Figure S1.
B. abortus infection can be transferred by BM transplantation, and all B. abortus strains are equally virulent. (a) Experimental scheme: Mice were intraperitoneally inoculated with 1 × 106 CFU of wild-type B. abortus. BM cells were transplanted into previously lethally irradiated mice. 8 wk after transplantation, CFU per gram of organ was enumerated from spleens and BM. (b) Enumeration of CFU per gram of spleen and BM 8 wk after transplantation (n = 7). (c and d) wt and CD150−/− mice were intraperitoneally injected with PBS or inoculated with 1 × 106 CFU of B. abortus. 8 d later, BM cells were isolated, cells were counted (c), and then depleted for mature hematopoietic cells as shown in the Materials and methods. Lin− cells (d) were also counted for mice injected with PBS (Mock, unfilled circle) or infected B. abortus (Ba WT; black square), B. abortus ∆omp25 (Ba ∆omp25; gray square) or B. abortus ∆omp25 complemented with p:Omp25 (Ba ∆omp25c; unfilled square) mutants (the latter only for wt mice). From left to right: for BM, n = 11, 14, 8, 5, 9, 11, 9; and for lin− BM, n = 18, 16, 8, 6, 6, 7, 9. Data were obtained from distinct samples from five independent experiments. (e and f) CFU counts per gram of organ at day 2 (e) and day 8 (f) after infection for spleen and BM of mice infected with Ba WT (black square), Ba ∆omp25 (gray square) or Ba ∆omp25c (unfilled square). For day 2 (D2) spleen, n = 9, 6, 8, 8; for day 2 (D2) BM, n = 6, 6, 7, 7; for day 8 (D8) spleen, n = 19, 15, 9, 15, 23, 6; for day 8 (D8) BM, n = 5, 11, 4. Data were obtained from distinct samples from four independent experiments (a–d), each with at least three animals. Mean ± SEM is represented by a horizontal bar. Significant differences from mock are shown. Absence of P value or ns, non-significant. Since data did not follow normal distribution, P values were generated using Kruskal–Wallis followed by Dunn’s test. (g) Contribution of HSC from wt CD45.1 (left panel) and CD150−/− CD45.1 (right panel) mice incubated for 30 min ex vivo with Ba WT (black square) or Ba ∆omp25 (gray square) or non-infected (Mock, unfilled circle) as described in Fig. 3 e, to blood chimerism in CD45.2 recipients, at 4, 6, and 8 wk after transplantation (from left to right: for WT, n = 12, 13, 10, 10, 9, 9, 12, 8, 8; and for CD150−/−, n = 9, 4, 14, 8, 11, 7). Data were obtained from repetitive sampling from two independent experiments. Mean ± SEM is represented by horizontal bar. Absence of P value, non-significant. Since data did not follow normal distribution, P values were generated using Kruskal–Wallis followed by Dunn’s test.