Isolation of rabbit aqueous humor-derived exosomes and their immunosuppression function.
- Author:
Zhi-xiong LIAO
1
;
Zheng-qing LOU
;
Zhong XIAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Aqueous Humor; immunology; Cells, Cultured; Exosomes; immunology; metabolism; ultrastructure; Female; Immune Tolerance; Male; Rabbits; T-Lymphocytes; immunology
- From: Journal of Zhejiang University. Medical sciences 2012;41(3):315-319
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo isolate exosomes from rabbit aqueous humor and to investigate their immunosuppression function.
METHODSAqueous 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.
RESULTSEight 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.
CONCLUSIONImmunosuppressive exosomes can be isolated from rabbit aqueous humor, which may be involved in immunotolerance of the eye.
