human rsv  (ATCC)


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    Structured Review

    ATCC human rsv
    Human Rsv, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human rsv/product/ATCC
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    human rsv - by Bioz Stars, 2020-01
    93/100 stars

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    Related Articles

    Histopathology:

    Article Title: An immunocompromised BALB/c mouse model for respiratory syncytial virus infection
    Article Snippet: RSV infection, weight determination and tissue collection The A2 strain of human RSV (American Type Culture Collection, Manassas, VA) was propagated in Hep-2 cells (ATCC) in a monolayer culture as previously described (Behera et al., 1998). .. A second set of animals was sacrificed five days after infection and their lungs were removed for determination of RSV titers, cytokine levels and histopathology.

    Infection:

    Article Title: An immunocompromised BALB/c mouse model for respiratory syncytial virus infection
    Article Snippet: .. RSV infection, weight determination and tissue collection The A2 strain of human RSV (American Type Culture Collection, Manassas, VA) was propagated in Hep-2 cells (ATCC) in a monolayer culture as previously described (Behera et al., 1998). .. Mice were infected intranasally with 5 × 105 PFU of RSV in a volume of 50 μl five days after treatment with CYP.

    Article Title: Combined fluticasone propionate and salmeterol reduces RSV infection more effectively than either of them alone in allergen-sensitized mice
    Article Snippet: .. RSV preparation and infection of mice The A2 strain of human RSV (American Type Culture Collection, Manassas, VA) was propagated in HEp-2 cells (American Type Culture Collection) grown in Eagle's minimal essential medium (Gibco) with 2% FBS. ..

    Article Title: MMP-12-mediated by SARM-TRIF signaling pathway contributes to IFN-γ-independent airway inflammation and AHR post RSV infection in nude mice
    Article Snippet: RSV preparation and mice treatment We utilized the A2 strain of human RSV (American Type Culture Collection). .. To assess the effect of MMP-12 on the airway inflammation and AHR, both BALB/c mice and nude mice were treated with MMP408, a potent and specific MMP-12 inhibitor, (CALBIOCHEM, EMD Chemicals, Inc. San Diego, CA 92121) at 5 mg/kg or PBS intragastrically twice a day from day 0 to day 8 post infection.

    Cell Culture:

    Article Title: MMP-12-mediated by SARM-TRIF signaling pathway contributes to IFN-γ-independent airway inflammation and AHR post RSV infection in nude mice
    Article Snippet: RSV preparation and mice treatment We utilized the A2 strain of human RSV (American Type Culture Collection). .. To inoculate RSV, the mice were held upright after sedation and inoculated intranasally with 1.5 × 107 PFU RSV in a 100-μl volume or sham-infected with 100-μl cell culture supernatant (mock group).

    Mouse Assay:

    Article Title: An immunocompromised BALB/c mouse model for respiratory syncytial virus infection
    Article Snippet: RSV infection, weight determination and tissue collection The A2 strain of human RSV (American Type Culture Collection, Manassas, VA) was propagated in Hep-2 cells (ATCC) in a monolayer culture as previously described (Behera et al., 1998). .. Mice were infected intranasally with 5 × 105 PFU of RSV in a volume of 50 μl five days after treatment with CYP.

    Article Title: Combined fluticasone propionate and salmeterol reduces RSV infection more effectively than either of them alone in allergen-sensitized mice
    Article Snippet: .. RSV preparation and infection of mice The A2 strain of human RSV (American Type Culture Collection, Manassas, VA) was propagated in HEp-2 cells (American Type Culture Collection) grown in Eagle's minimal essential medium (Gibco) with 2% FBS. ..

    Article Title: MMP-12-mediated by SARM-TRIF signaling pathway contributes to IFN-γ-independent airway inflammation and AHR post RSV infection in nude mice
    Article Snippet: .. RSV preparation and mice treatment We utilized the A2 strain of human RSV (American Type Culture Collection). .. To inoculate RSV, the mice were held upright after sedation and inoculated intranasally with 1.5 × 107 PFU RSV in a 100-μl volume or sham-infected with 100-μl cell culture supernatant (mock group).

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    ATCC rsv infected human epithelial cell lines
    NLRP3 and ASC are required for caspase-1 activation following <t>RSV</t> infection. ( A ) Wild type (WT), NLRP3 knock-out (KO) and ASC KO BMDMs were infected with RSV (1 MOI) for 12 h. The cell lysate from mock infected and RSV infected cells were subjected to Western blot analysis with mouse caspase-1 p10 subunit specific antibody. ( B ) RT-PCR analysis of <t>pro-IL-1β,</t> ASC and NLRP3 expression in RSV infected WT, NLRP3 KO and ASC KO BMDMs. The gels shown in (A) and (B) is representative of three independent experiments that yielded similar results.
    Rsv Infected Human Epithelial Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rsv infected human epithelial cell lines/product/ATCC
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    rsv infected human epithelial cell lines - by Bioz Stars, 2020-01
    78/100 stars
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    92
    ATCC human rsv a2
    Co-infection elicited reduction in the number of CD11b or CD11c myeloid cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), <t>RSV</t> A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. to determine the frequency of NK1.1 gated on CD3 negative cells. (A) Proportion of CD11b myeloid cells, which was significantly low in the I + R group on day 2 was increased by day 7. (B) Proportions of CD11c myeloid cells were lower in I + R group compared to other groups on days 2 and 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.
    Human Rsv A2, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human rsv a2/product/ATCC
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    human rsv a2 - by Bioz Stars, 2020-01
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    NLRP3 and ASC are required for caspase-1 activation following RSV infection. ( A ) Wild type (WT), NLRP3 knock-out (KO) and ASC KO BMDMs were infected with RSV (1 MOI) for 12 h. The cell lysate from mock infected and RSV infected cells were subjected to Western blot analysis with mouse caspase-1 p10 subunit specific antibody. ( B ) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in RSV infected WT, NLRP3 KO and ASC KO BMDMs. The gels shown in (A) and (B) is representative of three independent experiments that yielded similar results.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: NLRP3 and ASC are required for caspase-1 activation following RSV infection. ( A ) Wild type (WT), NLRP3 knock-out (KO) and ASC KO BMDMs were infected with RSV (1 MOI) for 12 h. The cell lysate from mock infected and RSV infected cells were subjected to Western blot analysis with mouse caspase-1 p10 subunit specific antibody. ( B ) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in RSV infected WT, NLRP3 KO and ASC KO BMDMs. The gels shown in (A) and (B) is representative of three independent experiments that yielded similar results.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Activation Assay, Infection, Knock-Out, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing

    Caspase-1 dependent IL-1β release during RSV infection. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β and/or pro-caspase-1 plasmids. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Human macrophagic U937 cells were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( C ) Primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( D ) Wild type (WT) or caspase-1 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: Caspase-1 dependent IL-1β release during RSV infection. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β and/or pro-caspase-1 plasmids. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Human macrophagic U937 cells were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( C ) Primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( D ) Wild type (WT) or caspase-1 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Transfection, Enzyme-linked Immunosorbent Assay, Derivative Assay, Knock-Out, Standard Deviation

    A schematic model depicting the mechanism of IL-1β secretion during RSV infection. RSV infection activates TLR2/MyD88 pathway that culminates in NF-κB activation. Nuclear translocation of NF-κB results in NF-κB mediated trans-activation of pro-IL-1β and NLRP3 genes. Intracellular ROS generated during RSV infection and potassium efflux due to stimulation of ATP-sensitive ion channel triggers assembly of NLRP3/ASC inflammasome complex. NLRP3/ASC inflammasome activates caspase-1, which subsequently cleaves pro-IL-1β protein into its mature form. Mature IL-1β is secreted from the cells.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: A schematic model depicting the mechanism of IL-1β secretion during RSV infection. RSV infection activates TLR2/MyD88 pathway that culminates in NF-κB activation. Nuclear translocation of NF-κB results in NF-κB mediated trans-activation of pro-IL-1β and NLRP3 genes. Intracellular ROS generated during RSV infection and potassium efflux due to stimulation of ATP-sensitive ion channel triggers assembly of NLRP3/ASC inflammasome complex. NLRP3/ASC inflammasome activates caspase-1, which subsequently cleaves pro-IL-1β protein into its mature form. Mature IL-1β is secreted from the cells.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Activation Assay, Translocation Assay, Generated

    ASC expression is essential for IL-1β production during RSV infection. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and ASC plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Wild type (WT) or ASC knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: ASC expression is essential for IL-1β production during RSV infection. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and ASC plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Wild type (WT) or ASC knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Expressing, Infection, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Knock-Out, Standard Deviation

    Potassium efflux plays a critical role in IL-1β production during RSV infection. ( A ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ATP-sensitive potassium channel inhibitor glibenclamide (50 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Cells were also treated with LPS and ATP in the presence of either DMSO or glibenclamide. ( B ) U937 cells were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( C ) BMDM were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 6 h and 12 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: Potassium efflux plays a critical role in IL-1β production during RSV infection. ( A ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ATP-sensitive potassium channel inhibitor glibenclamide (50 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Cells were also treated with LPS and ATP in the presence of either DMSO or glibenclamide. ( B ) U937 cells were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. ( C ) BMDM were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 6 h and 12 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    TLR2/MyD88 pathway is required for IL-1β release during RSV infection. ( A ) Wild type (WT), TLR2 knock-out (KO) and TLR4 KO BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) Wild type (WT) and MyD88 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( C ) RT-PCR analysis of pro-IL-1β expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. (D) RT-PCR analysis of caspase-1 and NLRP3 expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. The gels shown in (C) and (D) are representative of three independent experiments that yielded similar results. For ELISA assay, each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: TLR2/MyD88 pathway is required for IL-1β release during RSV infection. ( A ) Wild type (WT), TLR2 knock-out (KO) and TLR4 KO BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) Wild type (WT) and MyD88 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( C ) RT-PCR analysis of pro-IL-1β expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. (D) RT-PCR analysis of caspase-1 and NLRP3 expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. The gels shown in (C) and (D) are representative of three independent experiments that yielded similar results. For ELISA assay, each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Knock-Out, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Standard Deviation

    ROS produced during RSV infection triggers IL-1β secretion. ( A ) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor APDC (50 µM). Water served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (10 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( C ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (2 µM) or DMSO (vehicle control). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: ROS produced during RSV infection triggers IL-1β secretion. ( A ) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor APDC (50 µM). Water served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (10 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( C ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (2 µM) or DMSO (vehicle control). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Produced, Infection, Enzyme-linked Immunosorbent Assay, Derivative Assay, Standard Deviation

    NF-κB signaling is critical for IL-1β secretion during RSV infection. ( A ) Human U937 cells were infected with RSV (1 MOI) in the presence of either DMSO (control) or NF-κB inhibitor (BAY 11-7082). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) RT-PCR analysis of pro-IL-1β, caspase-1 and NLRP3 expression in U937 cells infected with RSV in the presence of either DMSO (control) or BAY 11-7082. ( C ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either DMSO (control) or BAY 11-7082. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( D ) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in WT BMDMs infected with RSV in the presence of either DMSO (control) or BAY 11-7082. The gels shown in (B) and (D) are representative of three independent experiments that yielded similar results. For ELISA results, each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: NF-κB signaling is critical for IL-1β secretion during RSV infection. ( A ) Human U937 cells were infected with RSV (1 MOI) in the presence of either DMSO (control) or NF-κB inhibitor (BAY 11-7082). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( B ) RT-PCR analysis of pro-IL-1β, caspase-1 and NLRP3 expression in U937 cells infected with RSV in the presence of either DMSO (control) or BAY 11-7082. ( C ) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either DMSO (control) or BAY 11-7082. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. ( D ) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in WT BMDMs infected with RSV in the presence of either DMSO (control) or BAY 11-7082. The gels shown in (B) and (D) are representative of three independent experiments that yielded similar results. For ELISA results, each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Derivative Assay, Standard Deviation

    IL-1β secretion during RSV infection requires NLRP3 expression. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and NLRP3 plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Wild type (WT) or NLRP3 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Journal: PLoS ONE

    Article Title: TLR2/MyD88/NF-?B Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

    doi: 10.1371/journal.pone.0029695

    Figure Lengend Snippet: IL-1β secretion during RSV infection requires NLRP3 expression. ( A ) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and NLRP3 plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. ( B ) Wild type (WT) or NLRP3 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.

    Article Snippet: Interestingly, we failed to detect IL-1β secretion from RSV infected human epithelial cell-lines [human lung epithelial A549 (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells and Hela (obtained from American Type Culture Collection (ATCC), Manassas, VA) cells (data not shown).

    Techniques: Infection, Expressing, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Knock-Out, Standard Deviation

    Co-infection elicited reduction in the number of CD11b or CD11c myeloid cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. to determine the frequency of NK1.1 gated on CD3 negative cells. (A) Proportion of CD11b myeloid cells, which was significantly low in the I + R group on day 2 was increased by day 7. (B) Proportions of CD11c myeloid cells were lower in I + R group compared to other groups on days 2 and 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited reduction in the number of CD11b or CD11c myeloid cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. to determine the frequency of NK1.1 gated on CD3 negative cells. (A) Proportion of CD11b myeloid cells, which was significantly low in the I + R group on day 2 was increased by day 7. (B) Proportions of CD11c myeloid cells were lower in I + R group compared to other groups on days 2 and 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited very low numbers of CD8 and CD8+CD25 frequencies during the course of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD8 T cells proportions were very low in I + R group compared to other groups. (B) The frequency of CD8+CD25 T cells was very low in I + R group compared to other groups. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited very low numbers of CD8 and CD8+CD25 frequencies during the course of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD8 T cells proportions were very low in I + R group compared to other groups. (B) The frequency of CD8+CD25 T cells was very low in I + R group compared to other groups. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited high proportions of NKT cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) Proportions of NK1.1 cells gated on CD3 on day 2. (B) Frequency of NKT cells on day 4. (C) Frequency of NKT cells on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited high proportions of NKT cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) Proportions of NK1.1 cells gated on CD3 on day 2. (B) Frequency of NKT cells on day 4. (C) Frequency of NKT cells on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited very high numbers of CD4 and CD4+CD25 frequencies towards the later stages of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD4 T cells proportions were high on days 2 and 7 in I + R group. (B) The frequency of CD4+CD25 T cells was highest in I + R on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited very high numbers of CD4 and CD4+CD25 frequencies towards the later stages of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD4 T cells proportions were high on days 2 and 7 in I + R group. (B) The frequency of CD4+CD25 T cells was highest in I + R on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Influenza A virus (IAV) M, PB1 and PB2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). (A) On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of IAV genes. (B) mRNA expression level of M gene (C) mRNA expression level of PB1 (D) mRNA expression level of PB2. IAV M, PB1 and PB2 genes were expressed early on infection in I + R group and this was sustained up until the 4 th day. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for IAV M, PB1 and PB2 were not detected in RSV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Influenza A virus (IAV) M, PB1 and PB2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). (A) On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of IAV genes. (B) mRNA expression level of M gene (C) mRNA expression level of PB1 (D) mRNA expression level of PB2. IAV M, PB1 and PB2 genes were expressed early on infection in I + R group and this was sustained up until the 4 th day. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for IAV M, PB1 and PB2 were not detected in RSV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Infection

    Respiratory syncytial virus (RSV) NS1 and NS2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of NS1 gene (B) mRNA expression level of NS2. RSV NS1 and NS2 expressions were downregulated by day 7 in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV NS1 and NS2 were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Respiratory syncytial virus (RSV) NS1 and NS2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of NS1 gene (B) mRNA expression level of NS2. RSV NS1 and NS2 expressions were downregulated by day 7 in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV NS1 and NS2 were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Infection

    Respiratory syncytial virus (RSV) F, G and M genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of F gene (B) mRNA expression level of M (C) mRNA expression level of G. RSV F and G expression were markedly downregulated in I + R group by day 7 while RSV M expression was very low throughout the infection period in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV F, M and G were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Respiratory syncytial virus (RSV) F, G and M genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of F gene (B) mRNA expression level of M (C) mRNA expression level of G. RSV F and G expression were markedly downregulated in I + R group by day 7 while RSV M expression was very low throughout the infection period in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV F, M and G were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Infection

    Co-infection elicited reduction in the number of CD11b or CD11c myeloid cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. to determine the frequency of NK1.1 gated on CD3 negative cells. (A) Proportion of CD11b myeloid cells, which was significantly low in the I + R group on day 2 was increased by day 7. (B) Proportions of CD11c myeloid cells were lower in I + R group compared to other groups on days 2 and 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited reduction in the number of CD11b or CD11c myeloid cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. to determine the frequency of NK1.1 gated on CD3 negative cells. (A) Proportion of CD11b myeloid cells, which was significantly low in the I + R group on day 2 was increased by day 7. (B) Proportions of CD11c myeloid cells were lower in I + R group compared to other groups on days 2 and 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited very low numbers of CD8 and CD8+CD25 frequencies during the course of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD8 T cells proportions were very low in I + R group compared to other groups. (B) The frequency of CD8+CD25 T cells was very low in I + R group compared to other groups. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited very low numbers of CD8 and CD8+CD25 frequencies during the course of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD8 T cells proportions were very low in I + R group compared to other groups. (B) The frequency of CD8+CD25 T cells was very low in I + R group compared to other groups. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited high proportions of NKT cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) Proportions of NK1.1 cells gated on CD3 on day 2. (B) Frequency of NKT cells on day 4. (C) Frequency of NKT cells on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited high proportions of NKT cells. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) Proportions of NK1.1 cells gated on CD3 on day 2. (B) Frequency of NKT cells on day 4. (C) Frequency of NKT cells on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Co-infection elicited very high numbers of CD4 and CD4+CD25 frequencies towards the later stages of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD4 T cells proportions were high on days 2 and 7 in I + R group. (B) The frequency of CD4+CD25 T cells was highest in I + R on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Co-infection elicited very high numbers of CD4 and CD4+CD25 frequencies towards the later stages of infection. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7 splenic cells (n = 5) were processed for flow cytometric analysis. (A) CD4 T cells proportions were high on days 2 and 7 in I + R group. (B) The frequency of CD4+CD25 T cells was highest in I + R on day 7. Unless indicated by lines, a vs uninfected, b vs IAV (I), c vs RSV (R), d vs IR, e vs RI. No data was available for IR and RI as well as for IAV (I), RSV(R) and I + R groups on day 2 and day 4 respectively.

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Infection, Mouse Assay, Flow Cytometry

    Respiratory syncytial virus (RSV) NS1 and NS2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of NS1 gene (B) mRNA expression level of NS2. RSV NS1 and NS2 expressions were downregulated by day 7 in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV NS1 and NS2 were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Respiratory syncytial virus (RSV) NS1 and NS2 genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of NS1 gene (B) mRNA expression level of NS2. RSV NS1 and NS2 expressions were downregulated by day 7 in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV NS1 and NS2 were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Infection

    Respiratory syncytial virus (RSV) F, G and M genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of F gene (B) mRNA expression level of M (C) mRNA expression level of G. RSV F and G expression were markedly downregulated in I + R group by day 7 while RSV M expression was very low throughout the infection period in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV F, M and G were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Journal: Heliyon

    Article Title: Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice

    doi: 10.1016/j.heliyon.2018.e01094

    Figure Lengend Snippet: Respiratory syncytial virus (RSV) F, G and M genes expressions across the experimental groups. Six groups of BALB/c mice were intranasally treated with 32 μl of normal saline (uninfected), RSV A2 (RSV(R)), Pr/H3N2 (IAV (I)) or co-treated simultaneously (I + R) or one after the other at 24 hours interval (IR and RI). On days 2, 4 and 7, mRNA was extracted from lungs (n = 5) for real time PCR analysis of RSV genes. (A) mRNA expression level of F gene (B) mRNA expression level of M (C) mRNA expression level of G. RSV F and G expression were markedly downregulated in I + R group by day 7 while RSV M expression was very low throughout the infection period in I + R group. CT values were normalized with GADPH, mRNA expression level was calculated using 2 −ΔΔCT method and presented as Log10 fold increase relative to uninfected controls. mRNA expressions for RSV F, M and G were not detected in IAV (single virus) infected mice. * vs Day 2 and # vs Day 4 for within group comparison while a vs RI, b vs IR and c vs I + R for comparing between groups of the same day (p

    Article Snippet: Human RSV A2 (ATCC-1540) was obtained from the National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire, EN6 3QG, WHO International Laboratory for Biological Standard, UK Official Medicine Control Laboratory, and was propagated on Hep-2 cell line.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Infection

    RSV filaments are formed by a rapid vesicular extension and retraction motion. a Vero cells were infected with RSV A2 at an MOI of 1. At 16 hpi, SBA-488 was delivered and allowed to internalize for an hour. Cells were then imaged live at 12.13 Hz. Time series are shown with either even temporal spacing ( top ) or as selected frames from the movie ( bottom ). Yellow arrowheads indicate distension of vesicles, while arrows indicate extended filaments and branched filaments. Scale bar represents 5 µm. b Individual extension and retraction events were analyzed by hand on a frame-by-frame basis to determine the mean velocity. 30 events of each type were analyzed. Error bars represent standard deviation. c Vero cells were infected with RSV A2 at an MOI of 1. After 12 h, SBA-488 ( green ) was delivered to the cells and allowed to internalize for 1 h before being imaged every 10 min. Cropped regions of two individual filaments are shown. Yellow arrows mark the two apparent ends of the filaments. Scale bars represent 1 µm

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: RSV filaments are formed by a rapid vesicular extension and retraction motion. a Vero cells were infected with RSV A2 at an MOI of 1. At 16 hpi, SBA-488 was delivered and allowed to internalize for an hour. Cells were then imaged live at 12.13 Hz. Time series are shown with either even temporal spacing ( top ) or as selected frames from the movie ( bottom ). Yellow arrowheads indicate distension of vesicles, while arrows indicate extended filaments and branched filaments. Scale bar represents 5 µm. b Individual extension and retraction events were analyzed by hand on a frame-by-frame basis to determine the mean velocity. 30 events of each type were analyzed. Error bars represent standard deviation. c Vero cells were infected with RSV A2 at an MOI of 1. After 12 h, SBA-488 ( green ) was delivered to the cells and allowed to internalize for 1 h before being imaged every 10 min. Cropped regions of two individual filaments are shown. Yellow arrows mark the two apparent ends of the filaments. Scale bars represent 1 µm

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Infection, Standard Deviation

    Vesicular RSV filament formation is microtubule and dynein dependent. a Vero cells were infected with RSV A2 at an MOI of 1. At 12 hpi, SBA-488 was delivered to the cells and allowed to internalize for 1 h. Cells were then imaged live briefly at 9 Hz prior to drug delivery to verify the presence vesicular filament formation. Cells were then treated with nocodazole to disrupt microtubules, cytochalasin D to disrupt actin filaments, tetracaine to inhibit molecular motors, or EHNA to inhibit dynein before being imaged live again. Data is presented as SD maps over 30 s of imaging. Yellow arrows indicate fast motion, shown as streaks, while yellow arrowheads indicate slow motion, shown as hollow circles . Scale bars represent 5 µm. b Tubulin-GFP ( green ) was expressed in Vero cells overnight prior to infection with RSV A2 at an MOI of 1. At 12 hpi, SBA-647 ( red ) was delivered and allowed to internalize for 1 h. Cells were then imaged live at 4.9 Hz. Scale bar represents 3 µm

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: Vesicular RSV filament formation is microtubule and dynein dependent. a Vero cells were infected with RSV A2 at an MOI of 1. At 12 hpi, SBA-488 was delivered to the cells and allowed to internalize for 1 h. Cells were then imaged live briefly at 9 Hz prior to drug delivery to verify the presence vesicular filament formation. Cells were then treated with nocodazole to disrupt microtubules, cytochalasin D to disrupt actin filaments, tetracaine to inhibit molecular motors, or EHNA to inhibit dynein before being imaged live again. Data is presented as SD maps over 30 s of imaging. Yellow arrows indicate fast motion, shown as streaks, while yellow arrowheads indicate slow motion, shown as hollow circles . Scale bars represent 5 µm. b Tubulin-GFP ( green ) was expressed in Vero cells overnight prior to infection with RSV A2 at an MOI of 1. At 12 hpi, SBA-647 ( red ) was delivered and allowed to internalize for 1 h. Cells were then imaged live at 4.9 Hz. Scale bar represents 3 µm

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Infection, Imaging

    SBA selectively binds RSV G. a Vero cells infected ( top ) or mock-infected ( bottom ) for 24 h with RSV A2 at an MOI of 1 were labeled with SBA-488 at 4 °C ( green ) before being fixed and stained for RSV G ( red ), RSV F ( blue ), and nuclei ( gray ). Extended focus image is shown. Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow . Scale bar represents 1 µm. c Cells transfected for 24 h with RSV G ( top ) and RSV F ( bottom ) mRNA were labeled with SBA-488 at 4 °C ( green ) before being fixed and stained without permeabilization for RSV G or RSV F ( red ) and nuclei ( gray ). Extended focus image is shown. Scale bar represents 15 µm. d Mander’s colocalization coefficient and Pearson’s correlation between RSV G or RSV F and SBA. Dotted line indicates 0 correlation. n = 16 cells per group. Error bars represent 95% confidence interval. Asterisks indicates p

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: SBA selectively binds RSV G. a Vero cells infected ( top ) or mock-infected ( bottom ) for 24 h with RSV A2 at an MOI of 1 were labeled with SBA-488 at 4 °C ( green ) before being fixed and stained for RSV G ( red ), RSV F ( blue ), and nuclei ( gray ). Extended focus image is shown. Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow . Scale bar represents 1 µm. c Cells transfected for 24 h with RSV G ( top ) and RSV F ( bottom ) mRNA were labeled with SBA-488 at 4 °C ( green ) before being fixed and stained without permeabilization for RSV G or RSV F ( red ) and nuclei ( gray ). Extended focus image is shown. Scale bar represents 15 µm. d Mander’s colocalization coefficient and Pearson’s correlation between RSV G or RSV F and SBA. Dotted line indicates 0 correlation. n = 16 cells per group. Error bars represent 95% confidence interval. Asterisks indicates p

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Infection, Labeling, Staining, Transfection

    SBA binding does not inhibit RSV binding or replication. a SBA-488 ( green ) was incubated with RSV A2 stocks for 30 min before filaments were isolated onto glass coverslips. Isolated virions were fixed and stained for RSV N ( red ) and RSV F ( blue ). Scale bar represents 2 µm. b SBA-488 labeled virus ( green ) was used to inoculate Vero cells at an MOI of 1. At 24 hpi, cells were fixed and immunostained for RSV N ( red ), RSV F ( blue ), and nuclei ( gray ). Scale bar represents 10 µm. c RSV A2 was incubated with either SBA or vehicle control for 2 h at 37 °C. Virus was then used to inoculate Vero cells at an MOI of 1. At 12 or 24 hpi, cells were lysed, and lysates were assayed by western blotting for RSV G. β-actin is included as a loading control. Image is representative of two independent experiments. d Replicate lanes ( n = 2) were quantified by densitometry to compare the amount of RSV G signal from SBA-labeled or unlabeled RSV infections. There were no statistically significant differences at either time point (two-way ANOVA, p > 0.05). Error bars represent standard deviation. e rgRSV-GFP was incubated with the indicated concentrations of either palivizumab, SBA, or vehicle control for 2 h at 37 °C. Virus treatments and a mock treatment were then used to inoculate Vero cells at an MOI of 1. At 48 hpi cells were lifted into a single-cell suspension and analyzed by flow cytometry. Data represents the mean percentage of infected cells in three independent experiments. Error bars represent standard deviation. Asterisk indicates significant difference vs. mock infection (one-way ANOVA on ranks, p

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: SBA binding does not inhibit RSV binding or replication. a SBA-488 ( green ) was incubated with RSV A2 stocks for 30 min before filaments were isolated onto glass coverslips. Isolated virions were fixed and stained for RSV N ( red ) and RSV F ( blue ). Scale bar represents 2 µm. b SBA-488 labeled virus ( green ) was used to inoculate Vero cells at an MOI of 1. At 24 hpi, cells were fixed and immunostained for RSV N ( red ), RSV F ( blue ), and nuclei ( gray ). Scale bar represents 10 µm. c RSV A2 was incubated with either SBA or vehicle control for 2 h at 37 °C. Virus was then used to inoculate Vero cells at an MOI of 1. At 12 or 24 hpi, cells were lysed, and lysates were assayed by western blotting for RSV G. β-actin is included as a loading control. Image is representative of two independent experiments. d Replicate lanes ( n = 2) were quantified by densitometry to compare the amount of RSV G signal from SBA-labeled or unlabeled RSV infections. There were no statistically significant differences at either time point (two-way ANOVA, p > 0.05). Error bars represent standard deviation. e rgRSV-GFP was incubated with the indicated concentrations of either palivizumab, SBA, or vehicle control for 2 h at 37 °C. Virus treatments and a mock treatment were then used to inoculate Vero cells at an MOI of 1. At 48 hpi cells were lifted into a single-cell suspension and analyzed by flow cytometry. Data represents the mean percentage of infected cells in three independent experiments. Error bars represent standard deviation. Asterisk indicates significant difference vs. mock infection (one-way ANOVA on ranks, p

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Binding Assay, Incubation, Isolation, Staining, Labeling, Western Blot, Standard Deviation, Flow Cytometry, Cytometry, Infection

    RSV G recycles from the plasma membrane into intracellular vesicles. a Vero cells were infected with RSV A2 at an MOI of 1. Cells were either labeled with SBA-488 ( green ) at 12, 18, and 24 hpi ( top ) or with SBA-647 ( red ) at 12 hpi and SBA-488 at 24 hpi. Cells were then fixed and stained for RSV F ( blue ). Enlargements of cropped images of either filaments or intracellular vesicles from the white boxed regions with intensity profiles showing colocalization along the direction of the white arrow . Scale bar represents 10 µm. b Vero cells were infected with RSV for 12 hpi before being labeled with SBA-488 ( green ). Immediately following SBA delivery, cells were imaged live over 1 h. Extended focus images shown. Scale bars represent 10 µm

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: RSV G recycles from the plasma membrane into intracellular vesicles. a Vero cells were infected with RSV A2 at an MOI of 1. Cells were either labeled with SBA-488 ( green ) at 12, 18, and 24 hpi ( top ) or with SBA-647 ( red ) at 12 hpi and SBA-488 at 24 hpi. Cells were then fixed and stained for RSV F ( blue ). Enlargements of cropped images of either filaments or intracellular vesicles from the white boxed regions with intensity profiles showing colocalization along the direction of the white arrow . Scale bar represents 10 µm. b Vero cells were infected with RSV for 12 hpi before being labeled with SBA-488 ( green ). Immediately following SBA delivery, cells were imaged live over 1 h. Extended focus images shown. Scale bars represent 10 µm

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Infection, Labeling, Staining

    SBA allows for the visualization of RSV G dynamics in RSV infected cells. a Vero cells were infected with RSV A2 at an MOI of 1. At 12 hpi, cells were stained live with SBA-488 ( top ) or vehicle control ( bottom ), washed, and further incubated in complete growth media. 24 hpi, cells were fixed and immunostained for RSV N ( blue ), RSV F ( red ), and nuclei ( gray ). Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow . c Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 ( top, green ) or vehicle control ( bottom ), washed, and further incubated in complete growth media. 24 hpi, virus was harvested. Filaments were isolated onto glass and stained for RSV F ( blue ) and RSV G ( red ) Scale bar represents 2 µm. d Mean sum of RSV G intensity per filament was quantified. 50 isolated virions were analyzed for each condition. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05). e Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 or vehicle control, washed, and further incubated in complete growth media. 24 hpi, supernatants from single infected wells were titered by plaque assay in quadruplicate. Three wells were independently infected, stained, and assayed per treatment. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05)

    Journal: Nature Communications

    Article Title: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

    doi: 10.1038/s41467-017-00732-z

    Figure Lengend Snippet: SBA allows for the visualization of RSV G dynamics in RSV infected cells. a Vero cells were infected with RSV A2 at an MOI of 1. At 12 hpi, cells were stained live with SBA-488 ( top ) or vehicle control ( bottom ), washed, and further incubated in complete growth media. 24 hpi, cells were fixed and immunostained for RSV N ( blue ), RSV F ( red ), and nuclei ( gray ). Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow . c Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 ( top, green ) or vehicle control ( bottom ), washed, and further incubated in complete growth media. 24 hpi, virus was harvested. Filaments were isolated onto glass and stained for RSV F ( blue ) and RSV G ( red ) Scale bar represents 2 µm. d Mean sum of RSV G intensity per filament was quantified. 50 isolated virions were analyzed for each condition. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05). e Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 or vehicle control, washed, and further incubated in complete growth media. 24 hpi, supernatants from single infected wells were titered by plaque assay in quadruplicate. Three wells were independently infected, stained, and assayed per treatment. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05)

    Article Snippet: Human RSV A2 (ATCC VR-1544) or rgRSV-GFP (a gift from Martin L. Moore’s lab, Emory University) was propagated in HEp-2 cells when the cells were > 80% confluent.

    Techniques: Infection, Staining, Incubation, Isolation, Standard Deviation, Labeling, MANN-WHITNEY, Plaque Assay