Complete deletion of the Chlamydia muridarum putative cytotoxin locus reveals contributions during invasion in tissue culture and oviduct pathology during murine genital tract infection (Mouse data)

Figure 5. C57Bl/6 female mice were infected intravaginally with 105 IFU of C. muridarum WT, Δtox, or toxRep. A) IFU enumerated from vaginal swabs collected at days 3-28 post infection. Data presented as means +/- standard deviation from five mice/group/time point. Statistical significance was assessed by two-way RM ANOVA followed by Tukey’s multiple comparisons test; p < 0.05 for WT vs. other strains at day 3 and 7. B) Ascension at day 7 post intravaginal infection as measured by qPCR on upper genital tract homogenates. Statistical significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons test and found to be not significant. C) Frequency of hydrosalpinx at 51 days post intravaginal infection, pooled from two independent experiments..Figure 6. C57Bl/6 female mice were infected transcervically with 106 IFU of C. muridarum WT, Δtox or toxRep. At three days (A) and seven days (B) post-infection, bacterial burden in the upper genital tract was quantified by qPCR. Data are plotted as mean +/- SEM. Data in B are pooled from two independent experiments. Statistical significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons test; ns: not significant. Figure 7. Seven days after transcervical infection of C57Bl/6 mice with 106 IFU of C. muridarum WT, Δtox or toxRep, female genital tracts were excised and histopathology was assessed on randomized, anonymized slides. A-C) Pathology scores and representative images of H&E stained oviduct (OD, A), ovary (OV, B), and uterus (UT, C). Figure 8. Fifty-one days after transcervical infection of C57Bl/6 mice with 106 IFU of C. muridarum WT, Δtox or toxRep, female genital tracts were assessed for gross pathology. A) Representative images of excised upper genital tracts; asterisk indicates hydrosalpinx. B) Frequency of hydrosalpinx pooled from two independent experiments. Statistical significance was assessed by Fisher’s exact test; **, P = 0.0078, *, P = 0.0209, ns: not significant. Figure 9. Fifty-one days after transcervical infection of C57Bl/6 mice with 106 IFU of C. muridarum WT, Δtox or toxRep, female genital tracts were excised and histopathology was assessed on randomized, anonymized slides. A) Oviduct (OD) pathology scores. B) Representative images of H&E-stained oviduct and ovary. C) Ovary (OV) pathology scores. D) Pathology scores and representative images of H&E-stained uterus (UT). Two images of uterus are shown from each infection condition to represent the within-group variability of horn dilation that was present after infection with all three bacterial strains. Table S2. WT, tox and cis-tox were subjected to whole genome sequencing and variant analysis was done using C. muridarum Nigg AE002160.2