Fig. 3: Ac-6-FP promotes fibrosis resolution via an impact on the MAIT macrophage dialog.
From: MAIT cell inhibition promotes liver fibrosis regression via macrophage phenotype reprogramming
a–d Mice were injected CCl4 or oil for 4 weeks along the protocol described in Fig. 2d and frequencies of resident Kupffer cells (KC) and MoMac analyzed after cessation of injury. a Frequency of intrahepatic KC, Ly6Clo and Ly6Chi MoMac in C57BL/6 J. Each point is the mean of five mice except for oil where n = 13 mice. b Frequencies of MR1+ cells among KC, Ly6Clo and Ly6Chi MoMac at day 1 after CCl4 cessation (n = 7 mice/group) vs oil (n = 6 mice/group). **p = 0.001. c Frequencies of Ly6Chi, Ly6Clo (pooled data from two experiments, n = 11 mice/group), and CCR2+ (n = 6 mice/group) MoMac from C57BL/6 J mice at day 1 after the last CCl4 injection. *p = 0.01, **p = 0.002. d Frequencies of Ly6Chi and Ly6Clo from MR1−/− mice exposed to Ac-6-FP or vehicle, at day 1 after CCl4 cessation (n = 5 mice/group) compared to oil (n = 6). **p = 0.009. e, f Representative images of immunofluorescent staining of CD206 and CCR2 and quantification of the CCR2/CD206 mean intensity ratio in BMDM alone or co-cultured with MAIT cells for 48 h in the presence of e Ac-6-FP or vehicle (n = 5 experiments except for BMDM + Ac-6-FP where n = 2; each point represent the mean per experiment; *p = 0.02 for BMDM + MAIT vs BMDM alone, and *p = 0.04 for BMDM + MAIT + Ac-6-FP vs BMDM + MAIT + vehicle), or f antibodies to TNFα and/or IL17 or control isotype (typical representative of two experiments; each point is one field; n = 12 fields/group except for BMDM + MAIT + isotype where n = 15 fields). *p = 0.02; **p = 0.002; ****p < 0.0001. Scale bar is 10 µm. g Timeline of CCl4, Ac-6-FP and clodronate or PBS liposome administration in C57BL/6 J mice created with BioRender® software. h Clodronate vs PBS liposome fold change of Ly6Chi and Ly6Clo MoMac frequencies at day 1, 2, and 4 after CCl4 cessation. Each point represents the mean value of 5 mice for day 1 and day 2, and 4 mice for day 4. i Representative images and quantification of Sirius red areas (typical representative of n = 4 experiments) of liver tissue sections from mice treated with Ac-6-FP or vehicle (n = 5 mice/group except for vehicle clodronate group where n = 4 mice). Sample were collected 4 days after the last CCl4 injection. Scale bar is 100 µm. *p = 0.05. Data are mean ± S.D. Statistical analysis was performed by (b–d, i) two-tailed Mann–Whitney test or (e, f) Kruskal–Wallis followed by Dunn’s multiple comparisons post-test. KC Kupffer cells, BMDM bone marrow-derived macrophages, clodro clodronate-encapsulated liposomes, PBS phosphate-buffered saline-encapsulated liposomes. Source data are provided as a Source Data file.
