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Suppression of CD8~+T Cells by Regulatory Dendritic Cells

Author: WangZuo
Tutor: SongWenGang；ZhangMingHui
School: Taishan Medical College
Course: Immunology
Keywords: Stromal cells in the liver CD8 ~ T cells Regulatory dendritic cells Immunomodulatory Nitric oxide
CLC: R392
Type: Master's thesis
Year: 2010
Downloads: 39
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Abstract

The purpose of the use of liver stromal cells (liver stromal cells, LSCs) mature dendritic cells (mature dendritic cells, mDCs) induced to differentiate into regulatory dendritic cells (regulatory of dendritic cells DCreg) explore DCreg antigen-specific CD8 T cells the immune negative regulation. 1 immunomagnetic bead separation technology separation of C57BL / 6 mice bone marrow-derived mDCs, LSCs induced after DCreg MHC class I molecules restricted OVA257-264 antigen peptide-specific TCR transgenic OT1 mouse CD8 T cells. 2. Flow cytometry DCreg, mDCs immature dendritic cells (immature dendritic cells, imDC) expression of cell surface molecules, antigen-specific CD8 T cell activation, proliferation and cytokine secretion. 3 of CFSE marker to detect antigen-specific CD8 T cell division. RT-PCR to detect liver stromal cell expression of cytokines and chemokines. 5 Following reinfusion DCreg, mDCs and CD8 T method detection in vivo DCreg of inhibition of CD8 T cells. 6. Griess reagent cartridge detecting the NO content in the culture supernatant. Results established stable LSCs with mDCs co-culture system and generated induced DCreg. LSCs induction of DCreg of in vivo and in vitro can inhibit the proliferation of antigen-specific CD8 T cells. In vivo and in vitro experiments according to the following processing is divided into four groups: CD8 T control group, mDCs CD8 T group, DCreg CD8 T group, mDCs CD8 T-DCreg group. The results showed that in vitro after 72 hours of incubation able to inhibit antigen-specific induced by mDCs DCreg CD8 T cell proliferation., But DCreg does not affect the activation of antigen-specific CD8 T cells, expression of the early activation markers CD25 and CD69 does not affect CD8 T cells secrete IL 2 and IFN-Ah. The same is true in vivo experimental results. 3 co-culture DCreg mDCs and CD8 T cells, IL-10 secretion increased significantly, but blocking the IL-10 did not reverse the DCreg inhibition of antigen-specific CD8 T cells. 4 After stimulation with LPS DCreg the hypersecretion NO. In the mDCs CD8 T DCreg culture system by blocking the supernatant NO DCreg antigen-specific CD8 T cell inhibition reversed, suggesting that NO plays a key role in DCreg of antigen-specific CD8 T cell inhibition. Conclusion (1) LSCs to express a variety of cytokines and chemokines. (2) LSCs make mDCs continue to proliferate and differentiate into immune negative regulation DCreg. (3) LSCs induce DCreg able to inhibit the proliferation of antigen-specific CD8 T cells, but does not affect the activation of CD8 T cells, NO has played a key role in the inhibition. (4) LSCs DCreg induced by in vivo reinfusion in vivo inhibition of antigen-specific CD8 T cell proliferation. The significance of this experiment show that the liver microenvironment to promote mDCs differentiated DCreg, the proliferation of antigen-specific CD8 T cells inhibit this inhibition mainly mediated by NO. The study contributes to a deep understanding of the role of the immune microenvironment of the liver and liver immune cell subsets in the liver tolerance and maintain immune balance.

Suppression of CD8~+T Cells by Regulatory Dendritic Cells

Abstract

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