Using qRT-PCR, we found that WT MΦ treated with TNFα 30 min prior to infection with O. tsutsugamushi (5 MOI) exhibited significantly greater Mincle expression (25-fold increase, p < 0.0001) at 4 hr after infection than cells receiving TNFα (20-fold increase, p < 0.001) or O. tsutsugamushi alone (8-fold increase, p < 0.01, Fig 9). This evidence concerns the gene CLEC4E and infection.