Study Of Regulatory T Cells And Immune State In Hepatocellular Carcinoma Patients

1,Correlation of two methods of detecting peripheral CD4+CD25hi-intTreg in Hepatocellular Carcinoma patients and its clinical significanceAIM:Based on the method of detecting CD4+CD25hi-intCD127lowTreg, we hope to establish the method of Foxp3+Treg and explore correlation of two methods of detecting peripheral CD4+CD25hi-intTreg. We detect the frequency of peripheral Treg in Hepatocellular Carcinoma(HCC) patients, in order to lay the foundation for further study.METHODS:Respectively by flow cytometry using CD127low and Foxp3+ markers to detect peripheral Treg level in 31 cases HCC patients and CD4+CD25hi-intFoxp3+Treg level in 32 controls.RESULTS:We successfully establish the method of detecting nucleus transcription factor Foxp3. We use Foxp3+ and CD127low two methods of detecting peripheral Treg level in HCC patients and values are respectively(10.33±4.47)% and(10.51±4.78)%. These two values are positively correlation. Pearson correlation coefficient is 0.889, P<0.01,the correlation was statistical significance.Curve fitting shows power model is the optimal model and according to the power model we establish the regression equation Y=1.236x(x0.899)(Y is Foxp3+Treg,x is CD127lowTreg).The proportion of peripheral CD4+CD25hi-intFoxp3+Treg accounted for CD4+T cells in HCC patients and healthy controls is respectively(10.33±4.47)% and (7.34±1.76)%, P<0.01,the difference was statistically significant.CONCLUSION:We successfully establish the method of detecting nucleus factor Foxp3,based on the previous method of detecting CD4+CD25hi-intCD127lowTreg. These two values are positively correlation. We can use power model to conversion between the two values.Our study shows the frequency of peripheral Treg in HCC patients is increased. 2,Analysis of peripheral Treg and immune response state in CHB-related HCCAIM:To explore the frequency of peripheral CD4+CD25hi-intFoxp3+Treg and the change of peripheral immune state in CHB-related HCC.METHODS:Using flow cytometry to detect the frequency of peripheral CD4+ CD25hi-intTreg in 46 cases of CHB-related HCC,21 cases of CHB-related LC and 32 controls. At the same time, we detect peripheralγδT cells (CD3+CD4-CD8-),helper T cells(CD3+CD4+CD8-),cytotoxic T cells(CD3+CD4-CD8+),total T cells (CD3+CD19-),nature killer T cells(CD3+CD16+CD56+),nature killer cells(CD3-CD16+CD56+),leukocyte common antigen (CD45+) in HCC patients,LC patients and 7 controls.RESULTS:The proportion of peripheral CD4+CD25hi-intFoxp3+Treg accounted for CD4+T cells in HCC patients,LC patients and healthy controls is (9.21±2.07)%,(8.13±0.69)% and (7.34±1.76)%, respectively, P<0.05(P=0.00),the difference was statistically significant. We analysis peripheral immune state and our study show that the value of NKT cells is respectively (7.09±3.24)%,(7.56±4.70)% and (11.93±8.04)% in LC patients, HCC patients and control;the value of NK cells is respectively (30.13±8.58)%,(10.46±8.06)% and (11.25±8.53)% in LC patients, HCC patients and control; the value of CD45+ cells is respectively(19.82±4.46)%, (10.65±4.59)% and (10.79±7.28)% in LC patients,HCC patients and control;the value of CD4+T cells is respectively (27.56±3.36)%,(35.06±7.72)% and (32.88±8.41)% in LC patients, HCC patients and control;the value of CD8+T cells is respectively (25.01±8.47)%,(13.63±7.04)% and (20.19±9.94)% in LC patients, HCC patients and control;the ratio of CD4+/CD8+ is respectively(1.22±0.45)%, (2.98±1.23)% and(1.86±0.77)% in LC patients, HCC patients and control:the difference was statistically significant, P<0.05.Other two immune indexesγδT cells and total T cells were no significant difference in three group.CONCLUSION:The proportion of peripheral CD4+ CD25hi-int Foxp3+ Treg accounted for CD4+T cells in HCC patients was significantly increased. There are complex immune disorders in HCC patients:NKT cells are increased, CD45+cells and NK cells are decreased; with liver cirrhosis as the control, CD4+T cells are not change, CD8+T cells are increased and CD4+/CD8+ ratio is decreased in peripheral blood in HCC patients;while with healthy as the control, CD4+T cells are increased, CD8+T cells are decreased but are not statistically significant and CD4+/CD8+ ratio is increased in peripheral blood in HCC patients, which is different from the present majority of study and provide us with further research ideas.