以往研究显示，人群中约有10%无法通过接种乙型肝炎病毒（HBV）疫苗而产生保护性水平的应答反应。为研究HBV疫苗免疫后不应答发生的机制，本研究对108例接受正常免疫程序（大连汉信产HBV疫苗，10 μg/支，共接种3针）后表达不同应答水平的人群进行血浆蛋白质组差异分析，将其分为无应答（抗体效价检测结果为阴性）组和高应答（抗体效价约1 000 mIU/ml）组。用多重亲和去除系统（MARS）亲和柱去除14种高丰度蛋白质后，以荧光差异双向电泳（2D-DIGE）与基质辅助激光解析电离串联飞行时间质谱（MALDI-TOF-MS）相结合的方法，鉴定获得11种差异表达蛋白质。在这些差异表达蛋白质中，α1微球蛋白、激肽原1的表达在无应答组人群中较高应答组上调，而α1抗胰凝乳蛋白酶、维生素D结合蛋白、Afamin、抗凝血酶Ⅲ和玻连蛋白表达下调。其中，α1抗凝乳蛋白酶、激肽原1和α1微球蛋白的差异性表达进一步得到酶联免疫吸附试验（ELISA）或蛋白免疫印迹法验证。以上蛋白质的表达状况与HBV疫苗免疫应答状况相关，可能成为检测HBV疫苗免疫应答反应水平的分子标记，为进一步研究免疫不应答机制奠定基础。

There is around 10% people who can’t produce sufficient antibody after hepatitis B virus (HBV) vaccine injection. In order to find the mechanism of HBV vaccine non-responsiveness, 108 volunteers were unified injected HBV vaccine (10 μg/shot) three times. They were divided into non-responders with negative HBV surface antibody (HBsAb) titer and high responders with HBsAb titer about 1 000 mIU/ml. The plasma proteome were analyzed by two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) technology after depletion of 14 high abundant proteins by multiple affinity removal system (MARS) column. 11 differential expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Among these proteins, α1-microglobulin and kininogen-1 was up-regulated in nonresponders compared to high responders, while α1-antichymotrypsin, vitamin D binding protein, Afamin, antithrombin Ⅲ and vitronectin were down-regulated. After extensive validation by Western blotting and enzyme-linked immunosorbent assay (ELISA), the differential expression level of α1-antichymotrypsin, kininogen and α1-microglobulin were confirmed. These results suggest that those proteins are associated with the responsive state of HBV vaccine, thus may serve as potential biomarkers for assessing the immune response among diverse HBV vaccine responders and shed light on the mechanism of immunologic unresponsiveness.