Defects in intracellular trafficking of fungal cell wall synthases lead to aberrant host immune recognition
Fig 10
Macrophage activation in response to mar1Δ requires members of the CLR and TLR families.
BMMs were harvested from the indicated mouse strains (C57BL/6 background, unless otherwise noted) and co-incubated with C. neoformans (Cn) cells at an MOI of 10:1, Cn:BMMs, followed by quantification of TNF-α (pg/ml) in the supernatant by ELISA. (A) Card9 is involved in macrophage activation by mar1Δ cells. Data represent means of 3 replicates from 3 independent experiments (n = 9). **, p < 0.01 WT vs. Card9-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (B) MyD88 is required for TNF-α production by macrophages in response to mar1Δ. Data represent means of 3 replicates from 3 independent experiments (n = 9). ****, p < 0.0001 WT vs. MyD88-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (C) Dectin-1 is involved in the response to mar1Δ cells. Data represent means of 3 replicates from 2 experiments (n = 6). ****, p < 0.0001 WT vs. Dectin-1-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (D) TLR2/4-/- BMMs do not respond to mar1Δ cells. Data represent means of 3 replicates (n = 3). ****, p < 0.0001 WT vs. TLR2/4-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (E) TLR4 is not required for the production of TNF-α induced by mar1Δ cells. BMMs were isolated from C3H/HeJ mice with a null mutation in TLR4, and C3H/HeOuJ control mice. Data represent means of 3 replicates from 2 independent experiments (n = 6). **, p < 0.01 C3H/HeOuJ vs. C3H/HeJ BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test. (F) Macrophage activation by mar1Δ is partially dependent on TLR2. Data represent means of 3 replicates from 3 independent experiments (n = 9). ****, p < 0.0001 WT vs. TLR2-/- BMMs as determined by two-way ANOVA with Sidak’s multiple comparisons test.