Modulation of Metabolism Inflammation

To assess whether the previous observed findings were clearly due to CCR3-expressing hematopoietic cells, we generated mixed BM chimeras by transferring whole BM of apoE-/-CD45.1 mice to irradiated apoE-/-CCR3-/-CD45.2 recipient animals and BM of apoE-/-CCR3-/-CD45.2 mice to myeloablated apoE-/-CD45.1 recipient animals. The confirmation of the BM transplant success was performed by flow cytometry based on the circulating levels of CD45.1+ leukocytes in apoE-/-CCR3-/-CD45.2 mice and CD45.2+ circulating cells in apoE-/-CD45.1 animals (Columns D, E, F). Then, both mice chimeras were subjected to a high-fat or control diet for two months. When both chimeric animals were placed for two months to a high fat diet, the atheroma plaque analysis revealed that again, both chimeric BM transplanted animals, showed increased lesions in the aortic arch, the thoracic aorta and the aortic sinus when they were fed with a high-fat diet for two months (Columns J, K and L, respectively).  The atheroma in apoE-/-CD45.1 mice transplanted with BM from apoE-/-CCR3-/- animals was significantly greater than that developed in apoE-/-CCR3-/- mice transplanted with BM from apoE-/-CD45.1 animals (Column L). Further examination of the cellularity of the atherosclerotic lesion, the number of infiltrated macrophages (Mac-3+ cells) and CD8+ T lymphocytes, were significantly higher in apoE-/-CD45.1 mice transplanted with BM from apoE-/-CCR3-/- animals fed with the same diet than in apoE-/-CCR3-/- mice transplanted with BM from apoE-/-CD45.1 animals (Columns M and N). Regarding the blood analysis by flow cytometry, a decrease in the percentage of CD4+ and CD8+ lymphocytes was observed when the diet was administered, independent on the genotype (Columns E and F). However, a significant increase in the percentage of CD115+ monocytes was detected in BM apoE-/- CCR3-/- mice compared to BM apoE-/- CD45.1 mice fed an atherogenic diet (Column D). Bone marrow cytometry analysis of these leukocyte subpopulations did not reveal any significant changes between the groups studied (Columns G, H, I)