Figure 1
Figure 1. Tlr3−/− mice are highly susceptible to pulmonary aspergillosis. C57BL/6 or Tlr3−/− mice were infected intranasally with live A fumigatus conidia (6-8 mice/group). (A) Fungal growth (mean CFUs ± SE) in the lungs and brains of infected mice was assessed at different days postinfection (dpi). (B) Lung histology (periodic acid-Schiff staining), bronchoalveolar lavage morphometry, expressed as a percentage (mean ± SD) of mononuclear cells (MNCs) or polymorphonuclear (PMN) cells, and immunofluorescence of infected mice at different dpi. Note the sustained inflammatory cell recruitment in the lungs and bronchoalveolar lavage (May-Grünwald Giemsa staining in the inset using a 100×/1.3 oil objective) of Tlr3−/− mice and the presence of peribronchiolar lymphocyte infiltrates (at 3 dpi; arrows in periodic acid-Schiff staining and immunofluorescence images). Cell-surface markers used for DC and T-cell visualization were CD11c (green, Alexa Fluor 488) and CD3 epsilon followed by PE-secondary Ab (Red-647), respectively. Cell nuclei were stained blue with 4′,6-diamidino-2-phenylindole and then mounted with Vectashield mounting medium (Vector Laboratories). Representative images of 2 independent experiments were visualized with a 40×/0.75 objective. All images were captured on the BX51 upright microscope (Olympus) using a high-resolution Microscopy Olympus DP71 camera (Olympus) and digitally acquired using the Cell∧P software (Version 3.2 build 2108, Olympus). Bars in histology sections indicate magnifications. (C) Lung total RNA from infected mice was extracted from naive (0) or at 1, 3, and 7 dpi, and the relative expression of Cxcl1, Cxcl2, Mpo, Il1b, Ifna1, and Ifnb1 was assessed by RT-PCR. (D) Total RNA was extracted from purified CD4+ T cells from the draining lymph nodes of naive or infected mice at 7 dpi, and the relative expression of Tbet, Rorc, and Foxp3 was assessed by RT-PCR. (E) Total lung homogenates at 3 and 7 dpi were assessed for levels of IFN-γ, IL-17A and IL-10 by specific ELISA (mean values ± SD). Data are pooled from 3 independent experiments. *P < .05; **P < .01; and ***P < .001 for infected versus naive mice (panel D) or Tlr3−/− vs C57BL/6 mice panel (panels A, C, and E).

Tlr3−/− mice are highly susceptible to pulmonary aspergillosis. C57BL/6 or Tlr3−/− mice were infected intranasally with live A fumigatus conidia (6-8 mice/group). (A) Fungal growth (mean CFUs ± SE) in the lungs and brains of infected mice was assessed at different days postinfection (dpi). (B) Lung histology (periodic acid-Schiff staining), bronchoalveolar lavage morphometry, expressed as a percentage (mean ± SD) of mononuclear cells (MNCs) or polymorphonuclear (PMN) cells, and immunofluorescence of infected mice at different dpi. Note the sustained inflammatory cell recruitment in the lungs and bronchoalveolar lavage (May-Grünwald Giemsa staining in the inset using a 100×/1.3 oil objective) of Tlr3−/− mice and the presence of peribronchiolar lymphocyte infiltrates (at 3 dpi; arrows in periodic acid-Schiff staining and immunofluorescence images). Cell-surface markers used for DC and T-cell visualization were CD11c (green, Alexa Fluor 488) and CD3 epsilon followed by PE-secondary Ab (Red-647), respectively. Cell nuclei were stained blue with 4′,6-diamidino-2-phenylindole and then mounted with Vectashield mounting medium (Vector Laboratories). Representative images of 2 independent experiments were visualized with a 40×/0.75 objective. All images were captured on the BX51 upright microscope (Olympus) using a high-resolution Microscopy Olympus DP71 camera (Olympus) and digitally acquired using the Cell∧P software (Version 3.2 build 2108, Olympus). Bars in histology sections indicate magnifications. (C) Lung total RNA from infected mice was extracted from naive (0) or at 1, 3, and 7 dpi, and the relative expression of Cxcl1, Cxcl2, Mpo, Il1b, Ifna1, and Ifnb1 was assessed by RT-PCR. (D) Total RNA was extracted from purified CD4+ T cells from the draining lymph nodes of naive or infected mice at 7 dpi, and the relative expression of Tbet, Rorc, and Foxp3 was assessed by RT-PCR. (E) Total lung homogenates at 3 and 7 dpi were assessed for levels of IFN-γ, IL-17A and IL-10 by specific ELISA (mean values ± SD). Data are pooled from 3 independent experiments. *P < .05; **P < .01; and ***P < .001 for infected versus naive mice (panel D) or Tlr3−/− vs C57BL/6 mice panel (panels A, C, and E).

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