Supplementary Material for: Mesenchymal stromal cells from human Wharton’s jelly modulate the intraocular immune response in a glucocorticoid hypertension model: an exploratory analysis

Introduction: Glaucoma is a neurodegenerative disease characterized by the loss of retinal ganglion cells. Recent research suggests that immunological changes such as cytokine imbalance may play a role in its pathophysiology. This implies that immunomodulation, similar to that produced by mesenchymal cells, could be a potential therapeutic avenue for this disease. However, the effects of intravitreal injections of human Wharton’s jelly derived mesenchymal stromal cells(hWJ-MSCs) on intraocular immune response have not been assessed in ocular hypertension (OH) models. Methods: We measured explored this by measuring cytokine levels and expression of other markers, such as glial fibrillary acidic protein (GFAP) and T cells, in 15 randomly divided New Zealand rabbits: G1: OH; G2: hWJ-MSCs; and G3: OH+hWJ-MSCs. We analyzed the aqueous humor (IL-6, IL-8, and TNF-α) and vitreous humor (IFN-γ, IL-10, and TGF-β) using ELISA and flow cytometry (cell populations), as well as TCD3+, TCD3+/TCD4+, and TCD3+/TCD8+ lymphocytes, and GFAP in the retina and optic nerve through immunohistochemistry. Results: We found a decrease in TNF-α, IL-6, IFN-γ, IL-10, and IL-8 in G3 compared to G1 and an increase in TGF-β in both G2 and G3. TCD3+ retinal infiltration in all groups was primarily TCD8+ rather than TCD4+ cells, and strong GFAP expression was observed in both the retina and optic nerves in all groups. Conclusion: Our results suggest that cellular and humoral immune responses may play a role in glaucomatous optic neuropathy, and that intravitreal hWJ-MSCs can induce an immunosuppressive environment by inhibiting proinflammatory cytokines and enhancing regulatory cytokines.