Journal: NPJ Vaccines
Article Title: SPA14 liposomes combining saponin with fully synthetic TLR4 agonist provide adjuvanticity to hCMV vaccine candidate
doi: 10.1038/s41541-024-01046-0
Figure Lengend Snippet: The MIMIC® PTE was treated with various human dose dilutions of adjuvants for 48 h, as described in Table . The cells were then harvested and evaluated for phenotype and viability by flow cytometry and cytokines by multiplex array. a Cell viability: % of viable cells after 48 h in MIMIC PTE assay. b APC differentiation was measured by the number of mature CD86+ cells as a percent of parent CD11c+ HLA DR+ cells. c IL-6 secretion as a representative cytokine determined in the culture supernatants by Luminex-based multiplex assay. d PGE-2 secretion determined by ELISA in the culture supernatants. e the level of surface markers (CD14, CD86 and CD83) on APCs (CD11c HLA + DR+ cells) measured by flow cytometry using specific antibodies. Data for SPA14-20, SPA14-8 and liposomes (SPA14 without E6020) at a 1:40 dilution and data for AS01B at a 1:20 dilution to normalize DOPC-Chol-QS21 contents between the 3 adjuvants are represented here. E6020 (200 ng/mL corresponding to a 1:40 dilution of SPA14), MPLA (5000 ng/mL corresponding to a 1:20 dilution of AS01B), liposomes and the L + R assay positive control were used as benchmarks in the study (see Methods). Each dot represents an individual donor and the bar represents Geometric Mean with 95% CI; N = 8–15 donors. Statistical comparisons were performed using GraphPad Prism by unpaired, two tailed T test for relevant conditions.
Article Snippet: For this purpose we developed a saponin-based adjuvant system by combining QS21 with liposomes comprising the well-defined fully synthetic clinical stage TLR4 agonist developed by Eisai Inc. and identified as E6020 .
Techniques: Flow Cytometry, Multiplex Assay, Luminex, Enzyme-linked Immunosorbent Assay, Liposomes, Positive Control, Two Tailed Test