Characterization of testicular macrophage subpopulations in mice

Abstract

Testis is an immune privileged site, a feature that prevents germ cells from eliciting an autoimmune response. Macrophages contribute to this state of tolerance by adopting an immunoregulatory phenotype. Here, we further characterized their features in mice by analyzing surface markers, anatomic localization as well as morphology and function. Testicular macrophages (TMΦ) were stained for various surface receptors, and MHCII and CD206 were found to be most suitable to discriminate between two subpopulations. Our immunohistochemical analysis further confirmed a predominant localization of CD206⁺ cells in the interstitial space. Imaging flow cytometry revealed that both subtypes of TMΦ differed in size and contrast, and to some extent also in their ability to engulf high-molecular dextran. To investigate whether the polarization of the immune system had any influence on the phenotype of TMΦ, we compared C57BL/6 and BALB/c mice. Importantly, our analysis revealed that the abundance of cells expressing either MHCII or any of the scavenger receptors CD206, CD163 and CD71 differed between both mouse strains. In addition, the presence of the glucocorticoid receptor in macrophages affected the ratio between individual subpopulations, which is consistent with a crucial role of glucocorticoids in macrophage polarization. Collectively, our results indicate that TMΦ are composed in a variable ratio of distinct subsets with characteristic features, which may shape the immune privilege of the testis also in humans.

Keywords: Glucocorticoids; Imaging flow cytometry; Macrophages; Mice; Testis.