Immunohistochemical characterization and morphometric analysis of macrophages in rat mammary tumors

Macrophages are an important leukocyte component of the microenvironment of neoplasms. Macrophages have classically been subdivided into M1 and M2, depending on their roles in immune response, wound healing, and promotion or inhibition of tumor growth. In human breast cancer, increased presence of M2 macrophages has been associated with poor prognosis. The authors hypothesized that rat mammary carcinomas have increased macrophage influx compared to benign mammary proliferative lesions and normal mammary glands as well. In humans, both M1 and M2 macrophages express CD68, while CD163 is expressed primarily by M2 macrophages. Based on a single immunolabeling protocol with anti-CD68 and anti-CD163 antibodies, the extent of macrophage influx was investigated by morphometry to quantitate the immunopositive cells in normal rat mammary glands, benign mammary proliferative lesions, and mammary carcinomas. In mammary carcinomas, there was significantly higher percentage of CD68+ cells compared to benign mammary proliferative lesions and normal mammary glands. There was also higher percentage of CD163+ cells in mammary carcinomas compared to benign mammary proliferative lesions. Thus, increase in CD68+ and CD163+ macrophages corresponded to increased malignancy of rat mammary tumors in this study.