[Isolation of rabbit aqueous humor-derived exosomes and their immunosuppression function].
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.


CONCLUSIONS
Immunosuppressive exosomes can be isolated from rabbit aqueous humor, which may be involved in immunotolerance of the eye.
Hangzhou Traditional Chinese Medicine Hospital, Hangzhou 310007, China. liaozx611@163.com
Zhi-xiong
ZX
Liao
Zheng-qing
ZQ
Lou
Zhong
Z
Xiao
D000818:Animals; D001082:Aqueous Humor / Q000276:immunology*; D002478:Cells, Cultured; D055354:Exosomes / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D007108:Immune Tolerance; D008297:Male; D011817:Rabbits; D013601:T-Lymphocytes / Q000276:immunology