Fig. 1

Monocyte response to TLR7/TLR8 stimulation. A Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of pregnant (n = 20) and non-pregnant (n = 20) women and stimulated with R848 (TLR7/TLR8 agonist) or Gardiquimod (TLR7 agonist) for 24 h. Flow cytometry was performed to phenotype monocytes. B Flow cytometry gating strategy for phenotyping of monocyte subsets after in vitro stimulation with viral ligands. Viable monocytes were gated as live CD14⁺ cells from PBMCs. The expression levels of CD16 and CD14 were used to gate monocyte subsets as follows: classical (CD14^hiCD16⁻); intermediate (CD14^hiCD16⁺); non-classical (CD14^loCD16⁺), and CD14^loCD16⁻. C Proportions of monocyte subsets in pregnant (red) and non-pregnant (blue) women with and without R848 stimulation. D Heatmap representation of the differences in proportions of monocytes subsets from pregnant (red symbols) and non-pregnant (blue symbols) following R848 stimulation. Asterisks indicate statistically significant differences between the indicated groups. E-N Frequencies of (E) classical monocytes, (F) intermediate monocytes, (G) non-classical monocytes, (H) CD14^loCD16⁻ monocytes, (I) CD147⁺ monocytes, (J) CD162⁺ monocytes, (K) CCR5⁺CCR2⁺ monocytes, (L) CCR5⁻CCR2⁺ monocytes, (M) CCR5⁻CCR2⁻ monocytes, and (N) CCR5⁺CCR2⁻ monocytes in pregnant (red) and non-pregnant (blue) women following R848 stimulation (solid circles) or control (open circles). *p < 0.05; **p < 0.01; ***p < 0.001. ( +) Stimulated; (-) Control