Article Figures & Data
Figures
- FIGURE 1.
a, Effect of CL2MBP liposomes on AMs 2 days after i.n. administration. The bars indicate the percentage of total AMs remaining in AM− mice (▪) or AM+ (liposomes) mice (▨) compared to mice receiving saline (□; 100%). The data are presented as mean and SEM from five mice per group. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice. b, Apoptotic AMs were visualized by immunostaining lung tissue for cleaved PARP 7 h after CL2MBP liposome treatment (original magnification, ×80).
- FIGURE 2.
Effect of AM depletion on survival of mice following M. tuberculosis infection. BALB/c mice (n = 10 per group) were i.n. administered with saline, liposomes, or CL2MBP liposomes prior to and after bacterial challenge of 1 × 105 M. tuberculosis H37Rv. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice; ‡, p < AM+ (saline) mice vs AM+ (liposomes) mice.
- FIGURE 3.
Growth of M. tuberculosis in lungs (a) and liver (b) of AM+ (saline/liposomes) mice and AM− mice. Data are mean and SEM of CFUs from eight mice per group for each time point. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice.
- FIGURE 4.
Lung histopathology. a, Two weeks postinfection, lungs of AM+ mice showed well-shaped granulomas, chiefly composed by macrophages and few lymphocytes (hematoxylin and eosin staining; original magnification, ×50). b, At the same time point, lungs of AM− mice displayed slight perivascular lymphocytic infiltrates and degenerated macrophages in alveolar spaces. Well-defined granulomas were not found (hematoxylin and eosin staining; original magnification, ×50). c, Five weeks postinfection, lungs of AM+ mice displayed a diffuse inflammatory infiltrate predominantly composed of macrophages (hematoxylin and eosin staining; original magnification, ×25). d, Lungs of AM− mice presented an almost normal aspect (hematoxylin and eosin staining; original magnification, ×25). The slides shown are representative for a total of eight mice per group for each time point.
- FIGURE 5.
The effect of AM depletion on M. tuberculosis-mediated induction of type 1 cytokines (a) and type 2 cytokines (b) in lungs of AM+ (saline) mice (□), AM+ (liposomes) mice (▨), and AM− mice (▪) 2 wk postinfection. The data represent the mean and SEM of eight mice. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice.
- FIGURE 6.
Splenocytes from infected AM− mice (▪) release more IFN-γ in response to PPD and anti-CD3/28 Abs and less IL-4 in response to anti-CD3/28 ABS than splenocytes from infected AM+ mice (□, saline; ▨, liposomes). Splenocytes were harvested 2 wk after i.n. inoculation with M. tuberculosis, and stimulated for 48 h. The data are mean and SEM of eight mice per group. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice; ‡, p < AM+ (saline) mice vs AM+ (liposomes) mice.
Tables
Cells (×104/ml) CD4+ CD8+ CD4+/CD69+ CD4+/CD25+ CD8+/CD69+ Gr-1+ 2 wk postinfection AM+ (saline) 356 ± 63.9 69.9 ± 1 31.1 ± 0.6 2.8 ± 1 6.6 ± 0.9 1.2 ± 0.5 57 ± 5.4 AM+ (liposomes) 265.7 ± 50.4 65.6 ± 1.2‡ 33.3 ± 1.1 3.4 ± 1.1 6.3 ± 0.6 1 ± 0.5 63 ± 3.9 AM− 444.5 ± 111.6 63 ± 5.3* 34.1 ± 2*† 12.4 ± 1.5*† 13.1 ± 1.7*† 5.4 ± 0.2*† 58.3 ± 3.1 5 wk postinfection AM+ (saline) 225 ± 26 53.3 ± 2.4 42.9 ± 1.7 4.2 ± 0.6 2.6 ± 0.6 2.6 ± 1 56.6 ± 5.2 AM+ (liposomes) 207.8 ± 11.5 52 ± 1.3 39.1 ± 2.9 8.4 ± 1‡ 2.3 ± 0.4 9.6 ± 1.2‡ 64.5 ± 4.1 AM− 136.5 ± 15.1*† 53.9 ± 1 38.1 ± 2.6 6.8 ± 0.3* 1.6 ± 0.4 7.8 ± 0.7* 54.8 ± 7.4 -
a Cell subsets in the lungs of mice infected with M. tuberculosis, 2 and 5 wk postinfection. FACS analysis was performed on pooled cells from two mice for each analysis from a total of eight mice per group for each time point as described in Materials and Methods. FACS results are expressed as the percent of CD4+, CD8+, CD25+, and CD69+ within the CD3+ population or the percent of Gr-1+ within the PMN population.
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b , p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice; and ‡, p < 0.05 AM+ (saline) vs AM+ mice (liposomes).
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Cells (×104/ml) AM (×104/ml) PMN (×104/ml) Lymphocytes (×104/ml) 2 wk postinfection AM+ (saline) 79 ± 7.2 32.9 ± 5.7 28.8 ± 6.1 17.2 ± 1 AM+ (liposomes) 63.8 ± 7.5 31.8 ± 1.7 19.5 ± 6.4 14 ± 3.4 AM− 111 ± 7.3*† 16.3 ± 2.9*† 53.6 ± 4.2*† 41 ± 5*† 5 wk postinfection AM+ (saline) 308 ± 17.0 44 ± 3.4 182 ± 14 82.8 ± 16.7 AM+ (liposomes) 196 ± 27.6‡ 27.2 ± 6.1‡ 120 ± 23.4 54.6 ± 10.5 AM− 166 ± 16.7* 14.6 ± 1.5* 132.5 ± 13.9* 43 ± 5.5* -
a Leukocytes in BALFs of mice infected with M. tuberculosis, 2 and 5 wk postinfection. Cells from four mice were counted and stained with hematoxylin and eosin. ∗, p < 0.05 AM− mice vs AM+ (saline) mice; †, p < 0.05 AM− mice vs AM+ (liposomes) mice; and ‡, p < 0.05 AM+ (saline) vs AM+ mice (liposomes).
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