Regulatory T Cells Through The Secretion Of Intracellular Factors TGF?1 Promote The Expression Of CD133 Glioma Stem Cells

Background and objectivesGlioma is one of human fatal tumor,in the past five years in spite of the treatment is improved,median survival time of glioma is just 14 months after diagnosis^[1],the prognosis is very poor.In the United States and Europe each year about 77000 people are diagnosed with VI grade glioma,and 5-year survival rate less than 3%,for recurrent glioma patients,survival time only a few months.Tumor recurrence is an important mechanism for the existence of glioma stem cells and glioma stem cells with self-renewal,proliferation and differentiation potential,although small,but in the occurrence of tumors,plays an important role in the development,recurrence and metastasis.Recurrence of malignant glioma stem cells with self-renewal potential,one of the important reasons that lead to high mortality rate,in the past ten years,more and more evidence to support such a view,namely the tumor stem cells or starting,responsible for development including angiogenesis,invasion and lead to tumor treatment resistance,and recurrence.Recurrence of the disease may be caused in the treatment of most brain glioma cell death in the process,increase in disease progression or obtain the characteristics of the stem cells.So the glioma stem cells is an important therapeutic targets.Tumor immunotherapy in recent years become the focus in the tumor treatment.Junwei think glioma initial sample cells induced immunosuppression?glioma stem cells?can be thought of^[1].Existing literature on glioma patients'survival time and its closely related to immune status.In recent years the rapid development of tumor immunology by more and more people's attention,the body's immune system by normal cells immune mechanism to identify and eliminate tumor cells,when the body's immune function is low,immune surveillance function to drop,the tumor cells to evade the host immune system attacks began to grow.Tumor microenvironment and Tumor is closely tied to the occurrence and progress of the growth of Tumor cells without Tumor microenvironment,the survival of cancer stem cells is not exceptional also,Tumor microenvironment,including Tumor Associated macrophage?Tumor-Associated Microglia/MacrophagesTAMS,?,source of marrow inhibitory cells?MDSCs?,regulatory CD4+CD25+FOXP3+T cells?regulatory T cell,Treg?,and some related cytokines,chemokines and cell matrix.1995 by Sakaguchi,found the important negative immunity regulating ingredients-regulatory CD4+CD25+T cells?regulatory T cell,Treg?,as a kind of unique immune regulating function cells,the tumor microenvironment plays an important function of immunosuppression,including FOXP3 as indicators of the group of the specificity of the cell phenotype,its expression was found to be necessary and sufficient inhibitory activity.According to reports in the literature,can be divided into CD4+FOXP3+,CD8+FOXP3+,NKTFOXP3+and double negative.At the same time,on the basis of CD45RA expression?an indicator of T cell differentiation?of high and low,and Treg can be divided into CD45RA+FOXP3+,CD45RA FOXP3+?high?,CD45RA FOXP3+?low?,r-Treg,non-Treg,a-Treg,in these three group of cells,the main work is aTreg.Regulatory T cells through the TIM3 CD73,PD,such as surface inhibitory molecule and secretion TGFB intracellular factor,IL-10 play a negative regulating function,etc.Existing literature reported in glioma microenvironment tumor associated macrophages through the secretion of cytokines TGF?1 promote the expression of glioma stem cells^[3].In this experiment,we are still starting from the tumor microenvironment,try to explore regulatory T cells?Treg?cells of glioma stem cells,for later killed in the process of clinical treatment of stem cells,reduce the recurrence rate and improve the survival of patients with glioma cycle to provide a new therapeutic targets.MethodsCollect glioma patients postoperative 2-4 hours fresh tissue and peripheral blood in patients who 6 ml,healthy human peripheral blood of 200 ml,adopt mechanical separation processing organization,and cryopreserved density gradient centrifugation separation of peripheral blood mononuclear cells,rt-pcr detection of glioma tissues CD133 TGF?1 foxp3 gene expression level and to analyze its correlation;Flow cytometry analysis of peripheral blood CD4+FOXP3+Treg,R-Treg,Non-Treg,A-Treg expression level and the three group of TGF?1 cells secrete cell factor;By ELISA-enzyme-linked immunosorbent experiment on test altogether after incubation level of cytokine secretion of TGF?1,Transwell experimental detecting chemokine CCL2 chemotaxis of Treg function.Results1.The expression of the regulatory T cells in peripheral blood is higher than glioma tissues.The expression level of stem cell gene CD133 have a relationship with FOXP3 in glioma tissue.2.The CD133 is a maker of glioma stem cells.3.The CD4+FOXP3+regulatory T cells to promote the expression of CD133glioma stem cells.4.A-Treg regulatory T cells secrete high level TGF?1 promote glioma CD133expression.5.CD133 glioma stem cells secreted chemokine CCL2 recruiting Treg reach the tumor site,Treg cells express CCL2 receptors CCR4.Conclusion1.ATreg is active state regulatory T cells,which secrete the higher levels of TGF?1,promote the expression of glioma stem cells,glioma stem cells secrete CCL2recruiting Treg reach the tumor site,Treg cells surface expressionCCL2 receptor CCR4.2.TGF?1 blockers,CCL2 blockers,CCR4 blockers are likely to be clinical kill glioma stem cells,reduce glioma recurrence,improve the clinical treatment of survival in patients with new targets.