Morphometric comparisons of rat alveolar macrophages, pulmonary interstitial macrophages, and blood monocytes

Abstract

Pulmonary interstitial macrophages (IM) account for a substantial fraction of the total pulmonary macrophage (PM) population in the mammalian lung, with the remaining balance of extravascular mononuclear phagocytes being mainly alveolar macrophages (AM). Unlike the AM that can be harvested readily by bronchoalveolar lavage, the lung's IM subpopulation of PM has been characterized less well, primarily because of its relative inaccessibility. Recently we developed a method to isolate viable IM from rat lungs using an Fc gamma receptor affinity technique in conjunction with multiparameter flow cytometry. Using this approach, we undertook the present investigation to characterize quantitatively the structural features of the IM and to compare the morphologic attributes of this subpopulation of PM to those of flow cytometrically sorted AM and blood monocytes (BM). Measured or calculated parameters for each population included mean cellular equivalent circular diameter, cell area and volume, and nuclear, mitochondrial, cytoplasmic, and lysosomal volume densities in each cell type. Lysosomal volume densities were subcategorized further into primary lysosomes, small secondary lysosomes, large secondary lysosomes, lipid droplets, and vacuoles. Additionally, the shape, form, and surface irregularity of the cells and various subcellular components were determined. Comparisons of the size and other structural features of the AM, IM, and BM have indicated that (1) the morphologic phenotypes of these three populations of mononuclear phagocytes distinctly differ from one another, (2) the IM and BM are morphologically and morphometrically more akin to one another than they are to AM, and (3) the IM are more similar to the AM than are the BM. These findings suggest that the IM may represent a transitional stage of maturation between BM and AM. Our findings, however, do not rule out the possibility that at least some of the lung's IM are a discrete, BM-independent population of macrophages.