recombinant influenza ha proteins  (Sino Biological)


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    Structured Review

    Sino Biological recombinant influenza ha proteins
    Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against <t>recombinant</t> <t>HA</t> from H5, H6, H8, H9, H11, H13, and H1 (PR8) <t>influenza</t> viruses (mean ± SEM). ** p
    Recombinant Influenza Ha Proteins, 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
    https://www.bioz.com/result/recombinant influenza ha proteins/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant influenza ha proteins - by Bioz Stars, 2021-07
    94/100 stars

    Images

    1) Product Images from "Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza"

    Article Title: Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1701088

    Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against recombinant HA from H5, H6, H8, H9, H11, H13, and H1 (PR8) influenza viruses (mean ± SEM). ** p
    Figure Legend Snippet: Increased strain-specific IgG titers after a single MHCII-targeted vaccination. BALB/c mice ( n = 6 per group) were immunized once with 25 μg of plasmids encoding one of the indicated vaccines. ( A ) Sera were harvested at weeks 2, 3, and 4 after vaccination, and IgG responses in sera from each of the vaccine groups were measured by ELISAs against recombinant HA from H5, H6, H8, H9, H11, H13, and H1 (PR8) influenza viruses (mean ± SEM). ** p

    Techniques Used: Mouse Assay, Recombinant

    Ab responses against all included HAs after DNA immunization with vaccine mixtures. BALB/c mice ( n = 6 per group) were immunized three times (weeks 0, 4, and 12) with 25 μg of DNA of the indicated vaccine (arrowheads). Serum samples were harvested 2–3 wk after each vaccine delivery, and HA-specific IgG responses were measured in sandwich ELISAs against recombinant HA from H5, H6, H8, H9, H11, and H13 influenza viruses (mean ± SEM). * p
    Figure Legend Snippet: Ab responses against all included HAs after DNA immunization with vaccine mixtures. BALB/c mice ( n = 6 per group) were immunized three times (weeks 0, 4, and 12) with 25 μg of DNA of the indicated vaccine (arrowheads). Serum samples were harvested 2–3 wk after each vaccine delivery, and HA-specific IgG responses were measured in sandwich ELISAs against recombinant HA from H5, H6, H8, H9, H11, and H13 influenza viruses (mean ± SEM). * p

    Techniques Used: Mouse Assay, Recombinant

    2) Product Images from "Pan-Influenza A Protection by Prime–Boost Vaccination with Cold-Adapted Live-Attenuated Influenza Vaccine in a Mouse Model"

    Article Title: Pan-Influenza A Protection by Prime–Boost Vaccination with Cold-Adapted Live-Attenuated Influenza Vaccine in a Mouse Model

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00116

    Cross-reactive hemagglutinin (HA)-specific antibody responses elicited by vaccination. (A) Breadth of the HA-specific sera IgG antibodies. To examine the breadth of HA-specific sera IgG antibodies induced by vaccination, recombinant HA proteins expressed in insect cells were used as coating antigens in ELISA. The HA proteins tested include five different group 1 HAs from H1N1 (A/California/6/2009), H1N1 (A/Puerto Rico/8/1934), H2N2 (A/Canada/720/2006), H5N1 (A/Indonesia/5/2005), and H9N2 (A/Hong Kong/35820/2009) and two group 2 HAs from H3N2 (A/Sydney/5/1997) and H7N9 (A/Anhui/1/2013) influenza viruses. (B) Antibody titers specific to HA full-length or stalk of pH1N1 or PR8 (H1N1) virus. Using the Escherichia coli -expressed LysRS-HA fusion proteins as coating antigens, sera IgG antibodies specific to the HA full-length or stalk protein were measured by ELISA. Antibody titers were expressed as the reciprocal serum dilution that yielded OD 450 greater than the mean + 2 SD (SD) of PBS control group. Data are the mean of each cohort ( N = 5), and error bars indicate SD. *** P
    Figure Legend Snippet: Cross-reactive hemagglutinin (HA)-specific antibody responses elicited by vaccination. (A) Breadth of the HA-specific sera IgG antibodies. To examine the breadth of HA-specific sera IgG antibodies induced by vaccination, recombinant HA proteins expressed in insect cells were used as coating antigens in ELISA. The HA proteins tested include five different group 1 HAs from H1N1 (A/California/6/2009), H1N1 (A/Puerto Rico/8/1934), H2N2 (A/Canada/720/2006), H5N1 (A/Indonesia/5/2005), and H9N2 (A/Hong Kong/35820/2009) and two group 2 HAs from H3N2 (A/Sydney/5/1997) and H7N9 (A/Anhui/1/2013) influenza viruses. (B) Antibody titers specific to HA full-length or stalk of pH1N1 or PR8 (H1N1) virus. Using the Escherichia coli -expressed LysRS-HA fusion proteins as coating antigens, sera IgG antibodies specific to the HA full-length or stalk protein were measured by ELISA. Antibody titers were expressed as the reciprocal serum dilution that yielded OD 450 greater than the mean + 2 SD (SD) of PBS control group. Data are the mean of each cohort ( N = 5), and error bars indicate SD. *** P

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay

    Experimental design of vaccination and challenge in mice. (A) Vaccination schedule and design of prime–boost vaccination against heterologous challenge. Four different prime–boost vaccination groups were designed using three different cold-adapted, live attenuated influenza vaccines (CAIVs), ca-pH1N1, ca-NCH1N1, and ca-IDH5N1. Prime and boost CAIVs (10 5 PFUs of each vaccine) were administered into mice via intranasal route with two weeks interval. A month later, each group was divided into four subgroups (20 subgroups in total) and challenged with 10 mouse lethal dose 50 (MLD 50 ) of each of four heterologous influenza viruses. (B) Phylogenetic tree of influenza A hemagglutinin (HA) proteins. HA to which binding affinity of vaccination-induced antibodies or protection efficacy in vivo tested in this study was highlighted in colored circles.
    Figure Legend Snippet: Experimental design of vaccination and challenge in mice. (A) Vaccination schedule and design of prime–boost vaccination against heterologous challenge. Four different prime–boost vaccination groups were designed using three different cold-adapted, live attenuated influenza vaccines (CAIVs), ca-pH1N1, ca-NCH1N1, and ca-IDH5N1. Prime and boost CAIVs (10 5 PFUs of each vaccine) were administered into mice via intranasal route with two weeks interval. A month later, each group was divided into four subgroups (20 subgroups in total) and challenged with 10 mouse lethal dose 50 (MLD 50 ) of each of four heterologous influenza viruses. (B) Phylogenetic tree of influenza A hemagglutinin (HA) proteins. HA to which binding affinity of vaccination-induced antibodies or protection efficacy in vivo tested in this study was highlighted in colored circles.

    Techniques Used: Mouse Assay, Binding Assay, In Vivo

    Related Articles

    Recombinant:

    Article Title: Induction of Cross-Reactive and Protective Antibody Responses After DNA Vaccination With MHCII-Targeted Stem Domain From Influenza Hemagglutinin
    Article Snippet: ELISA for Detection of Serum Antibodies Blood was harvested by puncture of the saphenous vein, and sera were collected by centrifugation. .. Ninety-six-well plates (Costar 3590) were coated with inactivated A/PR/8/34 (H1N1) (PR8) virus (1:1600) (virus supplied from Charles River, USA, with HA titer 1:65,536 in 0.05 ml) in PBS azide, or with recombinant HA protein (1 μg/ml) from one of the following viruses: PR8 (11684-V08H, Sino Biological Inc., USA), A/Hong Kong/1/1968 (H3N2) (40116-V08H1, Sino Biological Inc.), A/Hong Kong/483/97 (H5N1) (11689-V08H, Sino Biological Inc.), A/Shanghai/1/2013 (H7N9) (40104-V08H, Sino Biological Inc.), or A/Hong Kong/1073/99 (H9N2) (11229-V08H, Sino Biological Inc.). .. Next, plates were incubated with either alkaline phosphatase conjugated anti-mouse IgG (A1418, Sigma-Aldrich, Germany), biotinylated anti-IgG1a (553599, BD Pharmingen), or biotinylated anti-IgG2aa (553502, BD Pharmingen), and developed as above.

    Article Title: Targeting of HA to chemokine receptors induces strong and cross-reactive T cell responses after DNA vaccination in pigs.
    Article Snippet: Serum ELISASera were isolated from blood by centrifugation. .. ELISA 96-well plates (Costar 3590, Sigma-Aldrich) were coated with either recombinant HA from PR8 (11684-V08H, Sino Biological Inc., North Wales, PA, US), rec. .. HA from A/Anhui/1/2005 (H5N1) (11048- V08H1, Sino Biological Inc.), rec.

    Article Title: Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza
    Article Snippet: .. ELISA 96-well plates (Costar 3590) were coated with one of the following recombinant influenza HA proteins: PR8 (11684-V08H; Sino Biological), A/Hong Kong/483/97 (H5N1) (11689-V08H; Sino Biological), A/northern shoveler/California/HKWF115/07 (H6N1) (MBS434125; MyBioSource, San Diego, CA, or 11723-V08H; Sino Biological), A/pintail duck/Alberta/114/1979 (H8N4) (11722-V08H; Sino Biological), A/Hong Kong/1073/99 (H9N2) (11229-V08H; Sino Biological), A/duck/Yangzhou/906/2002 (H11N2) (11705-V08H; Sino Biological), A/black-headed gull/Netherlands/1/00 (H13N8) (11721-V08H; Sino Biological), A/Shanghai/1/2013 (H7N9) (40104-V08B; Sino Biological), or A/Hong Kong/1/1968 (H3N2) (40116-V08B; Sino Biological). .. Alternatively, plates were coated with inactivated virus: PR8 (1:1600; Charles River, Wilmington, MA) or Cal07 (Pandemrix; 1:50; GlaxoSmithKline, Brentford, U.K.).

    Article Title: Induction of Cross-Reactive and Protective Antibody Responses After DNA Vaccination With MHCII-Targeted Stem Domain From Influenza Hemagglutinin
    Article Snippet: .. The ELISAs were set up with coats of HA protein from influenza PR8 (11684-V08H, Sino Biological Inc., USA) or A/Hong Kong/483/97 (H5N1) (11689-V08H, Sino Biological Inc.), or ovalbumin (OVA) (A5503, Sigma), or by coating with Phox-BSA and recombinant proteins expressing Phox-specific scFv linked to the HA stem. .. Detection was performed with either alkaline phosphatase-conjugated anti-mouse IgG (A1418 Sigma-Aldrich) or biotinylated anti-IgM (553515, BD Pharmingen) and streptavidin alkaline phosphatase (1:3000, GE Healthcare).

    Article Title: Nucleoside-modified mRNA vaccines induce potent T follicular helper and germinal center B cell responses
    Article Snippet: The blocking protein for ELISA assays was BSA (Sigma). .. The recombinant PR8 HA protein was purchased from Sino Biological Inc. (catalog number 11684-V08H) and was reconstituted in sterile PBS. .. The MF59 adjuvant (AddaVax) was purchased from InvivoGen.

    Article Title: Toxicity and Immunogenicity of a Tardigrade Cytosolic Abundant Heat Soluble Protein in Mice
    Article Snippet: The first group was given Lactated Ringer’s solution as a negative control. .. The next two groups were given low (1.25 g/L) or high (15 g/L) doses of CAHS D. The final group was given 0.05 g/L recombinant hemagglutinin protein (HA) from the influenza A H1N1 (A/Puerto Rico/8/1934) flu virus (Sino Biological 11684-V08H). ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Targeting of HA to chemokine receptors induces strong and cross-reactive T cell responses after DNA vaccination in pigs.
    Article Snippet: Serum ELISASera were isolated from blood by centrifugation. .. ELISA 96-well plates (Costar 3590, Sigma-Aldrich) were coated with either recombinant HA from PR8 (11684-V08H, Sino Biological Inc., North Wales, PA, US), rec. .. HA from A/Anhui/1/2005 (H5N1) (11048- V08H1, Sino Biological Inc.), rec.

    Article Title: Simultaneous Targeting of Multiple Hemagglutinins to APCs for Induction of Broad Immunity against Influenza
    Article Snippet: .. ELISA 96-well plates (Costar 3590) were coated with one of the following recombinant influenza HA proteins: PR8 (11684-V08H; Sino Biological), A/Hong Kong/483/97 (H5N1) (11689-V08H; Sino Biological), A/northern shoveler/California/HKWF115/07 (H6N1) (MBS434125; MyBioSource, San Diego, CA, or 11723-V08H; Sino Biological), A/pintail duck/Alberta/114/1979 (H8N4) (11722-V08H; Sino Biological), A/Hong Kong/1073/99 (H9N2) (11229-V08H; Sino Biological), A/duck/Yangzhou/906/2002 (H11N2) (11705-V08H; Sino Biological), A/black-headed gull/Netherlands/1/00 (H13N8) (11721-V08H; Sino Biological), A/Shanghai/1/2013 (H7N9) (40104-V08B; Sino Biological), or A/Hong Kong/1/1968 (H3N2) (40116-V08B; Sino Biological). .. Alternatively, plates were coated with inactivated virus: PR8 (1:1600; Charles River, Wilmington, MA) or Cal07 (Pandemrix; 1:50; GlaxoSmithKline, Brentford, U.K.).

    Expressing:

    Article Title: Induction of Cross-Reactive and Protective Antibody Responses After DNA Vaccination With MHCII-Targeted Stem Domain From Influenza Hemagglutinin
    Article Snippet: .. The ELISAs were set up with coats of HA protein from influenza PR8 (11684-V08H, Sino Biological Inc., USA) or A/Hong Kong/483/97 (H5N1) (11689-V08H, Sino Biological Inc.), or ovalbumin (OVA) (A5503, Sigma), or by coating with Phox-BSA and recombinant proteins expressing Phox-specific scFv linked to the HA stem. .. Detection was performed with either alkaline phosphatase-conjugated anti-mouse IgG (A1418 Sigma-Aldrich) or biotinylated anti-IgM (553515, BD Pharmingen) and streptavidin alkaline phosphatase (1:3000, GE Healthcare).

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    Sino Biological rabbit anti ha polyclonal antibody
    M2 interacts with TRAPPC6A and TRAPPC6AΔ in mammalian cells. (A to C) Plasmids expressing TRAPPC6A-Myc and Flag-SC09M2 (A), TRAPPC6A-Myc and Flag-AH05M2 (B), or TRAPPC6A-Myc and Flag-WSNM2 (C) were transfected individually or in combination, as indicated, in HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Flag MAb or a mouse anti-Myc MAb and were subjected to Western blotting with a rabbit anti-Flag <t>polyclonal</t> antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (D) Western blotting of proteins bound to GST alone or to GST-TRAPPC6A. HEK293T cells transfected with pCAGGS-SC09M2 or with the pCAGGS vector were lysed with IP buffer, and the lysate was incubated with purified GST or GST-TRAPPC6A and then subjected to a pulldown assay. Equal volumes of proteins bound to the beads and the original cell lysates (5% of the input) were examined by Western blotting using a mouse anti-M2 MAb or a mouse anti-actin MAb, respectively. The GST-tagged proteins in the eluates were detected by Coomassie blue (CB) staining. (E) Confocal analysis of the distribution of M2 and TRAPPC6A proteins in A549 cells. pCAGGS-TRAPPC6A-myc and pCAGGS-Flag-SC09M2 were transfected individually or in combination into A549 cells and assessed by immunofluorescence staining. IAV M2 was detected with a mouse anti-Flag MAb and visualized with Alexa Fluor 488 (green). TRAPPC6A was detected with a rabbit anti-Myc polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of Alexa Fluor 546 and 488 in the merged image. (F) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1 or pEGFP-C1-BM2 into HEK293T cells for 48 h before the cells were lysed. Following immunoprecipitation of the cell lysates with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of BM2 and TRAPPC6A, respectively. (G) Co-IP of M2 and TRAPPC6AΔ. pCAGGS-Flag-SC09M2 was cotransfected with pCAGGS-TRAPPC6A-myc or pCAGGS-TRAPPC6AΔ-myc into HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Myc MAb and subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A or TRAPPC6AΔ, respectively.
    Rabbit Anti Ha Polyclonal Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti ha polyclonal antibody/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ha polyclonal antibody - by Bioz Stars, 2021-07
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    80
    Sino Biological recombinant mex4108 ha protein plasma
    Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following <t>MEX4108</t> infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p
    Recombinant Mex4108 Ha Protein Plasma, supplied by Sino Biological, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Sino Biological influenza a h1n1 hemagglutinin ha antibody mouse mab
    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, <t>influenza</t> A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.
    Influenza A H1n1 Hemagglutinin Ha Antibody Mouse Mab, 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
    https://www.bioz.com/result/influenza a h1n1 hemagglutinin ha antibody mouse mab/product/Sino Biological
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    Image Search Results


    M2 interacts with TRAPPC6A and TRAPPC6AΔ in mammalian cells. (A to C) Plasmids expressing TRAPPC6A-Myc and Flag-SC09M2 (A), TRAPPC6A-Myc and Flag-AH05M2 (B), or TRAPPC6A-Myc and Flag-WSNM2 (C) were transfected individually or in combination, as indicated, in HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Flag MAb or a mouse anti-Myc MAb and were subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (D) Western blotting of proteins bound to GST alone or to GST-TRAPPC6A. HEK293T cells transfected with pCAGGS-SC09M2 or with the pCAGGS vector were lysed with IP buffer, and the lysate was incubated with purified GST or GST-TRAPPC6A and then subjected to a pulldown assay. Equal volumes of proteins bound to the beads and the original cell lysates (5% of the input) were examined by Western blotting using a mouse anti-M2 MAb or a mouse anti-actin MAb, respectively. The GST-tagged proteins in the eluates were detected by Coomassie blue (CB) staining. (E) Confocal analysis of the distribution of M2 and TRAPPC6A proteins in A549 cells. pCAGGS-TRAPPC6A-myc and pCAGGS-Flag-SC09M2 were transfected individually or in combination into A549 cells and assessed by immunofluorescence staining. IAV M2 was detected with a mouse anti-Flag MAb and visualized with Alexa Fluor 488 (green). TRAPPC6A was detected with a rabbit anti-Myc polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of Alexa Fluor 546 and 488 in the merged image. (F) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1 or pEGFP-C1-BM2 into HEK293T cells for 48 h before the cells were lysed. Following immunoprecipitation of the cell lysates with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of BM2 and TRAPPC6A, respectively. (G) Co-IP of M2 and TRAPPC6AΔ. pCAGGS-Flag-SC09M2 was cotransfected with pCAGGS-TRAPPC6A-myc or pCAGGS-TRAPPC6AΔ-myc into HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Myc MAb and subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A or TRAPPC6AΔ, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: M2 interacts with TRAPPC6A and TRAPPC6AΔ in mammalian cells. (A to C) Plasmids expressing TRAPPC6A-Myc and Flag-SC09M2 (A), TRAPPC6A-Myc and Flag-AH05M2 (B), or TRAPPC6A-Myc and Flag-WSNM2 (C) were transfected individually or in combination, as indicated, in HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Flag MAb or a mouse anti-Myc MAb and were subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (D) Western blotting of proteins bound to GST alone or to GST-TRAPPC6A. HEK293T cells transfected with pCAGGS-SC09M2 or with the pCAGGS vector were lysed with IP buffer, and the lysate was incubated with purified GST or GST-TRAPPC6A and then subjected to a pulldown assay. Equal volumes of proteins bound to the beads and the original cell lysates (5% of the input) were examined by Western blotting using a mouse anti-M2 MAb or a mouse anti-actin MAb, respectively. The GST-tagged proteins in the eluates were detected by Coomassie blue (CB) staining. (E) Confocal analysis of the distribution of M2 and TRAPPC6A proteins in A549 cells. pCAGGS-TRAPPC6A-myc and pCAGGS-Flag-SC09M2 were transfected individually or in combination into A549 cells and assessed by immunofluorescence staining. IAV M2 was detected with a mouse anti-Flag MAb and visualized with Alexa Fluor 488 (green). TRAPPC6A was detected with a rabbit anti-Myc polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of Alexa Fluor 546 and 488 in the merged image. (F) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1 or pEGFP-C1-BM2 into HEK293T cells for 48 h before the cells were lysed. Following immunoprecipitation of the cell lysates with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of BM2 and TRAPPC6A, respectively. (G) Co-IP of M2 and TRAPPC6AΔ. pCAGGS-Flag-SC09M2 was cotransfected with pCAGGS-TRAPPC6A-myc or pCAGGS-TRAPPC6AΔ-myc into HEK293T cells. Forty-eight hours after transfection, cell lysates were immunoprecipitated with a mouse anti-Myc MAb and subjected to Western blotting with a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A or TRAPPC6AΔ, respectively.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Expressing, Transfection, Immunoprecipitation, Western Blot, Plasmid Preparation, Incubation, Purification, Staining, Immunofluorescence, Co-Immunoprecipitation Assay

    A leucine residue at position 96 of M2 is required for the TRAPPC6A interaction. (A) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1, pEGFP-C1-SC09 M2, pEGFP-C1-SC09 M2EDTM, or pEGFP-C1-SC09 M2CT into HEK293T cells for 48 h before preparation for cell lysates. Following immunoprecipitation with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody and a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (B and C) Plasmids expressing TRAPPC6A-myc and Flag-SC09M2 or Flag-SC09M2 with different amino acid deletions in the C terminus were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: A leucine residue at position 96 of M2 is required for the TRAPPC6A interaction. (A) pCAGGS-TRAPPC6A-myc was cotransfected with pEGFP-C1, pEGFP-C1-SC09 M2, pEGFP-C1-SC09 M2EDTM, or pEGFP-C1-SC09 M2CT into HEK293T cells for 48 h before preparation for cell lysates. Following immunoprecipitation with a mouse anti-GFP MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-GFP polyclonal antibody and a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively. (B and C) Plasmids expressing TRAPPC6A-myc and Flag-SC09M2 or Flag-SC09M2 with different amino acid deletions in the C terminus were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Immunoprecipitation, Western Blot, Expressing

    Dynamics of the interaction of M2 and TRAPPC6AΔ in wt and mutant WSN virus-infected cells. A549 cells were infected with wt influenza virus WSN (A), or one of the M2 deletion mutants WSN M2Del1 (B) and WSN M2Del2 (C), at an MOI of 5. At 4, 6, 8, 10, and 14 h p.i., the infected cells were fixed and stained with mouse anti-M2 MAb 14C2 and rabbit anti-TRAPPC6A polyclonal antibody, followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Dynamics of the interaction of M2 and TRAPPC6AΔ in wt and mutant WSN virus-infected cells. A549 cells were infected with wt influenza virus WSN (A), or one of the M2 deletion mutants WSN M2Del1 (B) and WSN M2Del2 (C), at an MOI of 5. At 4, 6, 8, 10, and 14 h p.i., the infected cells were fixed and stained with mouse anti-M2 MAb 14C2 and rabbit anti-TRAPPC6A polyclonal antibody, followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Mutagenesis, Infection, Staining, Incubation

    Mutation at position 96 of M2 affects its interaction with TRAPPC6A. (A) Sequence analysis of IAV M2 at position 96. All of the IAV M2 sequences deposited in GenBank by 6 July 2014 were downloaded. The identity of the amino acids at position 96 was statistically analyzed. (B) Plasmids expressing TRAPPC6A-Myc and Flag-WSNM2 or Flag-WSNM2 with different mutations at position 96 were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Mutation at position 96 of M2 affects its interaction with TRAPPC6A. (A) Sequence analysis of IAV M2 at position 96. All of the IAV M2 sequences deposited in GenBank by 6 July 2014 were downloaded. The identity of the amino acids at position 96 was statistically analyzed. (B) Plasmids expressing TRAPPC6A-Myc and Flag-WSNM2 or Flag-WSNM2 with different mutations at position 96 were cotransfected into HEK293T cells for 48 h before the preparation of cell lysates. Following immunoprecipitation with a mouse anti-Flag MAb, the immunoprecipitates were analyzed by Western blotting using a rabbit anti-Flag polyclonal antibody or a rabbit anti-Myc polyclonal antibody to reveal the presence of M2 and TRAPPC6A, respectively.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Mutagenesis, Sequencing, Expressing, Immunoprecipitation, Western Blot

    Effect of modulation of TRAPPC6AΔ expression on the cell surface expression of viral and cellular proteins. (A) A549 cells were transfected with siRNA targeting TRAPPC6AΔ or with nontargeting siRNA for 48 h and were then infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (B) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. Cells were fixed at 8 and 10 h p.i., left nonpermeabilized, and stained with the mouse anti-M2 MAb and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2 surface expression analysis by flow cytometry. The graph shows the fluorescence intensity of M2 surface expression. (C) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (D) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B. (E) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of HA was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (F) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of FGF2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (G) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. At 2 h p.i., the culture medium was replaced with medium supplemented with 25 μM amantadine. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Effect of modulation of TRAPPC6AΔ expression on the cell surface expression of viral and cellular proteins. (A) A549 cells were transfected with siRNA targeting TRAPPC6AΔ or with nontargeting siRNA for 48 h and were then infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (B) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. Cells were fixed at 8 and 10 h p.i., left nonpermeabilized, and stained with the mouse anti-M2 MAb and Alexa Fluor 488-conjugated donkey anti-mouse IgG(H+L) for M2 surface expression analysis by flow cytometry. The graph shows the fluorescence intensity of M2 surface expression. (C) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. Cell lysates were processed at 8 and 10 h p.i. and subjected to Western blotting using a mouse anti-M2 MAb to detect the expression level of M2. (D) The TRAPPC6AΔ-overexpressing A549 cell line or the A549 control cell line transduced with an empty retrovirus was infected with the WSN virus at an MOI of 3. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B. (E) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of HA was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-HA polyclonal antibody and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (F) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. The cell surface expression of FGF2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B by using the rabbit anti-FGF2 MAb and Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L). (G) A549 cells were treated with siRNA and infected with the WSN virus as described above for panel A. At 2 h p.i., the culture medium was replaced with medium supplemented with 25 μM amantadine. The cell surface expression of M2 was analyzed by flow cytometry at 8 and 10 h p.i. as described above for panel B.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Expressing, Transfection, Infection, Western Blot, Staining, Flow Cytometry, Cytometry, Fluorescence, Transduction

    TRAPPC6AΔ positively modulates influenza virus infection. (A) Endogenous expression of TRAPPC6AΔ in A549 cells. Whole lysates of A549 cells grown in 12-well plates were subjected to Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. HEK293T cell lysates transiently transfected with pCAGGS-TRAPPC6A or pCAGGS-TRAPPC6AΔ were used as a control. (B) siRNA knockdown of TRAPPC6AΔ in A549 cells. A549 cells were transfected with siRNA targeting TRAPPC6AΔ or nontargeting siRNA for 48 h. Whole-cell lysates were then collected and analyzed by Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. (C) Cell viability of siRNA-treated A549 cells measured by using the CellTiter-Glo assay. A549 cells were transfected with siRNA as described above for panel B. The data are presented as means ± standard deviations for triplicate transfections. (D) Virus replication in siRNA-treated A549 cells. Cells transfected with siRNA as described above for panel B were infected with WSN virus. At 24 and 48 h p.i., supernatants were collected and titrated for infectious virus by plaque assays in MDCK cells. Three independent experiments were performed, and data are shown as means ± standard deviations for triplicates from a representative experiment. **, P

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: TRAPPC6AΔ positively modulates influenza virus infection. (A) Endogenous expression of TRAPPC6AΔ in A549 cells. Whole lysates of A549 cells grown in 12-well plates were subjected to Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. HEK293T cell lysates transiently transfected with pCAGGS-TRAPPC6A or pCAGGS-TRAPPC6AΔ were used as a control. (B) siRNA knockdown of TRAPPC6AΔ in A549 cells. A549 cells were transfected with siRNA targeting TRAPPC6AΔ or nontargeting siRNA for 48 h. Whole-cell lysates were then collected and analyzed by Western blotting with a rabbit anti-TRAPPC6A polyclonal antibody. (C) Cell viability of siRNA-treated A549 cells measured by using the CellTiter-Glo assay. A549 cells were transfected with siRNA as described above for panel B. The data are presented as means ± standard deviations for triplicate transfections. (D) Virus replication in siRNA-treated A549 cells. Cells transfected with siRNA as described above for panel B were infected with WSN virus. At 24 and 48 h p.i., supernatants were collected and titrated for infectious virus by plaque assays in MDCK cells. Three independent experiments were performed, and data are shown as means ± standard deviations for triplicates from a representative experiment. **, P

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Infection, Expressing, Western Blot, Transfection, Glo Assay

    Confocal microscopy of WSN virus-infected cells stained for the Golgi apparatus or lysosomes. A549 cells were infected with the wt WSN virus at an MOI of 5. At the indicated time points, infected cells were fixed and stained with mouse anti-Giantin MAb and rabbit anti-M2 polyclonal antibody (A), mouse anti-LAMP1 MAb and rabbit anti-M2 polyclonal antibody (B), or mouse anti-LAMP1 MAb and rabbit anti-TRAPPC6A polyclonal antibody (C), followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Journal: Journal of Virology

    Article Title: Host Cellular Protein TRAPPC6AΔ Interacts with Influenza A Virus M2 Protein and Regulates Viral Propagation by Modulating M2 Trafficking

    doi: 10.1128/JVI.01757-16

    Figure Lengend Snippet: Confocal microscopy of WSN virus-infected cells stained for the Golgi apparatus or lysosomes. A549 cells were infected with the wt WSN virus at an MOI of 5. At the indicated time points, infected cells were fixed and stained with mouse anti-Giantin MAb and rabbit anti-M2 polyclonal antibody (A), mouse anti-LAMP1 MAb and rabbit anti-M2 polyclonal antibody (B), or mouse anti-LAMP1 MAb and rabbit anti-TRAPPC6A polyclonal antibody (C), followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG(H+L) (green) and Alexa Fluor 546 donkey anti-rabbit IgG(H+L) (red). Nuclei were stained with DAPI.

    Article Snippet: The following primary antibodies were obtained from commercial sources: rabbit anti-Flag polyclonal antibody (F7425; Sigma-Aldrich), mouse anti-Flag monoclonal antibody (F3165; Sigma-Aldrich), rabbit anti-Myc polyclonal antibody (C3965; Sigma-Aldrich), mouse anti-Myc monoclonal antibody (M4439; Sigma-Aldrich), mouse anti-actin monoclonal antibody (sc-47778; Santa Cruz, Dallas, TX), rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibody (10494-1-AP; Proteintech, Chicago, IL), rabbit anti-GFP polyclonal antibody (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP monoclonal antibody (ab1218; Abcam, Cambridge, MA), mouse anti-M2 monoclonal antibody (ab5416; Abcam), rabbit anti-M2 polyclonal antibody (GTX125951; GeneTex, Irvine, CA), rabbit anti-HA polyclonal antibody (11692-T54; Sino Biological Inc., Beijing, China), rabbit anti-FGF2 monoclonal antibody (ab92337; Abcam), mouse anti-LAMP1 monoclonal antibody (ab25630; Abcam), and mouse anti-Giantin monoclonal antibody (ab37266; Abcam).

    Techniques: Confocal Microscopy, Infection, Staining, Incubation

    Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following MEX4108 infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Validated miRNA targets are involved in immune response and cell proliferation in the lungs. Expression levels of (A) KRAS , BLIMP1 , and CDK6 (targets of let-7f); (B) SIRT1 , ZAP70 , and MYC (targets of miR-34c); (C) CAMTA1 (target of miR-129); and (D) PTEN (target of miR-18b) in BAL from rhesus macaques (five aged and three young adults) following MEX4108 infection are shown. Expression of mRNA was normalized to expression of RPL32 . Changes in gene expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. (A–D) *let-7f, miR-34c, miR-129, miR-18-b, KRAS , SIRT1 , CAMTA1 , and PTEN ; ‡ KRAS and ZAP70 ; † CDK6 and MYC . * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection

    Differentially expressed miRNAs in PBMCs following MEX4108. Expression levels of miR-18b (A) , miR-20a (B) , miR-192 (C) , miR-451 (D) , miR-138 (E) , miR-193b (F) , miR-132 (G) , let-7f (H) , and miR-146b (I) in PBMCs from rhesus macaques ( n = 11; five aged and six young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Differentially expressed miRNAs in PBMCs following MEX4108. Expression levels of miR-18b (A) , miR-20a (B) , miR-192 (C) , miR-451 (D) , miR-138 (E) , miR-193b (F) , miR-132 (G) , let-7f (H) , and miR-146b (I) in PBMCs from rhesus macaques ( n = 11; five aged and six young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection, Quantitative RT-PCR

    MEX4108 infection results in increased frequency of pDCs and robust production of cytokine, chemokine, and growth factor levels in the lungs. (A) The frequencies of DCs (lin − CD14 − HLA-DR + ) and macrophages/monocytes (lin − CD14 + HLA-DR − ) in BAL cells were measured by flow cytometry. (B) The frequencies of mDC (CD123 − CD11c + ) and pDC (CD123 + CD11c − ) in BAL cells were measured by flow cytometry. Longitudinal analyses of the frequency of immune cells within BAL were carried out using a one-way repeated-measures ANOVA model, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: MEX4108 infection results in increased frequency of pDCs and robust production of cytokine, chemokine, and growth factor levels in the lungs. (A) The frequencies of DCs (lin − CD14 − HLA-DR + ) and macrophages/monocytes (lin − CD14 + HLA-DR − ) in BAL cells were measured by flow cytometry. (B) The frequencies of mDC (CD123 − CD11c + ) and pDC (CD123 + CD11c − ) in BAL cells were measured by flow cytometry. Longitudinal analyses of the frequency of immune cells within BAL were carried out using a one-way repeated-measures ANOVA model, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Infection, Flow Cytometry, Cytometry

    Model of differentially expressed microRNAs, gene targets, and immune response in the lungs following MEX4108 infection.

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Model of differentially expressed microRNAs, gene targets, and immune response in the lungs following MEX4108 infection.

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Infection

    Pandemic H1N1 virus replicates to similar levels in young and aged macaques. Viral loads were measured using qRT-PCR using primers and probes specific for MEX4108 hemagglutinin (HA) in throat swabs (A) , nasal swabs (B) , ocular swabs (C) , and BAL fluid (D) . Viral genome copy number data were log transformed with base 10 and longitudinal changes of viral genome copy number between aged and young adults were compared using a two-way ANOVA, followed by Bonferroni's multiple comparison post-test to determine differences in viral load. Longitudinal changes were compared using repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values, mean ± SEM are shown. (A–D) * for aged animals; ‡ for young adult animals; *** ,‡‡ p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Pandemic H1N1 virus replicates to similar levels in young and aged macaques. Viral loads were measured using qRT-PCR using primers and probes specific for MEX4108 hemagglutinin (HA) in throat swabs (A) , nasal swabs (B) , ocular swabs (C) , and BAL fluid (D) . Viral genome copy number data were log transformed with base 10 and longitudinal changes of viral genome copy number between aged and young adults were compared using a two-way ANOVA, followed by Bonferroni's multiple comparison post-test to determine differences in viral load. Longitudinal changes were compared using repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test to explore differences between days postinfection and baseline (day 0) values, mean ± SEM are shown. (A–D) * for aged animals; ‡ for young adult animals; *** ,‡‡ p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Quantitative RT-PCR, Transformation Assay

    Differentially expressed miRNAs in BAL cells following MEX4108. Expression levels of let-7f (A) , miR-34c (B) , miR-129 (C) , miR-18b (D) , miR-146b (E) , miR-132 (F) , miR-192 (G) , and miR-138 (H) in BAL cells from rhesus macaques ( n = 8; five aged and three young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Journal: Viral Immunology

    Article Title: microRNAs Regulate Host Immune Response and Pathogenesis During Influenza Infection in Rhesus Macaques

    doi: 10.1089/vim.2015.0074

    Figure Lengend Snippet: Differentially expressed miRNAs in BAL cells following MEX4108. Expression levels of let-7f (A) , miR-34c (B) , miR-129 (C) , miR-18b (D) , miR-146b (E) , miR-132 (F) , miR-192 (G) , and miR-138 (H) in BAL cells from rhesus macaques ( n = 8; five aged and three young adults) following MEX4108 infection were determined using qRT-PCR. Expression of miRNAs was normalized to expression of U6 snRNA. Changes in microRNA expression postinfection were determined using one-way repeated-measures ANOVA, followed by Dunnett's multiple comparison post-test. Mean ± SEM are shown. * p

    Article Snippet: IgG and IgA binding antibody titers were measured in plasma and BAL supernatant by enzyme-linked immunosorbent assay (ELISA) using plates coated with 1 μg/mL recombinant MEX4108 HA protein (plasma) overnight at 4°C (Sino Biological, Inc., Beijing, China).

    Techniques: Expressing, Infection, Quantitative RT-PCR

    Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN -γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN - γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Mouse Assay, Expressing

    Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Influenza A H1N1 virus T-cell epitope screen in HLA-DQ6.2 mice, and Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. Spleen cells from Pandemrix-immunized HLA-DQ6.2 mice were stimulated in culture with pools of five overlapping 15-mer peptides each, covering a hemagglutinin (HA), b neuraminidase (NA) or c nucleoprotein (NP) from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus used in Pandemrix (pooling cells from 2 × 2 mice, n = 2). Recombinant hemagglutinin (rHA) and nucleoprotein (rNP; A/Puerto Rico/8/34 corresponding to the Pandemrix vaccine strain) were used as positive controls. d – f PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals (sleep clinic patients without a diagnosis of NT1) were stimulated in culture with single 15-mer peptides, derived from the same vaccine virus strain ( n = 1–2). The expression of IFN-γ or IL-2 was measured by FMIA (protein) or RT-qPCR (mRNA). Results are expressed as ratios between cytokine concentrations (lines representing means) or relative gene expressions (dots representing single values, bars representing means) measured in peptide-stimulated and negative control samples (stimulation index). An asterisk (*) indicates a pool or single peptide that was selected for further testing.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Mouse Assay, Recombinant, Derivative Assay, Expressing, Quantitative RT-PCR, Negative Control

    Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Mapping of identified influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. a , d PBMC from pediatric Pandemrix-associated NT1 patients (NT1; validation cohort) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with overlapping 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus neuraminidase (NA) or nucleoprotein (NP), as indicated. b , c , e , f PBMC from NT1 patients (invariably HLA-DQB1*0602 positive; discovery and validation cohorts combined; HLA-DQB1*0602 homozygous NT1 patients marked with red dots) or HLA-DQB1*0602 positive (C/DQ6+) or negative (C/DQ6−) healthy controls were stimulated with single NA- or NP-derived peptides. The secretion of IFN-γ was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Derivative Assay, Negative Control

    Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Journal: Nature Communications

    Article Title: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

    doi: 10.1038/s41467-021-22637-8

    Figure Lengend Snippet: Identification of influenza A H1N1 virus T-cell epitopes in Pandemrix-associated NT1 patients. PBMC from pediatric Pandemrix-associated NT1 patients (NT1) or pediatric Pandemrix-vaccinated healthy controls (C) were stimulated in culture with single 15-mer peptides from influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) vaccine virus hemagglutinin (HA), neuraminidase (NA) or nucleoprotein (NP) (discovery cohort). Recombinant neuraminidase (rNA) and nucleoprotein (rNP) were used as positive controls. The secretion of IFN-γ ( a , c , e ) or IL-2 ( b , d , f ) was measured by FMIA (protein). Results are expressed as the ratio between cytokine concentrations measured in peptide-stimulated and negative control samples (stimulation index). Statistical comparisons between groups were performed, using Kruskal–Wallis and Dunn’s multiple comparisons tests.

    Article Snippet: Peptides and recombinant proteins15-mer peptides covering hemagglutinin (GenBank entry ACP41953.1) and neuraminidase (YP_009118627.1) of influenza (A/California/07/2009 (H1N1)), and nucleoprotein (ADE29096.1) of influenza (A/reassortant/NYMC X-179A (California/07/2009 × NYMC X-157)(H1N1)) with 12 amino acid overlap were produced.

    Techniques: Recombinant, Negative Control