lasv gp Search Results


lasv gp josiah  (NewLink Genetics)
90
NewLink Genetics lasv gp josiah
<t>LASV</t> pseudovirion entry is TIM-1 dependent in Vero cells. (A) Cell surface detection of endogenous DG expression on Vero <t>or</t> <t>HEK</t> 293T cells. Live cells were stained with polyclonal DG antisera (top) or the mannosylation-dependent anti-αDG mAb IIH6 (bottom). Filled histograms represent cells stained with isotype control antisera or mAb; unfilled histograms represent cells stained with anti-DG polyclonal antisera or IIH6. (B) Cell surface expression of endogenous TIM-1, Axl, Mer, Tyro3, or TIM-4 on WT Vero cells. Also shown are TIM-1 and Axl staining of Axl-KO, TIM-KO1, TIM-KO2, or Axl/TIM-KO (DKO) Vero cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-TIM-1, anti-Axl, anti-Mer, anti-Tyro3, or anti-TIM-4 polyclonal antibody. (C and D) LASV-VSV (C) or adenovirus-eGFP (D) transduction of WT, TIM-1-KO, Axl-KO, or DKO Vero cells. Serial dilutions of virus were applied to each cell type in duplicate. Transduction is represented as the percentage of cells that are eGFP positive, as assessed by flow cytometry. Data points are representative of results from three independent experiments with the indicated standard errors of the means (SEM). (E) LASV-LCMV transduction (MOI = ∼0.2) of WT, TIM-1-KO, Axl-KO, or DKO Vero cell lines. Data points are representative of results from three independent experiments with the indicated SEM. (F and G) Ability of adenovirus vector (Ad)-delivered TIM-1 to enhance transduction of LASV-VSV in WT, TIM-1-KO, or TIM-1- and Axl-KO (DKO) Vero cells. Forty-eight hours following Ad transduction, cells were divided and assessed independently for TIM-1 expression (F) or LASV-VSV transduction (G). Bars in each panel represent results from one of three independent experiments. (F) Cell surface expression of TIM-1 48 h following transduction of WT or KO Vero cells with Ad-TIM-1 (MOI of 2, 4, or 8). Cells were surface stained with 2 μg/ml anti-TIM-1 polyclonal antibodies. TIM-1 expression is represented as geometric mean fluorescence index (MFI) values. (G) LASV-VSV transductions of empty (E) or TIM-1-expressing (T) adenovirus vector-transduced WT or KO Vero cells. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells, as assessed by flow cytometry.
Lasv Gp Josiah, supplied by NewLink Genetics, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/lasv+gp/lasv+gp++josiah+/pmc06069209-358-14-23
Average 90 stars, based on 1 article reviews
lasv gp josiah - by Bioz Stars, 2026-10
90/100 stars
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lasv gp  (GenScript corporation)
90
GenScript corporation lasv gp
Recombinant <t>LASV</t> <t>GP</t> protein expressed in Drosophila S2 cells after purification using IAC with R3621 mAb followed by SEC. Western blot of purified LASV GP. Four μg of the vaccine antigen was probed with mAb 10.4B, 19.7E, 2B12, and 37.7H targeting epitopes on GP1, GP2, and GPC, at a dilution of 1:1000.
Lasv Gp, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/lasv+gp/lasv+gp/pmc10081732-100-60-62
Average 90 stars, based on 1 article reviews
lasv gp - by Bioz Stars, 2026-10
90/100 stars
  Buy from Supplier
anti-lasv gp polyclonal antibody  (Creative BioMart)
N/A
2-3 weeks +4°C, -20°C long termhttp://www.creative-diagnostics.com/LASV-GP-antibody-243612-147.htm
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Image Search Results


LASV pseudovirion entry is TIM-1 dependent in Vero cells. (A) Cell surface detection of endogenous DG expression on Vero or HEK 293T cells. Live cells were stained with polyclonal DG antisera (top) or the mannosylation-dependent anti-αDG mAb IIH6 (bottom). Filled histograms represent cells stained with isotype control antisera or mAb; unfilled histograms represent cells stained with anti-DG polyclonal antisera or IIH6. (B) Cell surface expression of endogenous TIM-1, Axl, Mer, Tyro3, or TIM-4 on WT Vero cells. Also shown are TIM-1 and Axl staining of Axl-KO, TIM-KO1, TIM-KO2, or Axl/TIM-KO (DKO) Vero cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-TIM-1, anti-Axl, anti-Mer, anti-Tyro3, or anti-TIM-4 polyclonal antibody. (C and D) LASV-VSV (C) or adenovirus-eGFP (D) transduction of WT, TIM-1-KO, Axl-KO, or DKO Vero cells. Serial dilutions of virus were applied to each cell type in duplicate. Transduction is represented as the percentage of cells that are eGFP positive, as assessed by flow cytometry. Data points are representative of results from three independent experiments with the indicated standard errors of the means (SEM). (E) LASV-LCMV transduction (MOI = ∼0.2) of WT, TIM-1-KO, Axl-KO, or DKO Vero cell lines. Data points are representative of results from three independent experiments with the indicated SEM. (F and G) Ability of adenovirus vector (Ad)-delivered TIM-1 to enhance transduction of LASV-VSV in WT, TIM-1-KO, or TIM-1- and Axl-KO (DKO) Vero cells. Forty-eight hours following Ad transduction, cells were divided and assessed independently for TIM-1 expression (F) or LASV-VSV transduction (G). Bars in each panel represent results from one of three independent experiments. (F) Cell surface expression of TIM-1 48 h following transduction of WT or KO Vero cells with Ad-TIM-1 (MOI of 2, 4, or 8). Cells were surface stained with 2 μg/ml anti-TIM-1 polyclonal antibodies. TIM-1 expression is represented as geometric mean fluorescence index (MFI) values. (G) LASV-VSV transductions of empty (E) or TIM-1-expressing (T) adenovirus vector-transduced WT or KO Vero cells. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells, as assessed by flow cytometry.

Journal: Journal of Virology

Article Title: TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

doi: 10.1128/JVI.00093-18

Figure Lengend Snippet: LASV pseudovirion entry is TIM-1 dependent in Vero cells. (A) Cell surface detection of endogenous DG expression on Vero or HEK 293T cells. Live cells were stained with polyclonal DG antisera (top) or the mannosylation-dependent anti-αDG mAb IIH6 (bottom). Filled histograms represent cells stained with isotype control antisera or mAb; unfilled histograms represent cells stained with anti-DG polyclonal antisera or IIH6. (B) Cell surface expression of endogenous TIM-1, Axl, Mer, Tyro3, or TIM-4 on WT Vero cells. Also shown are TIM-1 and Axl staining of Axl-KO, TIM-KO1, TIM-KO2, or Axl/TIM-KO (DKO) Vero cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-TIM-1, anti-Axl, anti-Mer, anti-Tyro3, or anti-TIM-4 polyclonal antibody. (C and D) LASV-VSV (C) or adenovirus-eGFP (D) transduction of WT, TIM-1-KO, Axl-KO, or DKO Vero cells. Serial dilutions of virus were applied to each cell type in duplicate. Transduction is represented as the percentage of cells that are eGFP positive, as assessed by flow cytometry. Data points are representative of results from three independent experiments with the indicated standard errors of the means (SEM). (E) LASV-LCMV transduction (MOI = ∼0.2) of WT, TIM-1-KO, Axl-KO, or DKO Vero cell lines. Data points are representative of results from three independent experiments with the indicated SEM. (F and G) Ability of adenovirus vector (Ad)-delivered TIM-1 to enhance transduction of LASV-VSV in WT, TIM-1-KO, or TIM-1- and Axl-KO (DKO) Vero cells. Forty-eight hours following Ad transduction, cells were divided and assessed independently for TIM-1 expression (F) or LASV-VSV transduction (G). Bars in each panel represent results from one of three independent experiments. (F) Cell surface expression of TIM-1 48 h following transduction of WT or KO Vero cells with Ad-TIM-1 (MOI of 2, 4, or 8). Cells were surface stained with 2 μg/ml anti-TIM-1 polyclonal antibodies. TIM-1 expression is represented as geometric mean fluorescence index (MFI) values. (G) LASV-VSV transductions of empty (E) or TIM-1-expressing (T) adenovirus vector-transduced WT or KO Vero cells. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells, as assessed by flow cytometry.

Article Snippet: VSV pseudovirions were produced by transfecting HEK 293T cells with plasmids containing either the LASV GP (Josiah; a kind gift from Robert Mandell, NewLink Genetics), VSV G (Indiana; a kind gift from Paul McCray, University of Iowa), or full-length EBOV GP (Mayinga; a kind gift Robert Davey, Texas Biomedical Research Institute) gene 18 to 24 h prior to transduction with LASV GPC-pseudotyped VSVΔG-eGFP at a multiplicity of infection (MOI) of ∼1.

Techniques: Expressing, Staining, Control, Transduction, Virus, Flow Cytometry, Plasmid Preparation, Fluorescence

LASV-VSV entry is αDG dependent in HEK 293T cells. (A) Cell surface expression of endogenous DG on WT or DG-KO HEK 293T cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-DG polyclonal antibodies. (B and C) LASV-VSV (B) or VSV-VSV (C) transduction in WT or DG-KO HEK 293T cells. Serial dilutions of pseudovirus were applied to each cell type in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to 1 μl of pseudovirus in WT HEK 293T cells, as assessed by flow cytometry. Shown are means and standard errors of the means of data from three to four independent experiments. (D and E) Dose-dependent rescue of LASV-VSV transduction in αDG-transfected DG-KO HEK 293T cells. Cells were transfected with 0, 0.25, 0.5, or 1 μg DG-expressing plasmid DNA (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for the surface expression of DG (D) or LASV-VSV transduction (E). Shown are means and standard errors of the means of data from four independent experiments. (D) Cell surface expression of DG on DG-transfected DG-KO HEK 293T cells. Cells were stained with 2 μg/ml anti-DG polyclonal antisera, and values shown are mean fluorescence index (MFI) values. (E) LASV-VSV transduction of αDG-transfected DG-KO HEK 293T cells. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to untransfected cells, as assessed by flow cytometry. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Journal: Journal of Virology

Article Title: TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

doi: 10.1128/JVI.00093-18

Figure Lengend Snippet: LASV-VSV entry is αDG dependent in HEK 293T cells. (A) Cell surface expression of endogenous DG on WT or DG-KO HEK 293T cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-DG polyclonal antibodies. (B and C) LASV-VSV (B) or VSV-VSV (C) transduction in WT or DG-KO HEK 293T cells. Serial dilutions of pseudovirus were applied to each cell type in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to 1 μl of pseudovirus in WT HEK 293T cells, as assessed by flow cytometry. Shown are means and standard errors of the means of data from three to four independent experiments. (D and E) Dose-dependent rescue of LASV-VSV transduction in αDG-transfected DG-KO HEK 293T cells. Cells were transfected with 0, 0.25, 0.5, or 1 μg DG-expressing plasmid DNA (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for the surface expression of DG (D) or LASV-VSV transduction (E). Shown are means and standard errors of the means of data from four independent experiments. (D) Cell surface expression of DG on DG-transfected DG-KO HEK 293T cells. Cells were stained with 2 μg/ml anti-DG polyclonal antisera, and values shown are mean fluorescence index (MFI) values. (E) LASV-VSV transduction of αDG-transfected DG-KO HEK 293T cells. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to untransfected cells, as assessed by flow cytometry. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: VSV pseudovirions were produced by transfecting HEK 293T cells with plasmids containing either the LASV GP (Josiah; a kind gift from Robert Mandell, NewLink Genetics), VSV G (Indiana; a kind gift from Paul McCray, University of Iowa), or full-length EBOV GP (Mayinga; a kind gift Robert Davey, Texas Biomedical Research Institute) gene 18 to 24 h prior to transduction with LASV GPC-pseudotyped VSVΔG-eGFP at a multiplicity of infection (MOI) of ∼1.

Techniques: Expressing, Staining, Control, Transduction, Flow Cytometry, Transfection, Plasmid Preparation, Fluorescence

TIM-1 serves as a receptor for LASV-VSV in the absence of αDG. (A) Cell surface expression of endogenous TIM-1 or Axl expression on WT HEK 293T cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-Axl or anti-TIM-1 polyclonal antisera. Analyses of Axl and TIM-1 surface expression on HEK 293T cells were performed in parallel with ectopic Axl and TIM-1 surface staining of transfected cells, shown as MFI in panels below. (B to E) Dose-dependent rescue of LASV-VSV transduction in TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells. Each cell type was transfected with 0, 0.25, 0.5, or 1 μg TIM-1- or Axl-expressing plasmid DNA (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for TIM-1 or Axl expression (B and D) or LASV-VSV transduction (C and E). Bars in each panel are representative of data from four independent experiments with the indicated SEM. (B and D) Cell surface expression analysis of TIM-1 or Axl expression on TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells, respectively. Cells were stained with 2 μg/ml anti-TIM-1 or anti-Axl polyclonal antibodies. TIM-1 or Axl expression is represented as units of geometric means. (C and E) LASV-VSV transduction of TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells, respectively. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to untransfected cells, as assessed by flow cytometry. Shown are means and standard errors of the means of results from four independent experiments. (F) Axl expression in DG-KO HEK 293T cells enhances Zika virus infection. DG-KO cells were transfected with an empty vector or a vector expressing Axl. Zika virus (Puerto Rico strain) (MOI of ∼0.25) was added to Axl or empty plasmid-transfected DG-KO cells. Infected cells were lysed at day 3 of infection, and viral genome equivalents were determined by qRT-PCR. (G) HEK 293T cells transfected with empty, Axl-expressing, or murine BST-2-expressing plasmids were plated on the insert of a transwell in serum-free medium. Medium containing 10% FCS with or without 0.5 μg/ml of Gas6 was added to the bottom of the transwell apparatus. Cell migration to the bottom of the transwell filter was determined 24 h later. Data shown represent results of one of two studies that were performed in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Journal: Journal of Virology

Article Title: TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

doi: 10.1128/JVI.00093-18

Figure Lengend Snippet: TIM-1 serves as a receptor for LASV-VSV in the absence of αDG. (A) Cell surface expression of endogenous TIM-1 or Axl expression on WT HEK 293T cells. Filled histograms represent cells stained with the isotype control; unfilled histograms represent cells stained with 2 μg/ml anti-Axl or anti-TIM-1 polyclonal antisera. Analyses of Axl and TIM-1 surface expression on HEK 293T cells were performed in parallel with ectopic Axl and TIM-1 surface staining of transfected cells, shown as MFI in panels below. (B to E) Dose-dependent rescue of LASV-VSV transduction in TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells. Each cell type was transfected with 0, 0.25, 0.5, or 1 μg TIM-1- or Axl-expressing plasmid DNA (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for TIM-1 or Axl expression (B and D) or LASV-VSV transduction (C and E). Bars in each panel are representative of data from four independent experiments with the indicated SEM. (B and D) Cell surface expression analysis of TIM-1 or Axl expression on TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells, respectively. Cells were stained with 2 μg/ml anti-TIM-1 or anti-Axl polyclonal antibodies. TIM-1 or Axl expression is represented as units of geometric means. (C and E) LASV-VSV transduction of TIM-1- or Axl-transfected WT or DG-KO HEK 293T cells, respectively. Equivalent volumes of LASV-VSV were applied to cells in duplicate. Transduction is represented as the percentage of eGFP-positive cells relative to untransfected cells, as assessed by flow cytometry. Shown are means and standard errors of the means of results from four independent experiments. (F) Axl expression in DG-KO HEK 293T cells enhances Zika virus infection. DG-KO cells were transfected with an empty vector or a vector expressing Axl. Zika virus (Puerto Rico strain) (MOI of ∼0.25) was added to Axl or empty plasmid-transfected DG-KO cells. Infected cells were lysed at day 3 of infection, and viral genome equivalents were determined by qRT-PCR. (G) HEK 293T cells transfected with empty, Axl-expressing, or murine BST-2-expressing plasmids were plated on the insert of a transwell in serum-free medium. Medium containing 10% FCS with or without 0.5 μg/ml of Gas6 was added to the bottom of the transwell apparatus. Cell migration to the bottom of the transwell filter was determined 24 h later. Data shown represent results of one of two studies that were performed in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: VSV pseudovirions were produced by transfecting HEK 293T cells with plasmids containing either the LASV GP (Josiah; a kind gift from Robert Mandell, NewLink Genetics), VSV G (Indiana; a kind gift from Paul McCray, University of Iowa), or full-length EBOV GP (Mayinga; a kind gift Robert Davey, Texas Biomedical Research Institute) gene 18 to 24 h prior to transduction with LASV GPC-pseudotyped VSVΔG-eGFP at a multiplicity of infection (MOI) of ∼1.

Techniques: Expressing, Staining, Control, Transfection, Transduction, Plasmid Preparation, Flow Cytometry, Virus, Infection, Quantitative RT-PCR, Migration

TIM-1 mediates LASV pseudovirion transduction through a PtdSer-dependent mechanism. (A and B) Anti-human TIM-1 antibody ARD5 inhibits TIM-1-dependent LASV pseudovirion transduction in HEK 293T cells in a dose-dependent manner. (A) WT or DG-KO HEK 293T cells transfected with a TIM-1-expressing plasmid were incubated with serial dilutions of ARD5 prior to the addition of LASV-VSV. Relative transduction is represented as the percentage of eGFP-positive cells relative to cells incubated without ARD5, as assessed by flow cytometry. (B) WT or Axl-KO Vero cells were incubated with serial dilutions of ARD5 prior to the addition of LASV-LCMV, and data are shown as relative transduction, as described above. Data points are representative of results from three independent experiments with the indicated SEM. (C to F) Ability of two human TIM-1 mutants to support LASV-VSV transduction in DG-KO HEK 293T cells. Plasmids expressing a PtdSer pocket mutant, TIM-1 (ND115DN), or a chimeric TIM-1 that has the MLD replaced with the proline-rich region (PRR) of murine leukemia virus glycoprotein were transfected into WT or DG-KO cells. Cells were transfected with 0, 0.25, 0.5, or 1 μg ND115DN or PRR plasmid (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for TIM-1 expression (C and E) or LASV-VSV transduction (D and F). Equivalent volumes of LASV-VSV were applied to cells in duplicate in transduction studies. Shown are fold change values relative to transduction levels of untransfected cells of the same type, as assessed by flow cytometry. Shown in panels A and B are means and standard errors of the means of data from three independent experiments. Shown in panels C to F are means and standard errors of the means of data from 3 to 5 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Journal: Journal of Virology

Article Title: TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

doi: 10.1128/JVI.00093-18

Figure Lengend Snippet: TIM-1 mediates LASV pseudovirion transduction through a PtdSer-dependent mechanism. (A and B) Anti-human TIM-1 antibody ARD5 inhibits TIM-1-dependent LASV pseudovirion transduction in HEK 293T cells in a dose-dependent manner. (A) WT or DG-KO HEK 293T cells transfected with a TIM-1-expressing plasmid were incubated with serial dilutions of ARD5 prior to the addition of LASV-VSV. Relative transduction is represented as the percentage of eGFP-positive cells relative to cells incubated without ARD5, as assessed by flow cytometry. (B) WT or Axl-KO Vero cells were incubated with serial dilutions of ARD5 prior to the addition of LASV-LCMV, and data are shown as relative transduction, as described above. Data points are representative of results from three independent experiments with the indicated SEM. (C to F) Ability of two human TIM-1 mutants to support LASV-VSV transduction in DG-KO HEK 293T cells. Plasmids expressing a PtdSer pocket mutant, TIM-1 (ND115DN), or a chimeric TIM-1 that has the MLD replaced with the proline-rich region (PRR) of murine leukemia virus glycoprotein were transfected into WT or DG-KO cells. Cells were transfected with 0, 0.25, 0.5, or 1 μg ND115DN or PRR plasmid (increase in DNA is indicated with unfilled triangles). After 48 h, cells were divided and assessed independently for TIM-1 expression (C and E) or LASV-VSV transduction (D and F). Equivalent volumes of LASV-VSV were applied to cells in duplicate in transduction studies. Shown are fold change values relative to transduction levels of untransfected cells of the same type, as assessed by flow cytometry. Shown in panels A and B are means and standard errors of the means of data from three independent experiments. Shown in panels C to F are means and standard errors of the means of data from 3 to 5 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: VSV pseudovirions were produced by transfecting HEK 293T cells with plasmids containing either the LASV GP (Josiah; a kind gift from Robert Mandell, NewLink Genetics), VSV G (Indiana; a kind gift from Paul McCray, University of Iowa), or full-length EBOV GP (Mayinga; a kind gift Robert Davey, Texas Biomedical Research Institute) gene 18 to 24 h prior to transduction with LASV GPC-pseudotyped VSVΔG-eGFP at a multiplicity of infection (MOI) of ∼1.

Techniques: Transduction, Transfection, Expressing, Plasmid Preparation, Incubation, Flow Cytometry, Mutagenesis, Virus

Recombinant LASV GP protein expressed in Drosophila S2 cells after purification using IAC with R3621 mAb followed by SEC. Western blot of purified LASV GP. Four μg of the vaccine antigen was probed with mAb 10.4B, 19.7E, 2B12, and 37.7H targeting epitopes on GP1, GP2, and GPC, at a dilution of 1:1000.

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Recombinant LASV GP protein expressed in Drosophila S2 cells after purification using IAC with R3621 mAb followed by SEC. Western blot of purified LASV GP. Four μg of the vaccine antigen was probed with mAb 10.4B, 19.7E, 2B12, and 37.7H targeting epitopes on GP1, GP2, and GPC, at a dilution of 1:1000.

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Recombinant, Purification, Western Blot

Total IgG and IgG subclasses generated in a 3-dose study of adjuvanted LASV GP in SW mice. (A) Vaccination schedule. (B) Experimental groups. (C) Geometric mean titers (GMT) with 95% CI of LASV GP-specific IgG MFI of serum samples from all groups of mice immunized with each formulation (n=8 for each group). Each symbol represents the mean of duplicates from individual mice. A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences in IgG titers between primary and subsequent doses. (D) GMT with 95% CI of LASV-GP specific IgG subclass MFI of pooled sera from experimental groups post-dose 3. Statistical significance was not calculated as each data point consisted of a single pooled sample ran in duplicates (*p < 0.05, **p < 0.01, ****p < 0.0001).

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Total IgG and IgG subclasses generated in a 3-dose study of adjuvanted LASV GP in SW mice. (A) Vaccination schedule. (B) Experimental groups. (C) Geometric mean titers (GMT) with 95% CI of LASV GP-specific IgG MFI of serum samples from all groups of mice immunized with each formulation (n=8 for each group). Each symbol represents the mean of duplicates from individual mice. A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences in IgG titers between primary and subsequent doses. (D) GMT with 95% CI of LASV-GP specific IgG subclass MFI of pooled sera from experimental groups post-dose 3. Statistical significance was not calculated as each data point consisted of a single pooled sample ran in duplicates (*p < 0.05, **p < 0.01, ****p < 0.0001).

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Generated, Formulation, Comparison

Total IgG and IgG subclass titers from a 2-dose study with 2 or 10 μg of LASV GP formulated with select adjuvants. (A) Vaccination schedule. (B) Experimental groups. (C) Geometric means (GMT) with 95% CI of LASV GP-specific IgG MFI of serum from all groups of mice immunized with each formulation. Mice were given either 2 or 10 μg of antigen formulated with adjuvants selected from . Each symbol represents the mean of duplicates from individual mice (n=7 or 8 for each group). A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences across antigen dose and adjuvant formulations (D) The median of individual MFIs plotted with 5-95 percentile of LASV GP-specific IgG subclasses from each mouse group. Statistical differences between titers of the same IgG subclass across vaccine formulations was calculated using a two-way ANOVA followed by Tukey’s multiple comparison (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Total IgG and IgG subclass titers from a 2-dose study with 2 or 10 μg of LASV GP formulated with select adjuvants. (A) Vaccination schedule. (B) Experimental groups. (C) Geometric means (GMT) with 95% CI of LASV GP-specific IgG MFI of serum from all groups of mice immunized with each formulation. Mice were given either 2 or 10 μg of antigen formulated with adjuvants selected from . Each symbol represents the mean of duplicates from individual mice (n=7 or 8 for each group). A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences across antigen dose and adjuvant formulations (D) The median of individual MFIs plotted with 5-95 percentile of LASV GP-specific IgG subclasses from each mouse group. Statistical differences between titers of the same IgG subclass across vaccine formulations was calculated using a two-way ANOVA followed by Tukey’s multiple comparison (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Formulation, Comparison, Adjuvant

IgG subclass ratios of immunized mice from experimental groups. (A) IgG2a/IgG1 and (B) IgG2b/IgG1 ratio. Geometric means (GMT) with 95% CI of LASV GP-specific IgG subclass MFI ratio of serum from all groups of mice immunized with each formulation from . PD2 except for LASV GP alone (PD3). Each symbol represents the mean of duplicates from individual mice (n=7 or 8 for each group). A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences in subclass ratios compared to LASV GP alone (*p < 0.05).

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: IgG subclass ratios of immunized mice from experimental groups. (A) IgG2a/IgG1 and (B) IgG2b/IgG1 ratio. Geometric means (GMT) with 95% CI of LASV GP-specific IgG subclass MFI ratio of serum from all groups of mice immunized with each formulation from . PD2 except for LASV GP alone (PD3). Each symbol represents the mean of duplicates from individual mice (n=7 or 8 for each group). A two-way ANOVA followed by a Tukey’s multiple comparison was used to determine statistical differences in subclass ratios compared to LASV GP alone (*p < 0.05).

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Formulation, Comparison

Cytokine secretion measured from mouse splenocytes using a FluoroSpot assay. Geometric mean (GMT) with 95% confidence intervals of (A) IL-2, (B) TNF-α, (C) IFN-γ and (D) polyfunctional cytokine secretion from splenocytes harvested from mouse groups one-week post-dose 2 and stimulated with a LASV GP peptide pool for 24 hours. (n=3 for the ISA 720 groups, n=1-2 for LiteVax groups, and n=3-4 for GPI-0100 groups) A two-way ANOVA, followed by a Tukey’s multiple comparison was used to compare the number of cytokine-secreting cells from immunized groups to the group receiving LASV GP alone, and between different adjuvant formulations of the same antigen concentration, respectively. (*p < 0.05, **p < 0.01).

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Cytokine secretion measured from mouse splenocytes using a FluoroSpot assay. Geometric mean (GMT) with 95% confidence intervals of (A) IL-2, (B) TNF-α, (C) IFN-γ and (D) polyfunctional cytokine secretion from splenocytes harvested from mouse groups one-week post-dose 2 and stimulated with a LASV GP peptide pool for 24 hours. (n=3 for the ISA 720 groups, n=1-2 for LiteVax groups, and n=3-4 for GPI-0100 groups) A two-way ANOVA, followed by a Tukey’s multiple comparison was used to compare the number of cytokine-secreting cells from immunized groups to the group receiving LASV GP alone, and between different adjuvant formulations of the same antigen concentration, respectively. (*p < 0.05, **p < 0.01).

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Flurospot, Comparison, Adjuvant, Concentration Assay

Cytokine secretion measured from mouse splenocytes using Multiplex Cytokine assay (A) IFN-γ, (B) TNF-α, (C) IL-2, (D) IL-4 and (E) IL-5 cytokine secretion from splenocytes harvested from mouse groups one-week post-dose 2 and stimulated with LASV GP antigen for 48 (blue) and 96 hours (gold). Bars indicate the GMT with 95% confidence intervals. (n=3 for the ISA 720 groups, n=2-3 for LiteVax groups, and n=3-4 for GPI-0100 groups) A two-way ANOVA, followed by a Tukey’s multiple comparison was used to compare the secreted cytokine concentrations from immunized groups compared to mice receiving LASV GP alone and between different adjuvant formulations of the same antigen concentration, respectively. (*p < 0.05, **p < 0.01, ***p < 0.001).

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Cytokine secretion measured from mouse splenocytes using Multiplex Cytokine assay (A) IFN-γ, (B) TNF-α, (C) IL-2, (D) IL-4 and (E) IL-5 cytokine secretion from splenocytes harvested from mouse groups one-week post-dose 2 and stimulated with LASV GP antigen for 48 (blue) and 96 hours (gold). Bars indicate the GMT with 95% confidence intervals. (n=3 for the ISA 720 groups, n=2-3 for LiteVax groups, and n=3-4 for GPI-0100 groups) A two-way ANOVA, followed by a Tukey’s multiple comparison was used to compare the secreted cytokine concentrations from immunized groups compared to mice receiving LASV GP alone and between different adjuvant formulations of the same antigen concentration, respectively. (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).

Techniques: Multiplex Assay, Cytokine Assay, Comparison, Adjuvant, Concentration Assay