Abstract: Background　Persistent infection of high-risk human papillomavirus （HR-HPV） is necessary but not sufficient for the occurrence of cervical cancer.Multiple changes in genome，microenvironment and immune system，and inflammation contribute to carcinogenesis.Therefore，the biomarkers of HR-HPV infection detected simply and noninvasively that can accurately and quantitatively reflect the changes in biological behavior of cervical cancer，become a focus of clinical research.Objective　To examine the changes in HR-HPV load and regulatory T Cells （Treg） in the same microenvironment of cervical lesions，and based on this，to discuss their interactions and expressions in the development of cervical lesions.Methods　We enrolled 342 cervical lesions patients with persistent HR-HPV infection from First Affiliated Hospital of Henan Polytechnic University during January 2012 to July 2015，including 55 with chronic cervicitis，73 with cervical intraepithelial neoplasia（CIN）Ⅰ，71 with CINⅡ，72 with CINⅢ and 71 with squamous carcinoma of the cervix （SCC）．QF-PCR was used to detect the HR-HPV load.According to the level of HR-HPV load （copy number of HPV DNA），we divided the participants into 4 groups：low load （HPV DNA 104-105 copies/ml，n=69），medium load （HPV DNA>105-106 copies/ml，n=90），high load （HPV DNA>106-107 copies/ml，n=106）and very high load （HPV DNA>107 copies/ml，n=77）．Flow cytometry was used to determine the number of Treg from cervical samples collected using a cytology brush.Results　There were significant differences in the number of Treg among different load groups （P<0.05）．The number of Treg increased with the growth of HR-HPV load，namely，low load group had less number of Treg than other 3 groups（P<0.05），so did the medium load group when compared with other 2 high groups （P<0.05），and as did the high load group when compared with very high group （P<0.05）．The number of Treg differed significantly in patients with different cervical lesions （P<0.05）．Chronic cervicitis patients had less number of Treg than other patients，so did CINⅠpatients when compared with CINⅡ，CINⅢ and SCC patients，and so did CINⅡpatients when compared with CINⅢ and SCC patients，and as did the CINⅢ patients when compared with SCC patients（P<0.05）．The number of HR-HPV load differed significantly in patients with different cervical lesions（P<0.05）．Chronic cervicitis and CINⅠpatients had less HR-HPV load than other patients，so did CINⅡ and CIN Ⅲ patients when compared with SCC patients （P<0.05）．Clustered boxplot diagram showed that chronic cervicitis and SCC patients were found with a total of 3 slightly abnormal values，but other patients without；the distribution of HR-HPV load was dispersed in SCC patients，but comparatively concentrated in other cervical lesion patients.The distribution of cervical lesions in different HR-HPV load groups was significantly different （P<0.05）．Correlation analysis showed that HR-HPV load was positively correlated with the severity of cervical lesions and the number of Treg （r=0.631，0.683，P<0.05），and there was a positive correlation between the severity of cervical lesions and the number of Treg （r=0.863，P<0.05）．Conclusion　The level of HR-HPV load is not always positively correlated with the severity of cervical lesions.The roles of Treg in the development of different cervical lesions differ by their expressions in the local cervical microenvironment.There are different pathogenicities，which causes immune imbalance，a key link in the development of cervical cancer.

Key words: Uterine cervical diseases；Uterine cervical neoplasms；Cervical intraepithelial neoplasia；Papillomavirus；T-lymphocytes, regulatory

摘要： 背景　高危型人乳头瘤病毒（HR-HPV）持续性感染是宫颈癌发生的必要非充分条件，基因组、微环境、炎症和免疫体系的多环节变化促发了癌变的发生，因此，基于HR-HPV感染并能方便、无创、准确量化地反映宫颈癌生物学行为变化的标志物是临床研究的重点。目的　比较不同宫颈病变患者HR-HPV载量和调节性T淋巴细胞（Treg）的变化，探讨两者在不同宫颈病变的相互作用和差异表达。方法　选取2012年1月—2015年7月河南理工大学第一附属医院收治的HR-HPV持续性感染的不同宫颈病变患者342例，其中慢性宫颈炎55例、宫颈上皮内瘤变（CIN）Ⅰ级73例、CINⅡ级71例、CINⅢ级72例、宫颈癌71例。采用聚合酶链式反应（PCR）荧光法检测HR-HPV载量，根据HR-HPV载量拷贝数分为4组：低载量组（104~105 copies/ml）69例、中载量组（>105~106 copies/ml）90例、高载量组（>106~107 copies/ml）106例、超高载量组（>107 copies/ml）77例，应用流式细胞仪检测宫颈刷取物Treg。结果　不同载量组患者Treg比较，差异有统计学意义（P<0.05）；其中中载量组、高载量组、超高载量组患者Treg较低载量组升高，高载量组、超高载量组患者Treg较中载量组升高，超高载量组患者Treg较高载量组升高（P<0.05）。不同宫颈病变患者Treg、HR-HPV载量比较，差异有统计学意义（P<0.05）；其中CINⅠ级患者Treg较慢性宫颈炎升高，CINⅡ级、CINⅢ级、宫颈癌患者Treg、HR-HPV载量较慢性宫颈炎、CINⅠ级升高，CINⅢ级患者Treg较CINⅡ级升高，宫颈癌患者Treg、HR-HPV载量较CINⅡ级、CINⅢ级升高（P<0.05）。复式箱型图示，在慢性宫颈炎和宫颈癌共有3个温和的异常值，其余病变组无异常值，宫颈癌HR-HPV载量分布分散，低宫颈病变HR-HPV载量分布较为集中。不同HR-HPV载量组宫颈病变分布比较，差异有统计学意义（P<0.05）。相关分析显示，HR-HPV载量与宫颈病变、Treg呈正相关（r=0.631、0.683，P<0.05），宫颈病变与Treg呈正相关（r=0.863，P<0.05）。结论　HR-HPV载量与宫颈病变的严重程度不总是同步的，宫颈局部微环境Treg的不同表达，在宫颈病变程度中发挥的作用也不相同，存在致病差异性，由此造成的免疫失衡是宫颈癌变的关键环节。

关键词: 宫颈疾病；宫颈肿瘤；宫颈上皮内瘤变；人乳头瘤病毒；T淋巴细胞, 调节性