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full length ca09 ha  (Sino Biological)


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    Sino Biological full length ca09 ha
    Initial immune characterization of HA DNA-LNP formulations and immunogenicity (A) Schematic of DNA-LNP N/P ratios and immunization regimen. (B–D) Biophysical characterization of DNA-LNPs at different N/P ratios. (B) Particle size; (C) polydispersity index (PDI); (D) zeta potential. (E) Representative fluorescence-activated cell sorting (FACS) plots of GC B cells. (F) Bar plots quantifying frequency of GC B cells. (G) Frequency of <t>CA09</t> HA-specific GC B cells. (H) Frequency of activated Tfh cells. (I) IFNγ ELISpot of splenocytes. (J) Representative TEM images of HA DNA-LNP (top) and HA mRNA-LNP (bottom). Scale bar 100 nm (K and L) Fold change cytokine induction in DLNs at 4 h (K) and 24 h (L) after immunization quantified using Luminex. (M and N) ELISpot assay measuring IFNα (M) and IFNγ (N) 20 h after stimulation of splenocytes ex vivo with DNA-LNP, plasmid DNA, or DNA-LNP in the presence of chemical inhibitors to the indicated DNA sensors. (O) Schematic of relevant pathways implicated in DNA-LNP sensing. Dots represent individual animals; n = 8–9 (E–H), n = 5 (I, K, and L), or n = 3–4 animals per group (L and M); data pooled or representative from two independent experiments (E–H, M, and N) or from one independent experiment (I–L). Plots show mean with SD (B–D and I) or geometric mean with geometric SD (F–H and K–N). Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (F–H, K, and L) or compared to DNA-LNP control (M and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
    Full Length Ca09 Ha, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/full length ca09 ha/product/Sino Biological
    Average 94 stars, based on 12 article reviews
    full length ca09 ha - by Bioz Stars, 2026-03
    94/100 stars

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    1) Product Images from "Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity"

    Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2025.102035

    Initial immune characterization of HA DNA-LNP formulations and immunogenicity (A) Schematic of DNA-LNP N/P ratios and immunization regimen. (B–D) Biophysical characterization of DNA-LNPs at different N/P ratios. (B) Particle size; (C) polydispersity index (PDI); (D) zeta potential. (E) Representative fluorescence-activated cell sorting (FACS) plots of GC B cells. (F) Bar plots quantifying frequency of GC B cells. (G) Frequency of CA09 HA-specific GC B cells. (H) Frequency of activated Tfh cells. (I) IFNγ ELISpot of splenocytes. (J) Representative TEM images of HA DNA-LNP (top) and HA mRNA-LNP (bottom). Scale bar 100 nm (K and L) Fold change cytokine induction in DLNs at 4 h (K) and 24 h (L) after immunization quantified using Luminex. (M and N) ELISpot assay measuring IFNα (M) and IFNγ (N) 20 h after stimulation of splenocytes ex vivo with DNA-LNP, plasmid DNA, or DNA-LNP in the presence of chemical inhibitors to the indicated DNA sensors. (O) Schematic of relevant pathways implicated in DNA-LNP sensing. Dots represent individual animals; n = 8–9 (E–H), n = 5 (I, K, and L), or n = 3–4 animals per group (L and M); data pooled or representative from two independent experiments (E–H, M, and N) or from one independent experiment (I–L). Plots show mean with SD (B–D and I) or geometric mean with geometric SD (F–H and K–N). Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (F–H, K, and L) or compared to DNA-LNP control (M and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
    Figure Legend Snippet: Initial immune characterization of HA DNA-LNP formulations and immunogenicity (A) Schematic of DNA-LNP N/P ratios and immunization regimen. (B–D) Biophysical characterization of DNA-LNPs at different N/P ratios. (B) Particle size; (C) polydispersity index (PDI); (D) zeta potential. (E) Representative fluorescence-activated cell sorting (FACS) plots of GC B cells. (F) Bar plots quantifying frequency of GC B cells. (G) Frequency of CA09 HA-specific GC B cells. (H) Frequency of activated Tfh cells. (I) IFNγ ELISpot of splenocytes. (J) Representative TEM images of HA DNA-LNP (top) and HA mRNA-LNP (bottom). Scale bar 100 nm (K and L) Fold change cytokine induction in DLNs at 4 h (K) and 24 h (L) after immunization quantified using Luminex. (M and N) ELISpot assay measuring IFNα (M) and IFNγ (N) 20 h after stimulation of splenocytes ex vivo with DNA-LNP, plasmid DNA, or DNA-LNP in the presence of chemical inhibitors to the indicated DNA sensors. (O) Schematic of relevant pathways implicated in DNA-LNP sensing. Dots represent individual animals; n = 8–9 (E–H), n = 5 (I, K, and L), or n = 3–4 animals per group (L and M); data pooled or representative from two independent experiments (E–H, M, and N) or from one independent experiment (I–L). Plots show mean with SD (B–D and I) or geometric mean with geometric SD (F–H and K–N). Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (F–H, K, and L) or compared to DNA-LNP control (M and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Techniques Used: Zeta Potential Analyzer, Fluorescence, FACS, Enzyme-linked Immunospot, Luminex, Ex Vivo, Plasmid Preparation, Control

    HA DNA-LNP induces robust GC and serum responses Mice were immunized with HA DNA-LNP (2 μg), HA mRNA-LNP (2 μg), or adjuvanted HA protein (1 μg). GC responses were assessed in the DLNs 14 days post immunization and serum responses longitudinally. (A) Representative FACS plots of activated Tfh cells. (B and C) Bar plots show quantification of frequency (B) and numbers (C) of activated Tfh cells. (D) Representative FACS plots of total GC B cells. (E and F) Bar plots show quantification of frequency (E) and numbers (F) of total GC B cells. (G) Representative FACS plots of CA09 HA-specific GC B cells. (H and I) Bar plots show frequency (H) and numbers (I) of CA09 HA-specific GC B cells. (J) Area under the curve (AUC) of total A/California/04/2009 HA-specific serum IgG ELISA data. (K) Serum endpoint titers at week 8 to various H1N1 HAs. (L) HAI titers at week 8 to A/California/07/2009 X-179A. (M and N) AUC of serum binding antibodies to A/Guangdong-Maonan/SWL1536/2019 HA (M) and A/Victoria/4897/2022 HA (N). (O and P) HAI titers to A/Netherlands/602/2009 (O) and A/New York City/PV63249/2022 (P). Dots represent individual animals (B, C, E, F, H, I, K, and L); n = 9–10 animals per group; data pooled from two independent experiments. Plots show geometric mean with geometric SD. Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (A–I) or active immunization groups (K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
    Figure Legend Snippet: HA DNA-LNP induces robust GC and serum responses Mice were immunized with HA DNA-LNP (2 μg), HA mRNA-LNP (2 μg), or adjuvanted HA protein (1 μg). GC responses were assessed in the DLNs 14 days post immunization and serum responses longitudinally. (A) Representative FACS plots of activated Tfh cells. (B and C) Bar plots show quantification of frequency (B) and numbers (C) of activated Tfh cells. (D) Representative FACS plots of total GC B cells. (E and F) Bar plots show quantification of frequency (E) and numbers (F) of total GC B cells. (G) Representative FACS plots of CA09 HA-specific GC B cells. (H and I) Bar plots show frequency (H) and numbers (I) of CA09 HA-specific GC B cells. (J) Area under the curve (AUC) of total A/California/04/2009 HA-specific serum IgG ELISA data. (K) Serum endpoint titers at week 8 to various H1N1 HAs. (L) HAI titers at week 8 to A/California/07/2009 X-179A. (M and N) AUC of serum binding antibodies to A/Guangdong-Maonan/SWL1536/2019 HA (M) and A/Victoria/4897/2022 HA (N). (O and P) HAI titers to A/Netherlands/602/2009 (O) and A/New York City/PV63249/2022 (P). Dots represent individual animals (B, C, E, F, H, I, K, and L); n = 9–10 animals per group; data pooled from two independent experiments. Plots show geometric mean with geometric SD. Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (A–I) or active immunization groups (K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Binding Assay

    HA DNA-LNP induces potent memory responses in mice and rabbits (A) Schematic of mouse immunization regimen. (B) IFNγ-secreting cells in splenocytes by ELISpot. (C) IFNγ-secreting effector CD8 + T cells by flow cytometry. (D) CA09 HA-specific ASC responses in bone marrow by ELISpot. (E) Representative FACS plot of CA09 HA-specific MBCs. (F and G) Bar plots show frequency (F) and numbers (G) of CA09 HA-specific MBCs. (H) Schematic of rabbit immunization regimen. (I–K) IFNγ ELISpot on peripheral blood mononuclear cells (PBMCs) at day 42 (I), day 105 (J), and day 202 (K). (L) AUC of total A/California/04/2009 HA-specific serum IgG ELISA data. (M and N) HAI titers to A/Netherlands/602/2009 (M) and A/New York City/PV63249/2022 (N). Dots represent individual animals (C, D, F, and G); n = 9–10 animals per group (B–D, F, and G), n = 5 animals per group (I–N); data pooled from two independent experiments. Plots show mean with SD (B and I–K) or geometric mean with geometric SD (C, D, F, G, and L–N). Unpaired one-way or two-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups. ANOVA was performed at the final time point for (L–N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
    Figure Legend Snippet: HA DNA-LNP induces potent memory responses in mice and rabbits (A) Schematic of mouse immunization regimen. (B) IFNγ-secreting cells in splenocytes by ELISpot. (C) IFNγ-secreting effector CD8 + T cells by flow cytometry. (D) CA09 HA-specific ASC responses in bone marrow by ELISpot. (E) Representative FACS plot of CA09 HA-specific MBCs. (F and G) Bar plots show frequency (F) and numbers (G) of CA09 HA-specific MBCs. (H) Schematic of rabbit immunization regimen. (I–K) IFNγ ELISpot on peripheral blood mononuclear cells (PBMCs) at day 42 (I), day 105 (J), and day 202 (K). (L) AUC of total A/California/04/2009 HA-specific serum IgG ELISA data. (M and N) HAI titers to A/Netherlands/602/2009 (M) and A/New York City/PV63249/2022 (N). Dots represent individual animals (C, D, F, and G); n = 9–10 animals per group (B–D, F, and G), n = 5 animals per group (I–N); data pooled from two independent experiments. Plots show mean with SD (B and I–K) or geometric mean with geometric SD (C, D, F, G, and L–N). Unpaired one-way or two-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups. ANOVA was performed at the final time point for (L–N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Techniques Used: Enzyme-linked Immunospot, Flow Cytometry, Enzyme-linked Immunosorbent Assay


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Lysis, Reporter Gene Assay, Cell Stimulation, Electron Microscopy, Luminex, Enzyme-linked Immunospot, Luciferase, Plasmid Preparation, Software, Synthesized



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    Initial immune characterization of HA DNA-LNP formulations and immunogenicity (A) Schematic of DNA-LNP N/P ratios and immunization regimen. (B–D) Biophysical characterization of DNA-LNPs at different N/P ratios. (B) Particle size; (C) polydispersity index (PDI); (D) zeta potential. (E) Representative fluorescence-activated cell sorting (FACS) plots of GC B cells. (F) Bar plots quantifying frequency of GC B cells. (G) Frequency of CA09 HA-specific GC B cells. (H) Frequency of activated Tfh cells. (I) IFNγ ELISpot of splenocytes. (J) Representative TEM images of HA DNA-LNP (top) and HA mRNA-LNP (bottom). Scale bar 100 nm (K and L) Fold change cytokine induction in DLNs at 4 h (K) and 24 h (L) after immunization quantified using Luminex. (M and N) ELISpot assay measuring IFNα (M) and IFNγ (N) 20 h after stimulation of splenocytes ex vivo with DNA-LNP, plasmid DNA, or DNA-LNP in the presence of chemical inhibitors to the indicated DNA sensors. (O) Schematic of relevant pathways implicated in DNA-LNP sensing. Dots represent individual animals; n = 8–9 (E–H), n = 5 (I, K, and L), or n = 3–4 animals per group (L and M); data pooled or representative from two independent experiments (E–H, M, and N) or from one independent experiment (I–L). Plots show mean with SD (B–D and I) or geometric mean with geometric SD (F–H and K–N). Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (F–H, K, and L) or compared to DNA-LNP control (M and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Journal: Cell Reports Medicine

    Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

    doi: 10.1016/j.xcrm.2025.102035

    Figure Lengend Snippet: Initial immune characterization of HA DNA-LNP formulations and immunogenicity (A) Schematic of DNA-LNP N/P ratios and immunization regimen. (B–D) Biophysical characterization of DNA-LNPs at different N/P ratios. (B) Particle size; (C) polydispersity index (PDI); (D) zeta potential. (E) Representative fluorescence-activated cell sorting (FACS) plots of GC B cells. (F) Bar plots quantifying frequency of GC B cells. (G) Frequency of CA09 HA-specific GC B cells. (H) Frequency of activated Tfh cells. (I) IFNγ ELISpot of splenocytes. (J) Representative TEM images of HA DNA-LNP (top) and HA mRNA-LNP (bottom). Scale bar 100 nm (K and L) Fold change cytokine induction in DLNs at 4 h (K) and 24 h (L) after immunization quantified using Luminex. (M and N) ELISpot assay measuring IFNα (M) and IFNγ (N) 20 h after stimulation of splenocytes ex vivo with DNA-LNP, plasmid DNA, or DNA-LNP in the presence of chemical inhibitors to the indicated DNA sensors. (O) Schematic of relevant pathways implicated in DNA-LNP sensing. Dots represent individual animals; n = 8–9 (E–H), n = 5 (I, K, and L), or n = 3–4 animals per group (L and M); data pooled or representative from two independent experiments (E–H, M, and N) or from one independent experiment (I–L). Plots show mean with SD (B–D and I) or geometric mean with geometric SD (F–H and K–N). Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (F–H, K, and L) or compared to DNA-LNP control (M and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Article Snippet: For protein vaccination, 1 μg of recombinant full-length CA09 HA (Sino Biologicals Cat# 11055-VNAB) was formulated 1:1 v/v with Addavax Adjuvant (Cat# vax-adx-10, Invivogen).

    Techniques: Zeta Potential Analyzer, Fluorescence, FACS, Enzyme-linked Immunospot, Luminex, Ex Vivo, Plasmid Preparation, Control

    HA DNA-LNP induces robust GC and serum responses Mice were immunized with HA DNA-LNP (2 μg), HA mRNA-LNP (2 μg), or adjuvanted HA protein (1 μg). GC responses were assessed in the DLNs 14 days post immunization and serum responses longitudinally. (A) Representative FACS plots of activated Tfh cells. (B and C) Bar plots show quantification of frequency (B) and numbers (C) of activated Tfh cells. (D) Representative FACS plots of total GC B cells. (E and F) Bar plots show quantification of frequency (E) and numbers (F) of total GC B cells. (G) Representative FACS plots of CA09 HA-specific GC B cells. (H and I) Bar plots show frequency (H) and numbers (I) of CA09 HA-specific GC B cells. (J) Area under the curve (AUC) of total A/California/04/2009 HA-specific serum IgG ELISA data. (K) Serum endpoint titers at week 8 to various H1N1 HAs. (L) HAI titers at week 8 to A/California/07/2009 X-179A. (M and N) AUC of serum binding antibodies to A/Guangdong-Maonan/SWL1536/2019 HA (M) and A/Victoria/4897/2022 HA (N). (O and P) HAI titers to A/Netherlands/602/2009 (O) and A/New York City/PV63249/2022 (P). Dots represent individual animals (B, C, E, F, H, I, K, and L); n = 9–10 animals per group; data pooled from two independent experiments. Plots show geometric mean with geometric SD. Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (A–I) or active immunization groups (K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Journal: Cell Reports Medicine

    Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

    doi: 10.1016/j.xcrm.2025.102035

    Figure Lengend Snippet: HA DNA-LNP induces robust GC and serum responses Mice were immunized with HA DNA-LNP (2 μg), HA mRNA-LNP (2 μg), or adjuvanted HA protein (1 μg). GC responses were assessed in the DLNs 14 days post immunization and serum responses longitudinally. (A) Representative FACS plots of activated Tfh cells. (B and C) Bar plots show quantification of frequency (B) and numbers (C) of activated Tfh cells. (D) Representative FACS plots of total GC B cells. (E and F) Bar plots show quantification of frequency (E) and numbers (F) of total GC B cells. (G) Representative FACS plots of CA09 HA-specific GC B cells. (H and I) Bar plots show frequency (H) and numbers (I) of CA09 HA-specific GC B cells. (J) Area under the curve (AUC) of total A/California/04/2009 HA-specific serum IgG ELISA data. (K) Serum endpoint titers at week 8 to various H1N1 HAs. (L) HAI titers at week 8 to A/California/07/2009 X-179A. (M and N) AUC of serum binding antibodies to A/Guangdong-Maonan/SWL1536/2019 HA (M) and A/Victoria/4897/2022 HA (N). (O and P) HAI titers to A/Netherlands/602/2009 (O) and A/New York City/PV63249/2022 (P). Dots represent individual animals (B, C, E, F, H, I, K, and L); n = 9–10 animals per group; data pooled from two independent experiments. Plots show geometric mean with geometric SD. Unpaired one-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups (A–I) or active immunization groups (K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Article Snippet: For protein vaccination, 1 μg of recombinant full-length CA09 HA (Sino Biologicals Cat# 11055-VNAB) was formulated 1:1 v/v with Addavax Adjuvant (Cat# vax-adx-10, Invivogen).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

    HA DNA-LNP induces potent memory responses in mice and rabbits (A) Schematic of mouse immunization regimen. (B) IFNγ-secreting cells in splenocytes by ELISpot. (C) IFNγ-secreting effector CD8 + T cells by flow cytometry. (D) CA09 HA-specific ASC responses in bone marrow by ELISpot. (E) Representative FACS plot of CA09 HA-specific MBCs. (F and G) Bar plots show frequency (F) and numbers (G) of CA09 HA-specific MBCs. (H) Schematic of rabbit immunization regimen. (I–K) IFNγ ELISpot on peripheral blood mononuclear cells (PBMCs) at day 42 (I), day 105 (J), and day 202 (K). (L) AUC of total A/California/04/2009 HA-specific serum IgG ELISA data. (M and N) HAI titers to A/Netherlands/602/2009 (M) and A/New York City/PV63249/2022 (N). Dots represent individual animals (C, D, F, and G); n = 9–10 animals per group (B–D, F, and G), n = 5 animals per group (I–N); data pooled from two independent experiments. Plots show mean with SD (B and I–K) or geometric mean with geometric SD (C, D, F, G, and L–N). Unpaired one-way or two-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups. ANOVA was performed at the final time point for (L–N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Journal: Cell Reports Medicine

    Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

    doi: 10.1016/j.xcrm.2025.102035

    Figure Lengend Snippet: HA DNA-LNP induces potent memory responses in mice and rabbits (A) Schematic of mouse immunization regimen. (B) IFNγ-secreting cells in splenocytes by ELISpot. (C) IFNγ-secreting effector CD8 + T cells by flow cytometry. (D) CA09 HA-specific ASC responses in bone marrow by ELISpot. (E) Representative FACS plot of CA09 HA-specific MBCs. (F and G) Bar plots show frequency (F) and numbers (G) of CA09 HA-specific MBCs. (H) Schematic of rabbit immunization regimen. (I–K) IFNγ ELISpot on peripheral blood mononuclear cells (PBMCs) at day 42 (I), day 105 (J), and day 202 (K). (L) AUC of total A/California/04/2009 HA-specific serum IgG ELISA data. (M and N) HAI titers to A/Netherlands/602/2009 (M) and A/New York City/PV63249/2022 (N). Dots represent individual animals (C, D, F, and G); n = 9–10 animals per group (B–D, F, and G), n = 5 animals per group (I–N); data pooled from two independent experiments. Plots show mean with SD (B and I–K) or geometric mean with geometric SD (C, D, F, G, and L–N). Unpaired one-way or two-way ANOVA adjusted for multiple comparisons with Bonferroni corrections was used to compare groups. ANOVA was performed at the final time point for (L–N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

    Article Snippet: For protein vaccination, 1 μg of recombinant full-length CA09 HA (Sino Biologicals Cat# 11055-VNAB) was formulated 1:1 v/v with Addavax Adjuvant (Cat# vax-adx-10, Invivogen).

    Techniques: Enzyme-linked Immunospot, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    Journal: Cell Reports Medicine

    Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

    doi: 10.1016/j.xcrm.2025.102035

    Figure Lengend Snippet:

    Article Snippet: For protein vaccination, 1 μg of recombinant full-length CA09 HA (Sino Biologicals Cat# 11055-VNAB) was formulated 1:1 v/v with Addavax Adjuvant (Cat# vax-adx-10, Invivogen).

    Techniques: Virus, Recombinant, Lysis, Reporter Gene Assay, Cell Stimulation, Electron Microscopy, Luminex, Enzyme-linked Immunospot, Luciferase, Plasmid Preparation, Software, Synthesized

    (A) Dot plots representing expansion of Spike+ cells within total CD20+ B cells in PBMC before and after vaccination (aggregate differences on the left and matched differences on the right). PBMC were incubated with biotinylated Spike protein and fluorochrome conjugated streptavidin, surface stained, washed and analyzed using flow cytometry. Group differences were tested using unpaired t-test with Welch’s correction (left panel) or paired t-test (right panel). (B) Magnified view of B cell subsets identified using single cell RNA sequencing. Data includes samples from all four groups. (C) Waffle plot representation of B cell cluster quantification with infection and vaccination. (D) Isotype distribution of productive B cell clones in vaccinated (n=4) and convalescent (n=3) individuals. Isotypes were determined based on the constant region of the clone. (E) Aggregate clonal abundance following vaccination and infection. (F) Volcano plots depicting heavy chain gene usage biases following convalescence (left panel, relative to pre-infection baseline) or vaccination (bottom panel, relative to pre vaccination baseline). X axis represents the change in gene usage and Y axis represents p-value (−log 10). Two-way comparisons were tested using ratio-paired test for matched comparisons, [p-values: *− p<0.05]

    Journal: bioRxiv

    Article Title: Single cell profiling of T and B cell repertoires following SARS-CoV-2 mRNA vaccine

    doi: 10.1101/2021.07.14.452381

    Figure Lengend Snippet: (A) Dot plots representing expansion of Spike+ cells within total CD20+ B cells in PBMC before and after vaccination (aggregate differences on the left and matched differences on the right). PBMC were incubated with biotinylated Spike protein and fluorochrome conjugated streptavidin, surface stained, washed and analyzed using flow cytometry. Group differences were tested using unpaired t-test with Welch’s correction (left panel) or paired t-test (right panel). (B) Magnified view of B cell subsets identified using single cell RNA sequencing. Data includes samples from all four groups. (C) Waffle plot representation of B cell cluster quantification with infection and vaccination. (D) Isotype distribution of productive B cell clones in vaccinated (n=4) and convalescent (n=3) individuals. Isotypes were determined based on the constant region of the clone. (E) Aggregate clonal abundance following vaccination and infection. (F) Volcano plots depicting heavy chain gene usage biases following convalescence (left panel, relative to pre-infection baseline) or vaccination (bottom panel, relative to pre vaccination baseline). X axis represents the change in gene usage and Y axis represents p-value (−log 10). Two-way comparisons were tested using ratio-paired test for matched comparisons, [p-values: *− p<0.05]

    Article Snippet: To detect antigen specific B cells, ~ 5×10e5 PBMC were stained with 100 ng of full length biotinylated spike protein (Sino Biological) pre-incubated with Streptavidin-BV510 (Biolegend) at 2:1 ratio for 1 h at 4C to ensure maximum staining quality before surface staining with CD20-FITC (2H7, Biolegend) for an additional 30 minutes.

    Techniques: Incubation, Staining, Flow Cytometry, RNA Sequencing Assay, Infection, Clone Assay