full length ca09 ha  (Sino Biological)

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

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

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

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: The construct encoding the full-length SARS-CoV-2 spike glycoprotein (with the D614G mutation, which arose early during the COVID-19 pandemic) has been described previously., , Full length CA09 HA was cloned into a pVAX1 vector behind an IgE leader sequence and obtained from Genscript. mRNA vaccines encoding the full-length wild-type SARS-CoV-2 spike glycoprotein were synthesized as described in Laczkó et al. CA09 HA mRNA-LNPs were described previously.

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

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: The construct encoding the full-length SARS-CoV-2 spike glycoprotein (with the D614G mutation, which arose early during the COVID-19 pandemic) has been described previously., , Full length CA09 HA was cloned into a pVAX1 vector behind an IgE leader sequence and obtained from Genscript. mRNA vaccines encoding the full-length wild-type SARS-CoV-2 spike glycoprotein were synthesized as described in Laczkó et al. CA09 HA mRNA-LNPs were described previously.

Techniques: Enzyme-linked Immunosorbent Assay, Binding 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: 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: The construct encoding the full-length SARS-CoV-2 spike glycoprotein (with the D614G mutation, which arose early during the COVID-19 pandemic) has been described previously., , Full length CA09 HA was cloned into a pVAX1 vector behind an IgE leader sequence and obtained from Genscript. mRNA vaccines encoding the full-length wild-type SARS-CoV-2 spike glycoprotein were synthesized as described in Laczkó et al. CA09 HA mRNA-LNPs were described previously.

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: The construct encoding the full-length SARS-CoV-2 spike glycoprotein (with the D614G mutation, which arose early during the COVID-19 pandemic) has been described previously., , Full length CA09 HA was cloned into a pVAX1 vector behind an IgE leader sequence and obtained from Genscript. mRNA vaccines encoding the full-length wild-type SARS-CoV-2 spike glycoprotein were synthesized as described in Laczkó et al. CA09 HA mRNA-LNPs were described previously.

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