Ultrastructural analysis of a specific hamster alveolar macrophage antigen

Alveolar macrophages of hamsters have a cell surface antigen not found on other pulmonary cells or macrophages from other sites. A specific mouse monoclonal antibody and protein A-colloidal gold were used to study the distribution of this antigen at the ultrastructural level. By quantitating the number of antigenic sites per unit linear distance of membrane of the total cell surface, it was established that this antigen is randomly distributed on the plasma membrane. Cell-to-cell heterogeneity in antigen density was correlated with other ultrastructural features of lung macrophages studied morphometrically. A strong correlation (r = 0.77) between antigen density and secondary lysosome content was found, supporting the thesis that expression of this antigen increases with macrophage age. No apparent topographic relationship of surface antigen to any cytoplasmic organelles could be inferred from the ultrastructural morphology. This study also used immuno-ultrastructural techniques to visualize stages of the internalization of labeled antigen in live cells. While some cells retained a random distribution of antigen at 0 degrees C, most showed movement of antigenic sites away from pseudopodia to clusters on more linear (less ruffled) portions of cell surface. At 37 degrees C, the antigen was internalized and could be visualized within coated pits and endocytic vacuoles. Definition of the anatomic distribution of this antigen under various conditions provides a baseline for further ultrastructural investigations of the function of this antigen.