A Novel Class of Small Molecule Agonists with Preference for Human over Mouse TLR4 Activation
Fig 9
Species-specific activation of IL-6, TNF-α, and chemokine expression in primary cells.
PBMCs were isolated from peripheral blood from human healthy volunteers, cynomolgus macaques, Hartley guinea pigs, ferrets, New Zealand white rabbits, Sprague-Dawley rats, and cotton rats, and splenocytes were isolated from BALB/c mice, n = 3 for each species, except n = 2 for cotton rats. TLR4 agonists were used at 1 μg/ml to stimulate primary cells of all species for 24 hrs. For human, cyno, mouse, and rat, SNs were analyzed via human, NHP, mouse, and rat MILLIPLEX kits. For guinea pig, ferret, rabbit, and cotton rat, cell lysates were analyzed via Affymetrix QuantiGene Plex kits. No one chemokine analyte was available in all 8 species-specific assays, so data were collected for any one of three different chemokines, MCP-1, MIP-1β, and RANTES. Data are expressed as mean compound % difference from the mean MPL value for that species and cytokine with associated 90% confidence interval (CI) listed below. A compound is considered significantly higher than MPL if the lower bound of the CI is higher than 0% (p value < 0.05); such values are in bold. Increasing shades of green and purple indicate increasingly higher and lower respective values than values achieved by MPL stimulation. Raw data are presented in S6 Fig.