WTAP基因DNA甲基化与广西汉族儿童乙肝疫苗低/无应答水平的关联性

The relationship between DNA methylation of WTAP gene and low/nonresponse level of hepatitis B vaccine in Guangxi han children

目的探讨WTAP基因DNA甲基化与广西汉族儿童乙肝疫苗(HepB)应答水平的关联性,以期了解其他可能影响乙肝疫苗应答水平的遗传因素,为新型HepB的研发提供新的科学依据。方法选取在2016年1月至12月间至广西3家医院就诊的8~9月龄且全程接种HepB的263名汉族儿童作为研究对象。对研究对象进行乙肝两对半检测,根据排除和纳入标准筛选符合要求的研究对象纳入本次研究。根据乙肝表面抗体浓度(抗-HBs)将研究对象分成无应答(<10 mIU/mL)、低应答(10 mIU/mL≤抗-HBs<100 mIU/mL)、正常应答(100 mIU/mL≤抗-HBs<1000 mIU/mL)和高应答(≥1000 mIU/mL)组,使用非匹配病例对照研究的方法,将抗-HBs<100 mIU/mL作为无/低应答组,≥100 mIU/mL作为正常/高应答组。采用Panel多重PCR技术、重亚硫酸盐转化技术和通过illumina平台进行高通量测序技术,分析研究WTAP基因38个CpG位点DNA甲基化与HepB免疫应答水平的关系。结果经Mann-Whitney U检验比较无/低应答组和正常/高应答组WTAP基因的DNA甲基化水平后发现,无/低应答组WTAP_2基因43位点DNA甲基化水平低于正常/高应答组DNA甲基化水平(P<0.05)。非条件Logistics回归分析结果显示,WTAP_2基因的43位点DNA甲基化水平和HepB应答水平存在关联性(OR=0.73,95%CI:0.538~0.995),43位点DNA低甲基化水平降低了HepB低/无应答的风险。结论WTAP_2基因的43位点的DNA低甲基化水平可能是广西汉族儿童HepB免疫低/无应答水平的保护因素,可为提高HepB低/无应答人群提高疫苗应答水平提供理论性参考依据。

Objective To explore the relationship between WTAP gene DNA methylation and the response level of HepB vaccine(HepB)in Guangxi Han children,in order to understand other genetic factors that may affect the response level of hepatitis B vaccine,and provide a new scientific basis for the development of new HepB.Methods A total of 263 Han children from 8 to 9 months old who were vaccinated with HepB from January to December 2016 in three hospitals of Guangxi were selected as the research subjects.The study subjects were tested for hepatitis B in 5 basic hepatitis B indicators(two and a half),and the study subjects meeting the requirements were screened for inclusion in this study according to the exclusion and inclusion criteria.According to the concentration of hepatitis B surface antibody(anti-HBs),the subjects were divided into non-response(<10 mIU/mL),low response(10 mIU/mL≤anti-HBs<100 mIU/mL),and normal response(100 mIU/mL≤anti-HBs<1000 mIU/mL)and high response(≥1000 mIU/mL),using the method of unmatched case-control study,with anti-HBs<100 mIU/mL as the non-response group,and≥100mIU/mL as the normal and high response group.Using Panel multiplex PCR technology,bisulfite conversion technology and high-throughput sequencing technology through Illumina platform,the relationship between DNA methylation at 38 CpG sites of WTAP gene and HepB immune response level was analyzed and studied.Results The Mann-Whitney U test was used to compare the DNA methylation levels of the WTAP gene in the nonresponse and low response group and the normal and high-response groups,and the level of DNA methylation at position 43 of WTAP_2 gene in the non-response and low response group was significantly lower than that in the normal and high-response groups(P<0.05).Unconditional logistics regression analysis showed that the level of DNA methylation at position 43 of WTAP_2 gene was correlated with the level of HepB response(OR=0.73,95%CI:0.538-0.995),and the reduced level of DNA methylation at position 43 decreased the risk of low HepB/non-response.Conclusion The DNA hypomethylation level at position 43 of WTAP_2 gene may be a protective factor for the low/non-response level of HepB in Han children in Guangxi,and it can provide a theoretical reference for improving the level of vaccine response in people with low/non-responsive HepB.