Detection Of The Intracellular Cytokine Expression And Study Of The Polarization Of T Lymphocyte In Patients With Condyloma Acuminatum

Condyloma acuminatum, the clinical visible manifestations of infection with human papillomavirus(HPV), is the most prevatent sexually transmitted diseases ( STDs) in Europe and Americano In our country,the incidence rate of CA is increasing annually.The HPV also has been causally linked with cervix cancer.At present,there still have no effective immunotherapies targeted at HPV and recurrence occurs frequently. Recurrence of CA is closely associated with the immune system of the host, especially the cell-mediated immune response.Both CD4+and CD8+ T lymphocytes can differentiate into two effector subsets:Thl or Tel and Th2 or Tc2. These two types of cells produce distinct profiles of cytokines and regulate different immune responses. In this study, we are to detect the various intracellular cytokines produced by T cell subsets and to discuss the polarization of Th or Tc lymphocytes in CA patients.The purpose of the study :1. To detect the expression of intracellular cytokines(IL-4 and IFN-Y ) in the peripheral blood T lymphocytes of CA patients and to assessthe polarization of Th 1 ,Th2, Tc1/Tc2.2. To evaluate the variety of the quantity of the different cytokines produced by functional subsets.3. To detect the T lymphocyte surface epitopes and the distribution of Th1/Th2, Tc1/Tc2, then analysis the correlation between them.To probe the difference of two methods and compare the significance of them in assessing the body immune system function.Materials and methods:1. After that the lymphocytes were activated, fixed N permeabilized and stained in vitro, three-color flow cytometry method is used to detect the T cell intrcelluar cytokine ( IL-4 , IFN- r ) expression with fluorochrome conjugated anti-cytokine antibodies, then compared the polarization of Th and Tc subsets between the CA patients and the healthy group.2. Detected the mean fluorescence intensity(MFI) of the 11-4 positive cells or IFN- Y positive cells in Th and Tc subsets by FCM.3. Detected T lymphocyte surface epitope (CD3 > CD4> CDS) combined with simultaneous analysis of the distribution of Thl/Tl^ Tcl/Tc2 subsets,then discussed the correlation between them .Resultes:1 . The percentage of Thl and Tel cells of CA patients are significantly lower than that of heathy control(P<0.01 and P<0.05) .but the the percentage of Th2, Tc2 cells are increased. The mean Thl/Th2 and Tcl/Tc2 ratio of CA patients also decreased significantly (P<0.01)2. Compared with the control group,the Thl cells has lower MFI (P<0.05 ) ,but in Th2 and Tc2 cells ,the MFI is higher .3. Analysis the correlation between the surface antigen detection andthe polarization of T cells shows that the CD4 is negative correlated with Th2,but the ratio of CD4/CD8 is positive correlated with the ratio of Thl/Th2,ands the ratio of Thl/Th2 and Tcl/Tc2 also shows positive correlationship. Conclusion: 1. The CA patients get polarization bias in the peripheral blood Tlymphocytes,either CD4+orCD8+.2. There exist the distinction of not only the percentage of Th and Tc cells, but also the production cytokines ability of T lymphocyte.3. To evaluate the immune function, detecting the functional subsets is more effective than to classifying the surface epitope.