Purpose: To compare the basic proteomic composition of aqueous humour (AH) fro m patients with corneal rejection (patients) with AH from patients with cataract (controls). Methods: Aqueous humour was analysed for total protein concentratio n using Bradford’s method and for protein composition using two-dimensional (2 D) gel electrophoresis. Image analysis was used to detect protein spots in 2D ge ls that were increased by more than factor 2 in patients as compared with contro ls. Increased spots were identified by immunoblotting and mass spectrometry. Res ults: Aqueous humour from patients contained significantly higher total protein concentration than did AH from controls. A total of 31 spots were significantly increased in 2D gels from patients. The spots were derived from albumin, α1-an titrypsin, apolipoprotein J, cytokeratin type II, serin proteinase inhibitor and transthyretin. After correction of spot volumes by total protein concentrations , 10 spots derived from albumin, cytokeratin type II and α1-antitrypsin remain ed significantly increased. Conclusion: The proteomic composition of AH differed significantly between patients and controls. The identified proteins suggest th at the changes in AH are due to at least three different mechanisms: breakdown o f the aqueousblood barrier, enzymatic degradation, and liberation of locally syn thesized proteins.
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