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  • Title: [Isolation of rabbit aqueous humor-derived exosomes and their immunosuppression function].
    Author: Liao ZX, Lou ZQ, Xiao Z.
    Journal: Zhejiang Da Xue Xue Bao Yi Xue Ban; 2012 May; 41(3):315-9. PubMed ID: 22723168.
    Abstract:
    OBJECTIVE: To isolate exosomes from rabbit aqueous humor and to investigate their immunosuppression function. METHODS: Aqueous humor was collected from 40 New Zealand rabbits and exosomes were isolated by fractional separation and ultracentrifugation methods; the morphology was studied with electron microscopy. The immunosuppressive-related proteins of exosomes were detected with Western blotting; their inhibitory effect on ConA-induced proliferation of T lymphocyte was estimation with CCK-8 cells proliferation assay. RESULTS: Eight milliliters of aqueous humor were collected from 40 New Zealand rabbits and 200 μg exosomes was yielded. Under electron microscope, the exosomes had typical structure of lipid bi-layer with a diameter of 50-100 nm. The results of Western blotting showed that these exosomes expressed Hsp70, CD9 and Alix but not Grp94, presenting a typical exosomes protein profile. Moreover, exosomes expressed high level of TGF-β and significantly inhibited the proliferation of T lymphocytes. CONCLUSION: Immunosuppressive exosomes can be isolated from rabbit aqueous humor, which may be involved in immunotolerance of the eye.
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