Figure 5. Absence of TRIF results in reduced production of myeloid cell-recruiting factors, except in mice exhibiting breakthrough inflammation.

Two weeks following rAAV injection, C57BL/6J and TRIF^−/− mice received PBS or DT and were sacrificed 48 hours later. (A) Fold change gene expression in total liver, hepatocytes, LSECs, and KCs, + PBS group and + DT group compared to the + PBS group median, Hprt was used as the reference gene. (C) mRNA expression relative to Hprt for Ccl7 in hepatocytes and LSECs as non-detects in the + PBS groups complicated fold change calculations. (D) Serum protein levels for CCL2 and CXCL1. Correlation of serum ALT with (B) gene expression fold change levels in total liver, (E) serum chemokines, and (F) monocyte and neutrophil infiltrate for B6 + DT (black) and TRIF^−/− + DT (white) groups. (A–G) Data are combined for 2–3 experiments. Each data point represents an individual mouse, bars represent the median. For (A–C, F) total liver and hepatocytes n = 8–15 per group and for LSECs and KCs n = 5–10 per group. For (D, E) serum: n = 10, B6 + PBS; n = 9, B6 + DT; n = 8, TRIF^−/− + PBS; n = 13, TRIF^−/− + DT. (A) Ccl2 has one non-detect value in each + PBS group not noted on the graph. Significance determined by (A) an individual Mann-Whitney test per population, (B, E, F) Goodness of Fit, with R² and p-value reported on the graph, and (D) by four pairwise Mann-Whitney tests and the p-values adjusted for multiple comparisons using the Holm-Bonferroni method as described in the methods. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; non-significance represented by absence of bar. B6, C57BL/6J; DT, diphtheria toxin; Hepa; hepatocyte; KC, Kupffer cell; LSEC, liver sinusoidal endothelial cell; TRIF, TIR-domain-containing adapter-inducing interferon-β; MyD88, myeloid differentiation primary-response protein 88; and ND, non-detect.