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Inhibition of T Cell Proliferation by Combined Low-Tryptophan Medium and Tryptophan Metabolites in Vitro

Author: ZhangShuJian
Tutor: LuoYuDong；DaiXiangChen
School: Tianjin Medical University
Course: Surgery
Keywords: Low tryptophan medium Tryptophan metabolites Dendritic cells Inhibition of proliferation Apoptosis Immune tolerance
CLC: R392
Type: Master's thesis
Year: 2010
Downloads: 38
Quote: 0
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Abstract


Objective: This topic application of mouse bone marrow-derived dendritic cells (Dendritic Cell, DC) to stimulate allogeneic mouse T lymphocytes, the application of low tryptophan medium and tryptophan metabolites inhibit T lymphocyte proliferation , and reveals inhibit T lymphocyte proliferation mechanism . Methods: medium cytokine selection in vitro culture of mouse bone marrow-derived DC , DC surface antigens of CD80 by flow cytometry , CD86, of MHC II , of CD11c expression in low tryptophan medium added tryptophan metabolites mixed lymphocyte culture and to observe the impact of allogeneic T lymphocyte proliferation . Determination of T cell apoptosis using Annexin-V and PI double staining . Using the SPSS 15.0 software package for data analysis . Results: After in vitro culture , available to each mouse 5-6 × 106 bone marrow-derived DC , cells with typical dendritic structure , high expression of CD80 ( 94.56 % ) , CD86 ( 88.42 % ) of MHC- II ( 86.12 % ), CD11c (96.79%). Separable by immunomagnetic beads purity of more than 90% of CD4 T cells , mixed lymphocyte culture , low tryptophan group than normal tryptophan group stimulation index (SI) was significantly lower (P lt ; 0.001) , by adding The the tryptophan metabolic product of KYN group ,3-HK group ,3-HAA group SI values ??with concentration increased significantly reduced , and a significant difference ( P lt; 0.001 ) compared with the control group . Wherein 3-HAA inhibiting proliferation of CD4 T cells is the most obvious, and the lowest concentrations required . Low tryptophan medium with tryptophan metabolites CD4 T cell proliferation inhibition has superimposed . Annexin-V and PI double staining method for the determination of CD4 T cells apoptosis low tryptophan group apoptosis rate was significantly higher , and join the the tryptophan metabolites apoptosis rate further increased ( P lt ; 0.001 ) . Conclusion: The medium cytokines selection method can harvest a large number of bone marrow-derived DC , immunomagnetic beads separable purity higher CD4 T cells . Low tryptophan medium and tryptophan metabolites could inhibit the proliferation of CD4 T cells , and both have additive effect . Its cause CD4 T cell proliferation inhibition mechanism for \This provided the experimental basis for further study of immune tolerance after organ transplantation .

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