180 after booster vaccination, mice were challenged with PR8/H1N1 virus, and viral titers in lungs were assessed at day 6 after challenge. (D) Percentages and numbers of NP366-specific CD8 T cells and NP311-specific CD4 T cells in lungs and percentage of these cells in the vascular and non-vascular compartment at day 6 after challenge (challenged at 100 days after vaccination). *, **, and *** indicate significance at P

    Journal: bioRxiv

    Article Title: Programming Multifaceted Pulmonary T-Cell Immunity by Combination Adjuvants

    doi: 10.1101/2020.07.10.197459

    Figure Lengend Snippet: Kinetics and durability of influenza viral control in vaccinated mice. B6 mice were vaccinated twice (at 3 weeks intervals) intranasally with NP protein formulated with the indicated adjuvants. Unvaccinated mice and mice vaccinated with NP only (without adjuvants) served as controls. At 100 and 180 days after booster vaccination, mice were challenged intranasally with PR8/H1N1 influenza virus. (A) Body weight loss was assessed by calculating bodyweight at different days after challenge, relative to bodyweight before challenge at 100 days after vaccination. (B) Vaccinated mice were challenged with PR8/H1N1at 100 days after vaccination and viral titers in lungs were quantified at day 2 and 4 after challenge, using a plaque assay. (C) At day 180 after booster vaccination, mice were challenged with PR8/H1N1 virus, and viral titers in lungs were assessed at day 6 after challenge. (D) Percentages and numbers of NP366-specific CD8 T cells and NP311-specific CD4 T cells in lungs and percentage of these cells in the vascular and non-vascular compartment at day 6 after challenge (challenged at 100 days after vaccination). *, **, and *** indicate significance at P

    Article Snippet: Recombinant nucleoprotein (NP) of the PR8/H1N1 influenza virus strain was purchased from Sino Biological Inc (Beijing, China).

    Techniques: Mouse Assay, Plaque Assay

    Regulation of vaccine-induced CD8 T-cell memory and protective immunity by CD4 T cells. Groups of C57BL/6 mice were vaccinated twice at 3-week intervals with NP protein formulated in ADJ+GLA. Cohorts of vaccinated mice were treated with isotype control antibodies (Non Depleted) or anti-CD4 antibodies (CD4 Depleted) intravenously and intranasally on days −1, 0, and 1 relative to prime and boost vaccination with NP+ADJ+GLA. T-cell memory in lungs (A-F) and protective immunity to influenza A virus (G-P) was determined at 80 days after booster vaccination. (A-F) T-cell memory in lungs at day 80 after booster vaccination. To stain for vascular cells, mice were injected intravenously with anti-CD45.2 antibodies, 3 minutes prior to euthanasia. Lung cells were stained directly ex vivo with D b /NP366 or I-A b /NP311 tetramers along with the indicated antibodies for cell surface markers. For cytokine analysis, lung cells were stimulated with NP366 or NP311 peptide for 5 hours before intracellular staining. (A) FACS plots are gated on total CD4 T cells and show NP311-specific tetramer-binding memory CD4 T cells only in non-depleted mice. (B) NP366-specific tetramer-binding memory CD8 T cells in lungs of non-d epleted and CD4 T cell-depleted mice. (C) Expression of tissue residency markers on NP366-specific tetramer-binding memory CD8 T cells in lungs. (D) Percentage of vascular (CD45.2 +ve ) and non-vascular (CD45.2 -ve ) cells among NP366-specific tetramer-binding memory CD8 T cells in lungs. (E) Percentages of IFN-γ- and IL-17-producing NP366-specific cells among CD8 T cells in lungs. (F) Calculated proportions of IFN-γ and/or IL-17-producing cells among cytokine-producing peptide-stimulated IFN-γ+IL-17 NP366-specific CD8 T cells. (G-P) At day 80 after booster vaccination, non-depleted and CD4 T cell-depleted mice were challenged intranasally with PR8/H1N1 influenza A virus; recall virus-specific CD8/CD4 T cell responses and viral load in lungs were assessed at day 6 after challenge. (G) Percentages of NP366-specific tetramer-binding cells among CD8 T cells in lungs. (H) Percentages of NP366-specific tetramer-binding CD8 T cells in vascular and nonvascular lung compartment. (I) Percentages of NP311-specific tetramer-binding cells among CD4 T cells in lungs. (J) Expression of tissue residency markers on NP366-specific tetramer-binding CD8 T cells. (K) Chemokine receptor and transcription factor expression in NP366-specific CD8 T cells in lungs. (L) Granzyme B expression by NP366-specific CD8 T cells directly ex vivo. (M) Percentages of IFN-γ and IL-17 producing NP366-specific CD8 T cells. (N) Relative proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ plus IL-17-producing peptide-stimulated NP366-specific CD8 T cells. (O) Viral titers in lungs at day 6 after challenge. (P) Body weight, measured as a percentage of starting body weight prior to challenge. Data are pooled from two independent experiments. *, **, and *** indicate significance at P

    Journal: bioRxiv

    Article Title: Programming Multifaceted Pulmonary T-Cell Immunity by Combination Adjuvants

    doi: 10.1101/2020.07.10.197459

    Figure Lengend Snippet: Regulation of vaccine-induced CD8 T-cell memory and protective immunity by CD4 T cells. Groups of C57BL/6 mice were vaccinated twice at 3-week intervals with NP protein formulated in ADJ+GLA. Cohorts of vaccinated mice were treated with isotype control antibodies (Non Depleted) or anti-CD4 antibodies (CD4 Depleted) intravenously and intranasally on days −1, 0, and 1 relative to prime and boost vaccination with NP+ADJ+GLA. T-cell memory in lungs (A-F) and protective immunity to influenza A virus (G-P) was determined at 80 days after booster vaccination. (A-F) T-cell memory in lungs at day 80 after booster vaccination. To stain for vascular cells, mice were injected intravenously with anti-CD45.2 antibodies, 3 minutes prior to euthanasia. Lung cells were stained directly ex vivo with D b /NP366 or I-A b /NP311 tetramers along with the indicated antibodies for cell surface markers. For cytokine analysis, lung cells were stimulated with NP366 or NP311 peptide for 5 hours before intracellular staining. (A) FACS plots are gated on total CD4 T cells and show NP311-specific tetramer-binding memory CD4 T cells only in non-depleted mice. (B) NP366-specific tetramer-binding memory CD8 T cells in lungs of non-d epleted and CD4 T cell-depleted mice. (C) Expression of tissue residency markers on NP366-specific tetramer-binding memory CD8 T cells in lungs. (D) Percentage of vascular (CD45.2 +ve ) and non-vascular (CD45.2 -ve ) cells among NP366-specific tetramer-binding memory CD8 T cells in lungs. (E) Percentages of IFN-γ- and IL-17-producing NP366-specific cells among CD8 T cells in lungs. (F) Calculated proportions of IFN-γ and/or IL-17-producing cells among cytokine-producing peptide-stimulated IFN-γ+IL-17 NP366-specific CD8 T cells. (G-P) At day 80 after booster vaccination, non-depleted and CD4 T cell-depleted mice were challenged intranasally with PR8/H1N1 influenza A virus; recall virus-specific CD8/CD4 T cell responses and viral load in lungs were assessed at day 6 after challenge. (G) Percentages of NP366-specific tetramer-binding cells among CD8 T cells in lungs. (H) Percentages of NP366-specific tetramer-binding CD8 T cells in vascular and nonvascular lung compartment. (I) Percentages of NP311-specific tetramer-binding cells among CD4 T cells in lungs. (J) Expression of tissue residency markers on NP366-specific tetramer-binding CD8 T cells. (K) Chemokine receptor and transcription factor expression in NP366-specific CD8 T cells in lungs. (L) Granzyme B expression by NP366-specific CD8 T cells directly ex vivo. (M) Percentages of IFN-γ and IL-17 producing NP366-specific CD8 T cells. (N) Relative proportions of IFN-γ and/or IL-17 producing cells among total IFN-γ plus IL-17-producing peptide-stimulated NP366-specific CD8 T cells. (O) Viral titers in lungs at day 6 after challenge. (P) Body weight, measured as a percentage of starting body weight prior to challenge. Data are pooled from two independent experiments. *, **, and *** indicate significance at P

    Article Snippet: Recombinant nucleoprotein (NP) of the PR8/H1N1 influenza virus strain was purchased from Sino Biological Inc (Beijing, China).

    Techniques: Mouse Assay, Staining, Injection, Ex Vivo, FACS, Binding Assay, Expressing

    Dose-dependent transduction rates of SARS-CoV-2 pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Dose-dependent transduction rates of SARS-CoV-2 pseudoviruses. Generated SARS-CoV-2 pseudoviruses were serially diluted and then transduced into Vero-E6 cells. Transduction rate of SARS-CoV-2 was gradually reduced in a dose-dependent manner. According to the transduction rate curve, the titer of SARS-CoV-2 pseudovirus was quantified as 2.33 × 10 5 transduction unit.

    Article Snippet: Production of SARS-CoV/SARS-CoV-2 and avian influenza viruses H5Nx pseudoviruses To generate SARS-CoV or SARS-CoV-2 and avian influenza virus pseudovirus, we applied the lentiviral vector system provided by National RNAi Core of Academic Sinica Taiwan to produce the pseudoviruses expressing full-length S protein and HA/NA proteins, respectively.

    Techniques: Transduction, Generated

    Lentiviral pseudovirus system of SARS-CoV or SARS-CoV-2 and avian influenza H5. Structural protein genes, including S protein of SARS-CoV or SARS-CoV-2 and HA/NA protein of avian influenza H5, were subcloned into envelope expression plasmid derived from pMD.G vector. To generate SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses, we co-transfected the structural protein expressing either S protein or HA and NA vectors, a package vector, and a reporter vector into HEK-293T cells. Generated SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses were harvested and transduced into Vero-E6 or MDCK cells, respectively.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Lentiviral pseudovirus system of SARS-CoV or SARS-CoV-2 and avian influenza H5. Structural protein genes, including S protein of SARS-CoV or SARS-CoV-2 and HA/NA protein of avian influenza H5, were subcloned into envelope expression plasmid derived from pMD.G vector. To generate SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses, we co-transfected the structural protein expressing either S protein or HA and NA vectors, a package vector, and a reporter vector into HEK-293T cells. Generated SARS-CoV or SARS-CoV-2 and avian influenza H5Nx pseudoviruses were harvested and transduced into Vero-E6 or MDCK cells, respectively.

    Article Snippet: Production of SARS-CoV/SARS-CoV-2 and avian influenza viruses H5Nx pseudoviruses To generate SARS-CoV or SARS-CoV-2 and avian influenza virus pseudovirus, we applied the lentiviral vector system provided by National RNAi Core of Academic Sinica Taiwan to produce the pseudoviruses expressing full-length S protein and HA/NA proteins, respectively.

    Techniques: Expressing, Plasmid Preparation, Derivative Assay, Transfection, Generated

    Immunoblotting of S protein of SARS-CoV or SARS-CoV-2 and HA protein of avian influenza H5. (A) S proteins of SARS-CoV and SARS-CoV-2 were immunoblotted with mouse anti-SARS-CoV S protein antibody and mouse anti-HA tag protein antibody, respectively. (B) HA proteins of avian influenza H5 were immunoblotted with mouse anti-influenza virus H5 HA protein antibody. As the antibody recognized the HA2 epitope, both of HA0 and HA2 protein were detected by the immunoblotting.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Immunoblotting of S protein of SARS-CoV or SARS-CoV-2 and HA protein of avian influenza H5. (A) S proteins of SARS-CoV and SARS-CoV-2 were immunoblotted with mouse anti-SARS-CoV S protein antibody and mouse anti-HA tag protein antibody, respectively. (B) HA proteins of avian influenza H5 were immunoblotted with mouse anti-influenza virus H5 HA protein antibody. As the antibody recognized the HA2 epitope, both of HA0 and HA2 protein were detected by the immunoblotting.

    Article Snippet: Production of SARS-CoV/SARS-CoV-2 and avian influenza viruses H5Nx pseudoviruses To generate SARS-CoV or SARS-CoV-2 and avian influenza virus pseudovirus, we applied the lentiviral vector system provided by National RNAi Core of Academic Sinica Taiwan to produce the pseudoviruses expressing full-length S protein and HA/NA proteins, respectively.

    Techniques:

    Transduction optimization of SARS-CoV and SARS-CoV-2 pseudoviruses. Generated SARS-CoV and SARS-CoV-2 pseudoviruses were transduced into Vero-E6 cells. Different transduction medium with (A) 2% FBS or (B) 2.5 μg/ml trypsin. Using transduction medium with 2% FBS showed higher transduction rate for SARS-CoV and SARS-CoV-2 pseudoviruses. Using transduction medium with 2.5 μg/ml trypsin obviously reduced transduction rate, especially for SARS-CoV pseudoviruses.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Transduction optimization of SARS-CoV and SARS-CoV-2 pseudoviruses. Generated SARS-CoV and SARS-CoV-2 pseudoviruses were transduced into Vero-E6 cells. Different transduction medium with (A) 2% FBS or (B) 2.5 μg/ml trypsin. Using transduction medium with 2% FBS showed higher transduction rate for SARS-CoV and SARS-CoV-2 pseudoviruses. Using transduction medium with 2.5 μg/ml trypsin obviously reduced transduction rate, especially for SARS-CoV pseudoviruses.

    Article Snippet: Production of SARS-CoV/SARS-CoV-2 and avian influenza viruses H5Nx pseudoviruses To generate SARS-CoV or SARS-CoV-2 and avian influenza virus pseudovirus, we applied the lentiviral vector system provided by National RNAi Core of Academic Sinica Taiwan to produce the pseudoviruses expressing full-length S protein and HA/NA proteins, respectively.

    Techniques: Transduction, Generated

    Pseudovirus transduction of SARS-CoV or SARS-CoV-2 and avian influenza H5Nx. Generated (A) SARS-CoV or SARS-CoV-2 and (B) avian influenza H5Nx pseudoviruses were transduced into Vero-E6 or MDCK cells, respectively. Red fluorescence indicated the cells transduced by the indicated pseudoviruses with RFP reporter gene. (C) Transduction titers of avian influenza H5Nx pseudoviruses were determined according to the numbers of cells expressing red fluorescence.

    Journal: Biomedical Journal

    Article Title: Assessing the application of a pseudovirus system for emerging SARS-CoV-2 and re-emerging avian influenza virus H5 subtypes in vaccine development

    doi: 10.1016/j.bj.2020.06.003

    Figure Lengend Snippet: Pseudovirus transduction of SARS-CoV or SARS-CoV-2 and avian influenza H5Nx. Generated (A) SARS-CoV or SARS-CoV-2 and (B) avian influenza H5Nx pseudoviruses were transduced into Vero-E6 or MDCK cells, respectively. Red fluorescence indicated the cells transduced by the indicated pseudoviruses with RFP reporter gene. (C) Transduction titers of avian influenza H5Nx pseudoviruses were determined according to the numbers of cells expressing red fluorescence.

    Article Snippet: Production of SARS-CoV/SARS-CoV-2 and avian influenza viruses H5Nx pseudoviruses To generate SARS-CoV or SARS-CoV-2 and avian influenza virus pseudovirus, we applied the lentiviral vector system provided by National RNAi Core of Academic Sinica Taiwan to produce the pseudoviruses expressing full-length S protein and HA/NA proteins, respectively.

    Techniques: Transduction, Generated, Fluorescence, Expressing