Journal: Future Virology
Article Title: Design and generation of mRNAs encoding conserved regions of SARS-CoV-2 ORF1ab for T cell-mediated immune activation
doi: 10.2217/fvl-2023-0066
Figure Lengend Snippet: (A) Representative ELISpots of PBMCs transfected with S, P1, P2, no mRNA (mock) or PBMCs treated with ConA (positive control) from participant 6 (responder) and 26 (non-responder). The number of spot counts per well was indicated. (B) Fold increase of IFN-γ spot counts of PBMCs after transfection with S, P1, or P2 mRNA relative to the mock treatment from 34 participants. Participants having a fold increase less than 2 (cutoff value) were classified as non-responders. (C) UpSet plot showing the number of participants having T-cell responses to S, P1 or P2 mRNA, as well as the overlap between such conditions. Percentages of (D) CD4 + IFN-γ + T cells and (E) CD8 + IFN-γ + T cells measured by flowcytometry from 34 participants, grouped into <1 month, >1 month, and unknown, after transfection with S, P1, or P2 mRNA. One-way ANOVA test was performed to compare the frequencies of CD4 + IFN-γ + T cells and CD8 + IFN-γ + T cells across different groups; *p < 0.05; **p < 0.01; ***p < 0.001.
Article Snippet: To evaluate the T-cell response when stimulated with the mRNA, Human IFN-γ ELISpotPRO kit (ALP) (Mabtech, 3420-2AST-2, Nacka Strand, Sweden) was used to measure IFN-γ secretion by PBMCs, following the manufacturer's instruction.
Techniques: Transfection, Positive Control