Figure 4.

Enhancement of IL-17A-producing γδT cells and left ventricle remodelling in TDAG8 KO mice after MI. (a) mRNA expression levels of the indicated genes in infarcted and non-infarcted areas of WT mice (sham n = 3, MI n = 3–7) and TDAG8 KO mice (sham n = 3, MI n = 3–5) 3 days after MI or sham operation. # indicates that IL-17a mRNA was not detected. (b) All cardiac γδT cells isolated from WT and TDAG8 KO mice were treated with IL-1β (10 ng/ml) and IL-23 (10 ng/ml) on an anti-CD3 antibody-coated plate. Four infarcted hearts were combined for this analysis. After 48 h, the amount of secreted IL-17A in the supernatants was measured using ELISA. ELISA assay was performed in triplicates. (c) Infiltrated IL-17A-producing T cells in the infarcted hearts of WT mice and TDAG8 KO mice 3 days after MI were further analysed for γδTCR and CD4 expression using flow cytometry. Five heart samples were combined as one sample and used for analysis. (d) TUNEL staining of the infarcted area 3 days after MI. Tissue sections from WT mice (sham n = 3, MI n = 3) and TDAG8 KO mice (sham n = 3, MI n = 3) 3 days after MI were stained with TUNEL (red) and DAPI (blue). Arrows indicate apoptotic cells. (e) mRNA expression levels of fibrosis-related genes in infarcted and non-infarcted areas of WT mice (n = 3) and TDAG8 KO mice (n = 3) 7 days after MI or sham operation. Total mRNA extracted from sham-operated ventricles or MI-operated ventricles (separated into infarcted and non-infarcted areas) were subjected to real-time RT-PCR. (f) Representative heart sections of infarct and non-infarct areas 7 days after MI in WT mice (n = 4) and TDAG8 KO mice (n = 3) stained with Picrosirius Red. (g) The fibrotic area was quantified. Comparisons were assessed with unpaired Student’s t-tests. Error bars represent the mean ± SEM (N.S., not significant).