| Objective:This experiment mainly observed the effect of Fuzheng decoction on CD34 Cell source the dendritic cells of acute acute T lymphocytic leukemia-Complete Remission Complete Remission(T-ALL-CR),patients,and observed the killing effect of T cells on CEM cells by activating T cell+CEM cells,Effect of Traditional Chinese medicine Fuzheng decoction on DC immune function in patients with acute T-lymphocyte leukemia during complete remission.Methods:1.Use Fuzheng decoction to dispel poison with high,medium and the New Zealand White Rabbit was irrigated with low dosage for 3 days in the early,middle and late period,and the normal control group,was used to prepare the serum and normal rabbit serum of the blood-containing rabbit in the sterile condition after the last irrigation of the stomach 2h.2.In sterile condition,bone marrow aspiration of T-ALL-CR patients was used to obtain bone marrow mononuclear cells by FICOLL density gradient centrifugation,and CD34+cells were obtained by the method of immune magnetic beads separation,and the collected CD34+cells were added to cytokines such as stem cell growth factor,platelet-producing element(thrombopoietin,TPO),Interleukin-3,FMS-like tyrosine kinase-3 together 9 days to promote the proliferation of CD34+ cells.3.Using automatic cell counter to adjust the number of cultured CD34 cells to the high,medium and low doses of Chinese herbal medicine-containing rabbit serum prepared in the first step after 1×106/ml,this process requires cytokines such as mononuclear-macrophage colony stimulating factor Interferon-?(IFN-?),Tumor necrosis factor-a(TNF-?),Granulocyte Macrophage Colony Stimulating Factor(GM-CSF),Interleukin-4(IL-4),common culture induction DC,The experiment can be divided into the high dosage+cytokine Group(FQG+XB)of Fuzheng decoction and the dosage+cytokine Group(FQZ+XB),Fuzheng decoction,low dose+cytokine Group(FQD+XB),cytokines Group(XB),Normal rabbit Serum Group(ZC)were replaced with the other half of the day to replace the culture fluid,The morphological and number of cultured cells were observed by inverted microscope,and the activity of cells was observed by cell counting in the cell culture process.4.After collecting and culturing cells for 9 days,the expression of DCs surface specific antigen CDla\CD80\CD83\CD86and HLA-DR was detected by Flow cytometry(FCM).5.T cells collected from normal and T-All-CR patients were separated and purified from peripheral blood,and the DCs with CD34+cell source was 10:1 proportional to 24-hole culture plate cultured for 4-5 days,and the excitation T cells were compared with CEM cells in different target ratio(10:1,20:1,40:1).The specific killing effect of T cells after DC excitation in five groups was detected by MTT assay in 96-hole plate culture 24h.Results:1.By inverted microscope,it was found that high both medium and low doses of Chinese herbal medicine-containing rabbit serum+cytokines Group and cytokine group can induce the growth of DC with typical cytological morphological features,and the total number of DC cells cultured by Chinese herbal medicine-containing rabbit serum+cytokine Group is more typical than that of simple cytokines group,However,there was no significant difference in cell morphology and number between the serum and cytokines in different doses of Chinese herbal medicine,and the growth of typical DC was not observed in normal rabbit serum group.2.DCs Immunophenotype:The mononuclear cells isolated from the bone marrow of T-ALL-CR patients can induce the DC-specific immunophenotype in the serum culture of Fuzheng decoction-containing rabbits.In the later stage of induction,the detection of DCs surface specific antigen CDla\CD80\CD83\CD86 and HLA-DR were expressed in five groups.The overall trend of the experimental data was observed,and the expression rates of 5 surface specific antigens in different doses of rabbit serum+cytokine Group and cytokine group were statistically significant compared with those of normal rabbits(p<0.05).3.DCs stimulates the killing rateof peripheral blood T cells to CEM cells in normal persons and patients.The killing rateof high,medium and low doses of medicated rabbit serum+cytokines group which stimulates normal person and T-ALL-CR patients to CEM cell is higher than cytokine group and normal rabbit serum group at the time of comparing the same target ratio.This is statistically significant(P<0.05).There is no statisticsdifference betweenthe killing rate which is stipulated by the different dose of medicated rabbit serum+cytokines group with the same target ratio of normal person T against CEM cells and the killing rate to CEM cells by stimulating T-ALL-CR patient T cell(P>0.05).When different target ratio is comparedNormal rabbit serum group,medicated serum with different concentration and dose+cytokines group stipulates the killing rate of T-ALL-CR patient T cell against CEM cell,the effective target ratio was 40:1,there was significant difference between 10:1and 20:1(P<0.05).When the same target ratio was compared,there was no statistics difference in killing rate between the normal and the patients in the same group(P>0.05).Conclusion:1.Fuzheng Qudu decoction can promote the differentiation and maturation of DCs derived from CD34 cells in patients with T-ALL-CR.2.The DCs induced by Fuzheng decoction containing drugs in rabbit serum can stimulate the killing effect of T cells of peripheral blood of patients and normal people on CEM cells,which is beneficial to the function of DCs.3.The T cells in peripheral blood of T-ALL-CR patients have normal immune function. |
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