The aim of this study was to investigate the developmental expression of major histocompatibility complex class II (MHCII) by microglia and macrophages and their relationship to blood vessels in the retina, a representative tissue of the central nervous system. Such information is crucial to understanding the role of these cells in immune surveillance. Wholemount preparations of retinas from late embryonic, postnatal and adult rabbits were subjected to three-colour fluorescence microscopy using β2 integrin (CD18) and MHCII antibodies and biotinylated Griffonia simplicifolia B4 isolectin labelling of blood vessels. CD18+ cells consistently exhibited characteristics of macrophages or microglia in the vascularized and non-vascularized regions of the retina, respectively. At all ages, MHCII was expressed by a high proportion of cells in the vascularized region, which contained macrophage-like ‘parenchymal cells’ as well as typical perivascular macrophages. MHCII expression by ramified microglia, first detected on postnatal day 30, was lower in the peripheral retina and intermediate in the avascular region of the myelinated streak. The observed localization of MHCII+ cells in relation to blood vessels and location-dependent differences in MHCII expression point to the possibility that these cells may be distributed strategically within the retina to provide multiple lines of defence against immune challenge arriving via the retinal vasculature.