Preliminary proteomic analysis of differential expressed proteins identified by two-dimensional fluorescence difference gel electrophoresis for hepatitis B virus vaccine nonresponders
1. Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China; 2. State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China
Abstract: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.