Immune response to intra-ocular injection of particulate ß-(1,3)(1,6) glucan

Description of cellular and molecular landscape of vitreal immune cells following optic nerve crush injury plus intra-ocular injection of particulate ß-glucan after 3days, in WT and Itgam-/- mice Overall design: Intro-ocular (i.o.) of highly purified particulate ß-(1,3)(1,6) glucan, prepared from s. cerevisiae, triggers a strong immune response in the vitreal humor and the retina. When combined with retro-orbital optic nerve crush injury, ß-(1,3)(1,6) glucan elicited ocular inflammation enables robust regenerative axon growth of injured retinal ganglion cells. We observed that in mice deficient for the MAC-1 receptor component integrin alphaM (encoded by Itgam), i.o. ß-(1,3)(1,6) glucan results in enhanced RGC axon regeneration when compared to parallel processed WT mice (also treated with i.o. ß-(1,3)(1,6) glucan). For a comparative analysis of ocular immune infiltrates at 3days post optic nerve crush plus ß-(1,3)(1,6) glucan, we used magnetic beads sorting (anti-CD45 MACS) of all leukocytes in the vitreous and retina. Leukocytes from WT and Itgam-/- eyes were prepared for scRNA-sequencing and processed separately.