目的针对中国汉族高频HLA-DRB1基因型人群,预测与鉴定幽门螺杆菌(Helicobacter pylori,H.pylori)黏附素蛋白A亚单位(neuraminyllactose-binding hemagglutinin,HpaA)CD4+T细胞表位。方法通过Allele Frequency Net数据库分析中国汉族人群HLA-DRB1高频基因型,利用NetMHCIIpan对HpaA蛋白中能被这些高频基因型递呈的CD4+T细胞表位进行预测;通过PCR-SBT法筛选携带有这些高频基因型的H.pylori感染者,利用重组HpaA抗原刺激感染者外周血单个核细胞体外扩增出HpaA抗原特异性T淋巴细胞后利用合成短肽对所预测表位进行鉴定,通过抗体阻断实验对表位的HLA限制性进行分析,利用DC细胞负载重组HpaA蛋白对表位的自然递呈特征进行分析。结果中国汉族人群中HLA-DRB1高频基因型为DRB1*0901(14.4%)、DRB1*1202(13.3%)和DRB1*1501(10.8%),针对上述3种高频基因型预测出24个HpaA蛋白CD4+T细胞表位。其中,4个表位:HpaA39-53(HLA-DRB1*0901)、HpaA42-56(HLA-DRB1*0901)、HpaA89-103(HLA-DRB1*1202)和HpaA87-101(HLA-DRB1*1501)可有效刺激体外扩增的HpaA特异性T细胞产生高水平IFN-γ,且均能被DC细胞自然递呈。结论鉴定得到的4个HLA-DRB1限制性CD4+T细胞表位有可能成为H.pylori表位疫苗设计的候选抗原。
Objective: To investigate the impact of Qing'e Pill(青娥丸,QEP) on the cancellous bone microstructure and its effect on the level of β-catenin in a mouse model of postmenopausal osteoporosis.Methods: Ninety-six 8-week-old speci?c pathogen free C57 BL/6 mice were randomly divided into 4 groups(24/group): sham,ovariectomised osteoporosis model,oestradiol-treated,and QEP-treated groups.Three months after surgery,the third lumbar vertebra and left femur of the animals were dissected and scanned using micro-computed tomography(micro-CT) to acquire three-dimensional(3 D) parameters of their cancellous bone microstructure.The impact of ovariectomy,the effect of oestradiol and QEP intervention on cancellous bone microstructure,and the expression of β-catenin were evaluated.Results: The oestradioland the QEP-treated groups exhibited a signi?cant increase in the bone volume fraction,trabecular number,trabecular thickness,bone surface to bone volume ratio(BS/BV),and β-catenin expression compared with those of the model group(P<0.05).In contrast,the structure model index,trabecular separation,and BS/BV were signi?cantly decreased compared with those of the ovariectomised osteoporosis model group(P<0.05).No differences were observed in the above parameters between animals of the QEP-and oestradiol-treated groups.Conclusions: The increased β-catenin expression may be the mechanism underlying QEP's improvement of the cancellous bone microstructure in ovariectomised mice.Our ?ndings provide a scienti?c rationale for using QEP as a dietary supplement to prevent bone loss in postmenopausal women.
SHUAI BoZHU RuiYANG Yan-pingSHEN LinXU Xiao-juanMA ChenLU Lin
