Determination on IDD activity of human meninx fibroblast with HPLC

  Objective   To determine indoleamine 2, 3-dioxygenase(IDO)activity in human meninx fibroblast(HMF)by using reversed-phase HPLC and establishing a simple and reliable method, and to explore clinical significance of IDO.

  Methods   IDO activity is indirectly determined by detecting the amount of tryptophan and kynurenine with FLD and DAD respectively.The ability to restrict growth of T.gondii was evaluted by ³H-U incroporation assay.

  Results   The average coefficients of recovery of tryptophan an kynurenine were 99.02% and 102.18% respectively, their coefficients of variation were 1.03% and 1.89% respectively, and their minimal detected concentration were 8.77 ng/ml and 32.75 ng/ml respectively.IDO induced by IFN-γ could degrade the intracellular tryptophan to restrict T.gondii growth.

  Conclusion   Determination of IDO activity by using reversed-phase HPLC was a sensitive, simple and reliable method.IDO could restrict T.gondii growth.