tmprss2 expression constructs  (Addgene inc)

Journal: iScience

Article Title: Flavivirus-based bivalent nanoparticle vaccines induce neutralizing antibodies and Th1 responses against flavivirus and coupling antigens

doi: 10.1016/j.isci.2025.113659

Figure Lengend Snippet: Induction of antigen-specific neutralizing antibodies against both JEV and SARS-CoV-2 by immunization with JEprME-S-RBD nanoparticles in mice (A) BALB/c mice ( n = 4–6 per group) were injected in the quadriceps muscle with vehicle (group 1), 10 μg of soluble S-RBD (roup 2), 1 μg of JEprME-S-RBD particles containing S-RBD (group 3), and non-binding mixture (JEprME-SpyTag and soluble S-RBD) (group 4) with alum adjuvant at 2-week intervals. In all groups, the inoculum was given in alum adjuvant, an aluminum hydroxide gel suspension. We harvested the quadriceps muscle and sera 2 weeks after the second immunization. Results are representative of two experiments. (B) Histology of the injected quadriceps muscle of each group. We visualized the inoculum remaining in the muscle by aluminum-specific lumogallion staining, and inflammatory cells by hematoxylin and eosin (H&E) staining. (C and D) Comparison of the levels of inflammation and remaining inoculum among groups. Each closed circle represents an individual mouse, and the bars indicate the mean values for each group. Lumogallion staining was negative in the PBS group; aluminum was not detected (N.D). ∗ p < 0.05. We semi-quantified the levels of inflammation and remaining inoculum in the muscle. The inflammatory or lumogallion-positive areas were divided by the whole muscle areas to obtain the percentage of inflammatory or aluminum-containing areas. (E) Changes in body weight of immunized mice. The y axis represents the body weight (g) of mice, and the x axis represents the days after the first immunization. (F and G) Levels of S-RBD-specific (F) and JE-prME-specific (G) IgG in sera. Sera were diluted 2-fold from a 100-fold dilution, and IgG levels against each antigen were detected by enzyme-linked immunosorbent assays (ELISA). Thin lines represent the IgG levels of individual mice; open circles and bold lines represent the mean IgG values of each group. Error bars represent standard deviation ( n = 5). (H) Serum neutralizing activities against SARS-CoV-2 Wuhan (D614G) strain pseudovirus. Serially 2-fold diluted sera were reacted with the pseudovirus, followed by infection of VeroE6-TMPRSS2 cells and measurement of NanoLuc luciferase activity in cell lysates 48 h later. (I) Serum neutralizing activities against JEV. Serially 2-fold diluted antisera were reacted with the JEV-NS1-HiBiT reporter virus, followed by infection of Vero cells and measurement of HiBiT activity in culture supernatants 72 h later.

Article Snippet: Plasmid: TMPRSS2 , Edie et al. , Addgene Plasmid: #53887.

Techniques: Injection, Binding Assay, Adjuvant, Suspension, Staining, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation, Infection, Luciferase, Activity Assay, Virus

Journal: iScience

Article Title: Flavivirus-based bivalent nanoparticle vaccines induce neutralizing antibodies and Th1 responses against flavivirus and coupling antigens

doi: 10.1016/j.isci.2025.113659

Figure Lengend Snippet: Serum neutralization activities against five SARS-CoV-2 variants Serum neutralization assays were conducted against pseudoviruses of the Alpha variant (A), Beta variant (B), Delta variant (C), Omicron BA.1 variant (D), and Omicron BA.2.75 variant (E). After reacting each pseudovirus with sera from each group, VeroE6-TMPRSS2 cells were infected, and luciferase activities were measured in cell lysates 48 h post-infection. Sera were diluted 2-fold from the original concentration. Each circle represents one mouse ( n = 5), and bars indicate the mean values for each group. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗, p < 0.0001.

Article Snippet: Plasmid: TMPRSS2 , Edie et al. , Addgene Plasmid: #53887.

Techniques: Neutralization, Variant Assay, Infection, Luciferase, Concentration Assay