CCR6 Deficiency Increases Infarct Size after Murine Acute Myocardial Infarction

Figures

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  Figure 1. CCR6 deficiency results in decreased cardiac function and increased infarction size after ischemia-reperfusion injury. (A) Study design: wild-type and Ccr6−/− mice were subjected to ischemia-reperfusion injury, echocardiography was performed 3 weeks after injury (n = 3). Image was created using Biorender.com (Accessed on 2 September 2021). (B) Survival curve after ischemia-reperfusion injury. (C) Ejection fraction in Ccr6−/− mice compared to wild-type controls. (D) Stroke volume in Ccr6−/− mice compared to wild-type controls. (E) End-systolic volume in Ccr6−/− mice compared to wild-type controls. (F) Infarction size in Ccr6−/− mice compared to wild-type controls. (G) Representative images of Gomori’s 1-step trichrome stain. * p < 0.05, ** p < 0.01.

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  Figure 2. CCR6 deficiency in the bone marrow results in increased infarction size after ischemia-reperfusion injury. (A) Study design of bone marrow transplantation experiments, where the mice were subjected to ischemia-reperfusion injury. Echocardiography was performed 3 weeks after injury. Image was created using Biorender.com (Accessed on 2 September 2021). (B) Ejection fraction in wild-type recipient mice receiving either wild-type or Ccr6−/− BM (n = 4–6) and Ccr6−/− recipient mice receiving either wild-type or Ccr6−/− BM (n = 3). (C) Infarction size in wild-type recipient mice receiving either wild-type or Ccr6−/− BM (n = 4–6) and Ccr6−/− recipient mice receiving either wild-type or Ccr6−/− BM (n = 3). (D) Representative images of heart sections stained with Gomori’1 step trichrome staining of wild-type recipient mice receiving either wild-type or Ccr6−/− BM and Ccr6−/− recipient mice receiving either wild-type or Ccr6−/− BM. * p < 0.05.

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  Figure 3. Deficiency of CCR6 results in the increased infiltration of inflammatory immune cells in the cardiac tissue after ischemia-reperfusion injury. (A) Quantification of the number of myofibroblast in the scar from Ccr6−/− mice compared to wild-type mice 21 days after ischemia-reperfusion, including representative images of smooth-muscle actin (Green) staining. Scale bar = 50 µm. (B) Quantification of the number of vessels in the scar from Ccr6−/− mice compared to wild-type mice 21 days after ischemia-reperfusion, including representative images of CD31 (Red) staining. Scale bar = 50 µm. (C) Quantification of infiltrating B cells, 2 days after ischemia-reperfusion in Ccr6−/− mice compared to wild-type mice, including representative images of B220 (Yellow) staining. Scale bar = 50 µm. (D–F) Quantification of infiltrating inflammatory MPO+ immune cells (D), inflammatory macrophages (E) and reparatory macrophages (F), 2 days after ischemia-reperfusion in Ccr6−/− mice compared to wild-type mice. (G) Representative images of MPO (Red) and Mac3 (Green) staining. All n = 3. * p < 0.05, ** p < 0.01.

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  Figure 4. Deficiency of CCR6 results in increased apoptosis, especially in cardiomyocytes, after ischemia-reperfusion injury. (A–C) Representative images of TUNEL (Green) staining (A) and TUNEL (Green)/Troponin (Red) double staining (B), including the quantification of total TUNEL+ cells (C), 2 days after ischemia-reperfusion in Ccr6−/− mice compared to wild-type mice. Scale bar = 50 µm. (D–F) Representative images of Ki67 (Green) staining (D) and Ki67 (Green)/smooth-muscle actin (Red) double staining (E), including the quantification of total Ki67+ cells (F), 2 days after ischemia-reperfusion in Ccr6−/− mice compared to wild-type mice. Scale bar = 50 µm. Arrows point at positively stained cells. All n = 3.