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: a Schematic view of chemical structure of SPA14 liposomes formed by DOPC, cholesterol, QS21 and E6020 consisting of a hexa-acylated acyclic structure backbone. b Solubility of E6020 and MPLA in ethanol. E6020 and MPLA (Sigma) powders were suspended at 0.5, 1.0, 2.0 and 10 mg/mL in absolute ethanol and solubilization was followed by nephelometry as described in the Methods section. The RNU (Relative Nephelometry Unit) of each sample was recorded and plotted on the graph. c Cryo EM micrograph for SPA14-20 (scale = 100 nm) with unilamellar and few multilamellar vesicles. d Hemolytic activity of QS21 and quenching of hemolysis in SPA14 and AS01B. The graph shows hemolysis of sheep erythrocytes incubated with increasing concentrations of QS21 either in free form (solution in CBS pH 6.3; green line) or formulated in SPA14-8 (blue line) or in AS01B (red line).

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: Liposomes, Solubility, Cryo-EM Sample Prep, Activity Assay, Incubation

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: Adjuvants evaluated in the MIMIC® PTE module

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: Concentration Assay

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

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: Data for SPA14-20, SPA14-8, and liposome at 1:40 dilution and AS01B at the 1:20 dilution are represented here as to normalize DOPC-Chol-QS21 contents in the different formulations. E6020 (200 ng/mL, corresponding to a 1:40 dilution of SPA14) MPLA (5000 ng/mL, corresponding to a 1:20 dilution of AS01B), and liposome (SPA14 without E6020 at a 1:40 dilution) were evaluated along with a LPS + R848 (L + R) positive control. Cytokines and chemokines were organized based on their alignment with the TLR4- intracellular signaling pathways, MyD88 and TRIF. N = 8–15 donors.

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: Positive Control