Fig 1.
Phagocytic activity of BALB/c and XID APerC after 1 h of infection with E. cuniculi spores by ultramicrography.
(A and insert) Projections of the cellular membrane (head arrows) of macrophages near or involving the spores (arrow) of E. cuniculi in XID APerC. (B and insert) Projections of the cellular membrane of macrophages involving E. cuniculi spore (arrow) of in BALB/c APerC and amorphous material (*) inside phagosome vacuole. Note the presence of mast cell. (C) Homotypic phagosomes vacuoles with E. cuniculi spores inside (arrow) and amorphous material exocytosed (head arrow) in BALB/c APerC macrophages. (D) Homotypic phagosomes vacuoles with intact spores (arrow) and megasome (me) with amorphous material inside in the macrophages (M) from XID APerC. Note mast cells (Mc) in contact with macrophages.
Fig 2.
Phagocytic activity of BALB/c and XID APerC after 48 hours of infection with E. cuniculi spores.
(A) Megasome (me) with amorphous and electron dense material and E. cuniculi spores (arrow) inside. Ultramicrography of macrophages from APerC BALB/c. (B) Megasome with amorphous and electrodense material, myelin figure (head arrow) and spores (arrow) in XID APerC by ultramicrography. (C) Homotypic phagosomes vacuoles in macrophages from APerC BALB/c by ultramicrography. (D) Degraded macrophage (M) containing amorphous material and involving spore of E. cuniculi in megasome (me) in XID APerC, by ultramicrography. (E) B-1 cells (B-1) and pre-B-1 CDP interaction with macrophage with a large megasome (me). Insert: Note the close relationship between macrophage membranes and B-1 cell.
Fig 3.
Phagocytic activity of BALB/c and XID mice adherent peritoneal cells infected with E. cuniculi.
The phagocytic capacity and phagocytic index were obtained from adherent peritoneal cells infected with E. cuniculi after 1, 48, and 96 h. Data are represented as mean ± SEM (*P < 0.05, ** P < 0.01, One way ANOVA with Tukey post-test).
Fig 4.
Necrosis and apoptosis of BALB/c and XID APerC cultures at 48 and 96 h.
Percentual of: (A) death cells. (B) apoptotic cells. (C) necrotic cells. The data are presented as mean ± SEM (*P < 0.05, ** P < 0.01, *** P <0.001, One way ANOVA with Tukey post-test).
Fig 5.
Relations between phagocytes from XID APerC and phagocytized E. cuniculi spores after 1 h of infection by ultramicrography.
(A) Contact area between the phagocytic vacuole membrane of the macrophages and the internalized spore wall (arrow). (B) Germinated spore with extruded polar tube (arrow) from the phagosome vacuole.
Fig 6.
Activation phenotype of macrophages from BALB/c or XID APerC infected with E. cuniculi spores.
(A) Total percentage of F4/80+CD11b+ macrophages. (B) Proportion of F4/80+CD11b+ macrophages expressing CD40+. (C) Proportion of F4/80+CD11b+ macrophages expressing CD206. (D) Mean fluorescence intensity (MFI) ratio of CD40/CD206 molecules expression on F4/80+CD11b+ macrophages. (E) Mean fluorescence intensity of CD80+ and CD86+ coestimulatory molecules on macrophages. The data are represented as mean ±SEM (* P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001, One way ANOVA with multiple comparisons and Bonferroni post-test).
Fig 7.
Cytokines levels in the supernatants of BALB/c and XID APerC at 30 minutes, 1 hour, 48 and 96 hours.
Data are represented as mean ±SEM (** P <0.01, *** P <0.001and ND = not detected, One-way ANOVA with multiple comparisons and Tukey post-test).
Fig 8.
Phagocytic activity of B-1 CDP culture after 1 h and 48 h of infection with E. cuniculi by ultramicrography.
(A) Pre-B-1 CDP (*) in communication between them and with E. cuniculi spores (arrow). Amplified view of the interaction between B-1 cells. (B) B-1 CDP with the presence of amorphous material inside phagocytic vacuoles (*). (C) Non-germinated spores (s) outside the cells with a thick wall composed of two layers and a plasma membrane. (D) Extracellular vesicles (head arrow) in the membrane of pre-B-1 CDP with the presence of amorphous material (*) inside. (E) Extracellular vesicles in membranes of pre-B-1 CDP (*) and B-1 cell.
Fig 9.
Necrosis and apoptosis, phagocytic activity and cytokines levels in the supernatants from B-1 CDP cultures.
(A) Percentual of death cells, apoptotic cells and necrotic cells in B-1 CDP culture at 48 and 96 h. (B) Phagocytic capacity and phagocytic index in B-1 CDP culture infected with E. cuniculi after 1, 48, and 96 h. (C) Cytokines levels after 1, 48, and 96 h post infection with E. cuniculi. The data are presented as mean ± SEM (*P < 0.05, ** P < 0.01, *** P <0.001, One way ANOVA with Tukey post-test, ND = not detected).