Study Of The Levels Of T Regulatory Cells And Suppressor Cytokines In Genital Chlamydial Trachomatis Infection Patients

PartⅠ:Establishment of a non-invasive diagnositic technic which can detect genital Chlamydial trachomatis infectionObjective:To establish a non-invasive Chlamdial trachomatis diagnostic technic by comparing the sensitivity and specificity of Clearview test,PCR and SAT technic.Methods:Using Clearview test,PCR and SAT technic to detect Chlamydial trachomatis of 762 patients who visted the STD clinic of Huashan hospital for the first time.Two or more than two tests positive were regarded as ture positive according to the "expanded golden criterion".The sensitivity and specificity of Clearview test,PCR and SAT technic were compared.Results:84 patients were diagnosed as true positive which include of 52 males and 32 females.The sensitivity and specificity of SAT is 98.81%and 99.26% respectively.The sensitivity and specificity of PCR is 98.81%and 99.56% respectively.The sensitivity and specificity of Clearview is 54.76%and 94.10% respectively.There have no significant difference in specificity among the three tests. There have no significant difference in sensitivity between SAT and PCR,while the sensitivity of Clearview test is significantly lower than SAT and PCR.Conclusions:SAT technic is a non-invasive diagnositic test with both high specificity and high sensitivity,and can be used in genital Chlamydial trachomatis dectction.PartⅡ:Determination of CD4~+CD25~+Foxp3~+Treg cells and suppressor eytokines in the peripheral blood of genital chalamyidal trachomatis infecion patientsObjective:To investigate the amount of CD4~+CD25~+Foxp3~+ Treg regulatory T cells in peripheral blood of patients infected with genital chalamyidal trachomatis.Methods:The expression of transcription factor Foxp3 was determined by three-color flow cytometry in peripheral CD4~+CD25~+T cells from 40 patients with genital CT infection and 20 normal controls.Highly purified CD4~+CD25~+T cells were isolated ex vivo from peripheral blood by immunomagnetic beads,and the production of intracellular suppressor cytokines such as interleukin-10(IL-10) and transforming growth factor-β(TGF-β) was tested by flow cytometry.Results:1)The proportion of CD4~+CD25~+Foxp3~+Treg cells was 5.82±0.91% on CD4~+T cells in no inflammation present patients group.The inflammation present patients group was 5.84±0.87%.The normal control group was 5.65±0.78%.The proportion of CD4~+CD25~+Foxp3~+Treg cells have no signifcant difference in the three group(P＞0.05). 2)The proportion of CD4~+CD25~+IL-10~+T cells and CD4~+CD25~+TGF-β~+T cells in no inflammation present patients group was 0.47±0.21%and 0.114+0.05% respcectively.The inflammation present patients group was 0.41±0.17%and 0.10±0.04%respectively.The normal control group was 0.39±0.17%and 0.09±0.04%.The proportion of IL-10 and TGF-βnot had change in the three group (P＞0.05).3)Compared with the control group and inflammation present patients group, the secretion of intracellular cytokines such as IL-10 and TGF-βwas significantly greater in peripheral CD4~+CD25~+T cells of no inflammation present patients group after PMA and ionomycin stimulation(P＜0.05).Conclusions:The amount of CD4~+CD25~+Foxp3~+ Treg regulatory T cells in peripheral blood of patients infected with genital chalamyidal trachomatis has not changed.The suppressor cytokines of CD4~+CD25~+T cells got potentiation in no inflammation present patients group.PartⅢ:Detecting CD25 and Foxp3 in cervical mucosa of genital tract Chlamydial trachomatis infection patientsObjective:To study the CD25 and Foxp3 level in cervix mucous membrane of genital tract Chlamydial Trachomatis infection patients. Methods:The level of CD25 and Foxp3 in cervix mucous membrane were determined by immunofluorescence from 10 patients with no inflammation,10 patients with inflammation present and 10 patients normal controls.Results.The cervix mucous membrane had CD25 and Foxp3 positive cells genital tract after Chlamydial trachomatis infection.The cervix mucous membrane does not appeared CD25 and Foxp3 positive cells in normal controls groups.In no inflammation groups,the number and fluorescence intensity of CD25 and Foxp3 positive cells had higher than the inflammation groups.Conclusions:After genital tract Chlamydial trachomatis infection,peripheral or lymphatic tissue CD25 and Foxp3 positive cells could extension per saltam for local mucous membrance,and be activated.In no inflammation groups,the number and function of CD25 and Foxp3 positvie cells had better than the inflammation groups.PartⅣ:Detecting the suppressor cytokines in cervical mucosa of genital tract Chlamydial trachomatis infection patients Objective:To study the mRNA level of suppressor cytokines in the cervix mucous of genital tract Chlamydial Trachomatis(CT)infection.Methods:The content of IL-10,TGF-β,PD-1,CTLA-4 and GITR in cervix mucous membrane were determined by fluorescent quantitation reverse transcription and polymerase chain reaction from 10 patients with no inflammation,10 patients with inflammation present and 10 normal controls.Results:The content of IL-10,TGF-β,PD-1,CTLA-4 and GITR had significant deviation in three group(P＞0.05).The content of cell factor was lowest in normal control groups.The inflammation groups had higher than the normal control groups,but no inflammation groups had highest content of cell factor.Conclusions:After genital tract infection from CT,peripheral CD4~+CD25~+Foxp3~+ regulatory T cells could extension per saltam for local mucous membrance,and activation,secrete cell factor.The CD4~+CD25~+Foxp3~+ regulatory T cells had proper activation in patients of inflammation.But the CD4~+CD25~+Foxp3~+ regulatory T cells overact activation in patients of no inflammation.The level of suppressor cell factor overact increase.