Influence Of Vaginal Flora, T Cell Subsets And Inflammatory Cytokines For Human Papillomaviral Infection

Objectives:To research vaginal flora, T cell subsets and inflammatory factors of cervixin different grade cervical lesions which associated with HPV infection, andstudy the relationship of vaginal flora and immune function in cervix，thenanalysis synthetically their impact on HPV infection and cervical cancer. Hopeto provide an experimental basis for improving cervical microenvironment, andprovide a reference opinion for the intervention and clear of persistent infectionof HPV, and provide new ideas for the prevention and treatment of cervicalcancer.Methods:The patients in this study were those who visited in Gynecologic Oncologybecause of vaginal pruritus or suspected cervical lesions from March2012toOctober2013of Guangxi Medical University Cancer Hospital, then selected180cases as experimental group who had infected with HPV and cervical tissuewere confirmed for CIN or cancer by pathology. Those including134cases inCIN group (CINⅠ grade35cases, CINⅡ grade42cases, CIN Ⅲgrade57cases)and46cases in cervical cancer group, then60cases with HPV-negative and cervical tissue were confirmed normal as the control group. All patients weretaken vaginal secretions, cervical secretions, cervical exfoliated cells andcervical tissues in their first outpatient visit. The types of HPV infection weredetected using a gene-chip technique; PH value were detected using special teststrips; trichomonad were observed by suspension wet filmmethod; mycete, cluecells, neisseria gonorrhoeae, lactobacillus were observed by Gram staining;mycoplasma and chlamydia were detected by special test kit; CD4+, CD8+Tcells were detected by flow cytometry and immunohistochemistry; inflammatorycytokines IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10were detected by enzymelinked immunosorbent assay (ELISA).Results:1. Single HPV genotype infection were showing a increasing trend inCINⅠ, CIN Ⅱ, CINⅢ and cervical cancer group, but multiple infections weredecreasing (P <0.05). Eighteen kinds of HPV genotypes were detected. Thedetection rate of high-risk HPV were much higher than low-risk, and the topthree is HPV16,52and58. HPV16infection were showing a increasing trend indifferent pathological (P <0.05).2. The positive of bacterial vaginosis, ureaplasma urealyticum, chlamydiaand negative of lactobacillus were showing a increasing trend in control group,CIN Ⅰ, CIN Ⅱ, CINⅢ and cervical cancer groups (P <0.05); In contrast, thepositive of trichomonad, mold and mycoplasma hominis was not different (P>0.05); neisseria gonorrhoeae were not detected in all samples, and we can notanalyze the relationships of neisseria gonorrhoeae infection and cervical lesions.3. The percentage of CD4+T in cervical secretions were showing adecreasing trend in control group, CINⅠ, CINⅡ, CINⅢ and cervical cancergroup (P <0.05), but the percentage of CD8+T was no difference (P>0.05), CD4+/CD8+were also decreasing (P <0.05). With the increasing of thepathological grade, the expression of CD4were showing a decreasing trend andCD8were increasing in cervical tissue, and the percentage of CD4+/CD8+<1were also showing a increasing trend (P <0.05).4. The expression of IFN-γ, IL-2were showing a decreasing trend and IL-4was increasing in cervical secretions in the control group, CINⅠ, CINⅡ,CIN Ⅲ and cervical cancer group, and IFN-γ/IL-4, IL-2/IL-4were alsoreducing (P <0.05); but TNF-α, IL-6, IL-10were no difference (P>0.05).5. The positive of bacterial vaginosis, mycoplasma hominis and chlamydiawere all independent risk factors for the decreacing of IL-2(P＜0.05). Thepositive of bacterial vaginosis, ureaplasma urealyticum and chlamydia were allindependent risk factors for the increacing of IL-4(P＜0.05).6. The results of HPV as the dependent variable, all other factors as theindependent variables were analyzed with single factor logistic regressionshowed that difference of lactobacillus, IFN-γ, IL-2and IL-4were significant inHPV-positive and HPV-negative groups (P <0.05). In the multiple factorslogistic regression analysis, the finally result was that IFN-γ, IL-2, IL-4relatedwith HPV infection observably (P <0.01).Conclusions:1. The development of cervical lesions did not depend on the types of HPV,but for the pathogenicity. A single high-risk HPV infection is easily to increasethe occurrence of cervical cancer, and multiple infections not increased the riskof cervical cancer.2. The increasing infection of bacterial vaginosis, ureaplasma urealyticum,chlamydia and the decreasing infection of lactobacillus were closely related tothe occurrence and development of cervical cancer. 3. The increasing infection of bacterial vaginosis, ureaplasma urealyticum,mycoplasma hominis and chlamydia were closely related to the decreacing oflocal immunity in cervix.4. With the increasing of the pathological grade, CD4+/CD8+and Th1/Th2in cervix were reducing gradually. Above all indicated that decreasingimmune function in cervix can promote the occurrence and development ofcervical cancer.5. The content of IFN-γ, IL-2were negatively correlated with HPVinfection, but the negative of lactobacillus and content of IL-2were positivelycorrelated with HPV infection. The inflammatory cytokines decreacing of IFN-γ,IL-2and the increasing of IL-4were all independent risk factors for HPV.