不同宫颈病变高危型人乳头状瘤病毒载量对局部调节性T细胞表达的影响

Effects of high risk human papillomavirus loading on the expression of locally regulatory T cells in different cervical lesions

目的比较宫颈局部微环境高危型人乳头状瘤病毒载量(HR-HPV DNA)和CD4+CD25+Foxp3+调节性T细胞(Treg)的同步变化,探讨HR-HPV病毒复制和宫颈病变进展两者同时对Treg细胞的影响。方法将304例HR-HPV持续感染者依宫颈病变的不同分为5组:宫颈上皮内瘤变(CIN)Ⅰ、CINⅡ、CINⅢ、宫颈癌和慢性宫颈炎,采用PCR荧光法检测宫颈分泌物HPV-DNA,应用流式细胞仪CD4+CD25+Foxp3+设门方法对宫颈刷检物中的CD4+CD25+Treg细胞相对计数,对数据资料进行统计学分析。结果(1)宫颈局部Treg细胞表达在不同程度的宫颈病变和不同拷贝数的病毒载量之间比较差异均有显著性(F=24.93,109.86,P<0.05),进一步的两两比较显示,Treg细胞的水平变化在慢性宫颈炎和CINⅠ之间以及低载量(HPV DNA 104~105copies/m L)和中载量(HPV DNA 105~106copies/m L)之间差异无统计学意义(P>0.05),其他组组间比较差异有统计学意义(P<0.05);(2)以Treg细胞水平变化为变量,宫颈病变和病毒载量为因子的交互效应显著(F=3.39,P<0.05),不同的宫颈病变,HR-HPV病毒载量对Treg表达的影响大小不一,总体呈现随着宫颈病变程度加重,HR-HPV病毒拷贝数升高,Treg细胞表达Foxp3+逐步上升的趋势;(3)宫颈癌患者高表达CD4+CD25+Foxp3+Treg细胞,但表达水平波动范围大,数值分布最为分散。结论宫颈局部微环境CD4+CD25+Foxp3+Treg细胞的免疫抑制功能在不同宫颈病变和不同HR-HPV DNA中可能通过双向性调节,影响宫颈病变转归,就整体变化而言,Treg细胞、HR-HPV载量、宫颈病变三者呈相向的进展趋向。

Objective To compare the synchronous changes of high risk human papillomavirus load （ HPV -DNA） and CD4^＋CD25^＋Foxp3^＋ regulatory T cells （Treg） in local microenvironment of cervix, and investigate the ef- fects of HPV virus replication and progression of cervical lesions on Treg cells. Methods 304 cases of HR-HPV infection with cervical lesions were divided into 5 groups, cervical intraepithelial neoplasia （CIN）Ⅰ, CINⅡ, CINⅢ, cervical cancer and chronic cervieitis. The HPV-DNA of cervical secretion was detected by PCR fluorescence, and the relative Treg cells numbers from cervical brush samples were determined by flow cytometry with CD4^＋CD25^＋ Foxp3^＋gating, and the data were statistically analyzed. Results （ 1 ） There was significant difference of cervical Treg cells in different degrees of cervical lesion and different copy numbers by variance comparison （F = 24.93, 109.86, P 〈 0.05 ）, and a further pairwise comparison showed that there was no significant difference of Treg cell between chronic cervicitis and CINI and low load （HPV DNA 10^4 - 10^5 copies/mL） and medium load （HPV DNA 10^5 - 10^6 copies/mL） （P 〉 0.05 ）. There was a significant difference between the other groups （P 〈 0.05 ） ; （2） Treg cells as variable, interaction effect of cervical lesions and viral load factors were significant different （F = 3.39, P 〈 0.05 ）. The effect of different cervical lesions and HR-HPV viral load on the expression of Treg cells was differential. An overall showing with the degree of cervical lesions increased, HR-HPV virus copy number increased, Treg cells expression increased gradually; （3）CD4^CD25^＋Foxp3^＋Treg cells were highly expressed in cervical cancer patients, but the expression level fluctuated widely and the numerical distribution was the most dispersedly. Conclusion The immune suppression function of local CD4^＋CD25^＋Foxp3^＋Treg cells with different cervical lesions and different HR-HPV DNA may bilaterally regulate the prognosis of cervical lesions, as a whole, between Treg cells and HR-HPV load and cervical lesions were the consistent progress trend.