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A. W. Taylor, C. Lau; Neural Retina Layer Regulation of Macrophage Functionality. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1992.
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We have reported that within the healthy retina we can identify the presence of suppressor macrophages, and that retinal pigment epithelial (RPE) cells can promote suppressor macrophage induction. Therefore, we asked can other tissue layers that makeup the posterior microenvironment of the eye, in this abstract the neural retina layer (NR), contribute to this regulation of macrophage functionality?
The NR from healthy C57BL/6J mouse eyes was dissected and placed in culture for 24 hours with serum-free media. The conditioned media (CM) was collected and used to treat resting or endotoxin-stimulated macrophages (J774A.1 cell line, and primary resident peritoneal exudate cells). After 48 hours, the treated macrophage culture supernatants were assayed by multiplex analysis for IL-1β, TNF-α, and IL-10. Nitric oxide generation by the macrophages was assayed by Griess reagent. In addition, we assayed NR-CM treated primary macrophages for anti-inflammatory and suppressor activity by adding antigen while treating the macrophages with NR-CM for 24 hours. The treated antigen-pulsed macrophages where washed, and antigen-specific primed T cells were added to the macrophage cultures. The T cells were assayed after 48 hours of incubation for apoptosis by TUNEL staining and flowcytometry, and the supernatant was assayed for IFN-γ by ELISA.
Similar to our previous findings with RPE-CM, the NR-CM suppressed IL-1β and TNF-α while promoting IL-10 production by the endotoxin-stimulated macrophages. However, unlike RPE-CM the NR-CM suppressed nitric oxide production by the stimulated macrophages. The NR-CM treated macrophage did suppress IFN-γ production by antigen-stimulated primed T cells, but induced apoptosis in T cells independent of antigen-stimulation.
The NR suppressed macrophage inflammatory activity; however, it differed from RPE mediated induction of suppressor macrophages in that the NR induced a general anti-inflammatory function in macrophages. Such results suggest that the mechanisms of immunosuppression within the retina involve soluble factors produced by both the neural retina and the RPE layers, and that it is the RPE layer contributing to the induction of suppressor macrophages.
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