ELISPOT与Cultured ELISPOT检测宫颈癌HPV16抗原多肽特异性CTL反应分析  

Analysis of cervical cancer HPV16 of antigen peptides specific CTL reaction by ELISPOT and Cultured ELISPOT

作　　者：马苗苗[1] 常雪梅[1,2] DONG Tao 王若峥[1,2] 

机构地区：[1]新疆医科大学附属肿瘤医院放疗中心,乌鲁木齐830011 [2]新疆维吾尔自治区肿瘤学重点实验室,乌鲁木齐830011 [3]英国牛津大学MRC人类免疫中心

出　　处：《新疆医科大学学报》2017年第7期862-865,870,共5页Journal of Xinjiang Medical University

基　　金：国家自然科学基金(U1603282)

摘　　要：目的比较酶联免疫斑点法(ELISPOT)和Cultured ELISPOT检测宫颈癌HPV16全基因组抗原多肽特异性CTL反应水平差异。方法选择2015年9月-2017年3月新疆医科大学附属肿瘤医院病理明确诊断宫颈鳞状细胞癌32例患者,采用ELISPOT试验(19例,ELISPOT组)和Cultured ELISPOT试验(13例,Cultured ELISPOT组),比较2组HPV16抗原多肽特异性CTL反应频率及强度差异。观察HPV16全基因组抗原多肽E1-1、E1-2、E2、E4、E5、E6、E7、L1及L2刺激后的免疫反应。结果 Cultured ELISPOT组各孔HPV16抗原多肽E1-1、E1-2、E2、E4、E5、E6、E7、L1及L2斑点数高于ELISPOT组。ELISPOT组与Cultured ELISPOT组各肽段间特异性CTL反应频率比较,差异无统计学意义(P>0.05)。Cultured ELISPOT组特异性CTL细胞数显著高于ELISPOT组,2组E1-1、E1-2、E2、E4、E6、E7、L1及L2肽段特异性CTL反应强度比较,差异有统计学意义(P<0.05)。从反应频率及强度观察,L1肽段略高于其他肽段。结论 Cultured ELISPOT检测法与ELISPOT相比有较高的灵敏性,该方法可用于特异性优势肽段筛选。Objetive To compare the differences of cervical cancer HPV16 Genomic Antigen Peptide Specific CTL Response Level by enzyme-linked immunospot assay （ELISPOT） and Cultured ELISPOT. Methods 32 cases selected were newly diagnosed cervical squamous cell carcinoma, and all patients were pathological diagnosis confirmed. The patients were randomly divided into the ELISPOT group （19 cases） and the Cultured ELISPOT group （13 cases） to analyze the frequency and intensity of specific CTL re-sponse between the two groups, and observe specific CTL response of HPV16 genome antigenic polypep-tides of El-1, El-2 , E2 , E4, E5 , E6 , E7 , LI and L2. Results The number of HPV16 antigen peptides of El-1, El-2, E2, E4, E5 , E6 , E7 , LI and L2 were higher in the Cultured ELEPOT group than the ELIS-POT group. There was no significant difference in the frequency of CTL response of peptides between the ELISPOT group and the Cultured ELISPOT group （P 〉0.05）. The number of specific CTL cells in the Cultured ELISPOT group was significantly higher than the ELISPOT group, and the specific CTL re-sponse intensity of El-1, El-2, E2, E4, E6, E7, LI and L2 peptides in the two groups was statistically significant （P 〈0.05）. The LI peptides were a little higher than other peptides from the reactionfrequency and intensity. Conclusion Cultured ELISPOT has a higher sensitivity than ELISPOT, which could be used for specific advantage peptides screening.

关 键 词：宫颈癌 酶联免疫斑点法 HPV16型抗原肽 

分 类 号：R737.33[医药卫生—肿瘤]