Fig 1.
Pregnancy does not affect maternal parasite burden and Ld-induced anemia and splenomegaly.
(A) Peripheral blood hematocrit, RBC count, and WBC count of naïve and Ld-infected dams on the 18th day of pregnancy (n = 8-21). Pregnancy does not worsen Ld-induced anemia. (B) Spleen and liver weights of naïve and Ld-infected dams on the 18th day of pregnancy (n = 8-21). Pregnancy does not worsen Ld-induced splenomegaly. (C) Leishman-Donovan Units (LDU) in the spleen and liver of naïve and Ld-infected dams on the 18th day of pregnancy (n = 10-21). Pregnancy does not affect parasite burden in the spleen and liver of Ld-infected dams. Open circles indicate the naïve group, and closed circles indicate the Ld-infected group. Means ± SE are presented. P values for Student’s t test or two-way ANOVA with Sidak multiple comparisons are shown.
Fig 2.
Pregnancy-induced decrease in maternal serum BUN and IFN-
γ. (A) Serum BUN, CRE, AST, and ALT levels in dams on day 18 of pregnancy (n = 8-21). Pregnancy reduced Ld-induced serum BUN elevation. (B) Serum IFN-γ, IL-4, and TNF-α levels in dams on day 18 of pregnancy (n = 5-14). Pregnancy reduced Ld-induced serum IFN-γ elevation. Open circles indicate the naïve group, and closed circles indicate the Ld-infected group. Means ± SE are presented. P values for two-way ANOVA with Sidak multiple comparisons are shown. n.d., not detected.
Table 1.
The pregnancy rate.
Fig 3.
Reduced mating success rate due to Ld infection.
(A) Changes in the negative rate of vaginal plugs after the start of mating (n = 17-21). Ld-infected mice require a longer mating period than naive controls to detect vaginal plug. (B) Left: Change in maternal body weight after pregnancy (n = 16-21). Right: Weight gain rate of the dams on day 18 of pregnancy. Maternal body weight changes after pregnancy are not affected by Ld infection. Individuals who miscarried during the process are excluded. (C) Number of implants, resorption rate, number of fetuses, fetal weight, and placental weight on the 18th day of pregnancy (n = 16-21). For fetal weight and placental weight, the average of each dam’s offspring is plotted. None of the parameters were affected by Ld infection. (D) Left: Relationship between fetal number and maternal spleen/liver weight (n = 21). Right: Relationship between fetal number and maternal spleen/ liver LDU (n = 21). No correlation was observed between fetal number and maternal VL severity. Means ± SE are presented. P values for Log-rank test, Student’s t test or two-way ANOVA with Sidak multiple comparisons are shown. n.s., not significant (p > 0.05).
Table 2.
PCR positive rates of offspring.
Fig 4.
Vertical transmission of Ld with placental degeneration
(A) Estimated Ld number in the spleens and livers of vertically transmitted fetuses (n = 58-65). Means ± SE are presented. P value for Mann-Whitney test is shown. (B) Detection of KMP11-positive cells in fetal liver. (C) Left: Ld-induced placental degeneration. Trophoblastic cell atrophy (arrow) and vasodilation (arrowhead) were observed in HE-stained infected placentas. Bars, 20 μm. Right: Labyrinth zone tissue area excluding vascular lumen at 200 × magnification (n = 5-21). Means ± SE are presented. P value for Student’s t test is shown.
Fig 5.
Villitis with T cell infiltration in the placenta of Ld-infected mice.
(A) The expression of CD206, MRP14, B220 and CD3 in the placentas. Ld infection decreased CD206+ MΦs and MRP14+ MΦs, and increased CD3+ T cells in the placentas. (B) Consistent localization of CD3, ICAM1, and VCAM1 in serial placental sections. (C) cNK and uNK cells in DBA/PAS-stained placental sections. Open arrowhead indicates cNK cells (DBA- PAS+), and closed arrowhead indicates uNK cells (DBA+ PAS+). The number of cells was counted in 5 random microscopic fields of placental labyrinth zone at 400 × magnification (n = 5-21). Bars, 50 μm. Means ± SE are presented. P values for Student’s t test are shown.
Table 3.
RNA-Seq transcriptome analysis of immune factors in placentas of pups born to Ld-infected dams.