IL-10 levels in infected pregnant B6 mice were statistically significantly reduced relative to infected pregnant A/J mice GSK3235025 on experiment day 9 (median (IQR): 36 (0–46) pg/mL for B6 vs. 550 (431–735) pg/mL

for A/J; P = 0·001), but this pattern was reversed on experiment day 10 (Figure 4a). In both strains, IL-10 levels were enhanced at experiment day 11 in infected pregnant relative to uninfected pregnant mice. Levels of sTNFRII did not differ between infected pregnant A/J and B6 mice at any of the tested time points, although the levels were consistently statistically significantly higher in the infected mice relative to their within strain uninfected counterparts (Figure 4c, d and data not shown). At none of the time points were the differences in IL-10 or sTNFRII observed between infected pregnant and infected non-pregnant mice of either strain nor were across-strain differences between infected non-pregnant mice found (Figure 4). To further evaluate immune changes associated with P. chabaudi AS infection and pregnancy loss in A/J and B6 mice, phenotypes RNA Synthesis inhibitor and levels of splenic leucocyte subsets were analysed flow cytometrically at experiment days 9 and 10, time points at which mice of both strains retain a proportion of viable conceptuses. No statistically significant differences in B-, natural killer (NK) or T-cell counts, including T-cell subsets, were observed between infected pregnant

A/J and B6 mice (Figure 5). However, malarial infection clearly stimulated expansion of all of these cell types in pregnant A/J mice, in whom splenocyte numbers for all subsets (except T cells at experiment day 9) were statistically significantly higher relative to their uninfected pregnant counterparts (Figure 5).

Similar differences in B6 mice were noted only for T, CD8+ T, B and NK cells on experiment day 9, but not on 10 (Figure 5). The total number of lymphocytes and lymphocyte subsets in general did not differ between infected pregnant and infected non-pregnant mice within each strain; only CD4+ T cells on experiment day 9 were significantly expanded in infected pregnant Dapagliflozin relative to infected non-pregnant A/J mice (Figure 5c). Similar to the lymphocyte subsets, numbers of neutrophils, monocytes and monocytes with an inflammatory phenotype (CD11b+/CD115+/Gr1high) were similar in the infected pregnant B6 and A/J mouse spleens on experiment days 9 and 10 (Figure 6). Neutrophil levels were enhanced in infected B6 mice at experiment day 9 relative to uninfected pregnant B6 mice (Figure 6a), a difference that did not reach statistical significance on experiment day 10 (Figure 6b: Kruskal–Wallis, P = 0·0024; Dunn’s pairwise comparisons, all P > 0·05). Monocytes levels were increased in infected pregnant B6 mice compared to their uninfected counterparts on experiment days 9 and 10, and on the former day were also higher than in infected non-pregnant mice (Figure 6c, d).