chikv e2  (Sino Biological)

Journal: Signal Transduction and Targeted Therapy

Article Title: CHIKV mRNA vaccines encoding conserved structural/envelope proteins confer broad cross-lineage protection against infection

doi: 10.1038/s41392-025-02182-2

Figure Lengend Snippet: Vaccine design, preparation, and in vitro expression. a Vaccine design. A total of 769 strains were identified from the NCBI database, including 14 from the West African lineage, 335 from the Asian lineage, 121 from the East, Central, and South African lineage, and 299 from the Indian Ocean lineage. Conserved sequences were selected for further analysis; b Utilize Alpha-Fold3 for structural prediction and receptor interaction prediction of the E protein in vaccine sequences and viral sequences (MH670649.1); c Dynamic light scattering (DLS) particle size distribution; d Transmission electron microscopy (TEM) images; e E1, E2, C protein, and β-actin Western blot images; (−) indicates the negative control, and M indicates the control transfected with a commercial kit; f Immunofluorescence experiments were conducted to detect the distribution of the E1 protein, using nontransfected cells as the mock condition

Article Snippet: E2 protein (Sino Biological, #40440-V08B, 1 μg/ml, 100 μl/well) was coated in a 96-well plate (442404, Thermo Fisher Scientific) overnight at 4 °C.

Techniques: In Vitro, Expressing, Transmission Assay, Electron Microscopy, Western Blot, Negative Control, Control, Transfection, Immunofluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: CHIKV mRNA vaccines encoding conserved structural/envelope proteins confer broad cross-lineage protection against infection

doi: 10.1038/s41392-025-02182-2

Figure Lengend Snippet: Cellular immune response assessment. a – f Elispot experiments to enumerate specific spots indicating cytokine secretion by splenocytes stimulated with the E2 protein for IFN-γ ( a ), IL-2 ( c ), and IL-4 ( e ) and stimulated with the inactivated virus for IFN-γ ( b ), IL-2 ( d ), and IL-4 ( f ); g – j Flow cytometry analysis of the percentages of CD4+ T cells producing IL-2 ( g ), IL-4 ( h ), IFN-γ ( i ), and TNF-α ( j ); k – m The proportions of CD8+ T cells producing IL-2 ( k ), IFN-γ ( l ), and TNF-α ( m ). The data are presented as the mean ± SEM ( n = 5 or 3). Statistical analysis was conducted using two-way ANOVA and Tukey’s multiple comparison test for bar graphs; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns not significant

Article Snippet: E2 protein (Sino Biological, #40440-V08B, 1 μg/ml, 100 μl/well) was coated in a 96-well plate (442404, Thermo Fisher Scientific) overnight at 4 °C.

Techniques: Enzyme-linked Immunospot, Virus, Flow Cytometry, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: CHIKV mRNA vaccines encoding conserved structural/envelope proteins confer broad cross-lineage protection against infection

doi: 10.1038/s41392-025-02182-2

Figure Lengend Snippet: mCV-1 and mCV-2 protect A129 mice from mortality due to CHIKV infection. a The vaccination and sample collection timeline, Serum collection, and determination of binding antibody titers and neutralizing antibody levels were performed on days 21 and 28 after initial immunization, followed by challenge experiments on day 42. Body temperature, body weight, and joint changes were monitored daily after the challenge until all mice in the placebo group died, and survival curves were plotted; b The titers of binding antibodies at days 21 and 28; c Neutralizing antibody titers at 21, 28, and 56 days post-immunization; d Variation in viremia within 21 days post-challenge; e and f Changes in body weight ( e ) and the degree of swelling in the right hind limb joints ( f ) of A129 mice post-challenge; g Survival rate of mice post-challenge; h Viral loads in the spleen and joint tissues of mice on day 21 post-challenge; i Elispots experiments to enumerate specific spots indicating cytokine secretion by splenocytes stimulated with the E2 protein for IFN-γ, IL-2, and IL-4; j-k, Joint tissue pathological sections ( k ) and pathological scores ( j ). Statistical analysis was conducted using one-way ANOVA and Tukey’s multiple comparison tests for bar graphs; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns not significant

Article Snippet: E2 protein (Sino Biological, #40440-V08B, 1 μg/ml, 100 μl/well) was coated in a 96-well plate (442404, Thermo Fisher Scientific) overnight at 4 °C.

Techniques: Infection, Binding Assay, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: CHIKV mRNA vaccines encoding conserved structural/envelope proteins confer broad cross-lineage protection against infection

doi: 10.1038/s41392-025-02182-2

Figure Lengend Snippet: mCV-1 and mCV-2 protect rhesus macaques from CHIKV challenge. a The immunization schedule for rhesus macaques is depicted in the flowchart, with vaccinations administered on Day 0 and Day 21, at a dosage of 300 μg per animal, using physiological saline as the placebo group. Serum samples were collected on Days 7, 14, 21, 28, and 35 post-primary immunization, followed by a challenging experiment on Day 42 ( n = 3); b and c The titers of binding antibodies ( b ) and neutralizing antibodies against adapted strains ( c ) at different time points; d GMTs of pseudovirus neutralizing antibodies against various strains at 35-day post-immunization for mCV-1 and mCV-2; e Elispot experiments to enumerate specific spots indicating cytokine secretion by splenocytes stimulated with the E2 protein for IFN-γ, IL-2, and IL-4; f Changes in viremia within 7 days post-immunization with mCV-1 and mCV-2; g Tissue viral loads in rhesus macaques immunized with mCV-1, mCV-2 or physiological saline on day 7 post-challenge, LN: Hilar lymph nodes, Kid: Kidney, PA: Pancreas, ILN: Inguinal lymph nodes, MLN: mesenteric lymph nodes, SMG: submandibular lymph node; h and i pathological scores ( h ) and pathological sections ( i ). The data are presented as The mean ± SEM ( n = 3), and each symbol represents a mouse. Statistical analysis was conducted using one-way ANOVA and Tukey’s multiple comparison tests for bar graphs; *** p < 0.001; **** p < 0.0001; ns not significant

Article Snippet: E2 protein (Sino Biological, #40440-V08B, 1 μg/ml, 100 μl/well) was coated in a 96-well plate (442404, Thermo Fisher Scientific) overnight at 4 °C.

Techniques: Saline, Binding Assay, Enzyme-linked Immunospot, Comparison