Qin Xinmin, Guo Yonghong, Du Haoqi, Zhong Yaogang, Zhang Jiaxu, Li Xuetian, Yu Hanjie, Zhang Zhiwei, Jia Zhansheng, Li Zheng
Laboratory for Functional Glycomics, College of Life Sciences, Northwest UniversityXi'an, China.
Department of Infectious Diseases, Second Affiliated Hospital of Xi'an Jiaotong UniversityXi'an, China.
Front Physiol. 2017 Aug 21;8:596. doi: 10.3389/fphys.2017.00596. eCollection 2017.
Chronic infection with HBV (CHB) or HCV (CHC) is the most common chronic viral hepatitis that can lead to cirrhosis and hepatocellular carcinoma in humans, their infections have distinct pathogenic processes, however, little is known about the difference of glycoprotein glycopatterns in serum between hepatitis B virus (HBV)- and hepatitis C virus (HCV)-infected patients. A method combining the lectin microarrays, letin-mediated affinity capture glycoproteins, and MALDI-TOF/TOF-MS was employed to analyze serum protein glycopatterns and identify the glycan structures from patients with CHB ( = 54) or CHC( = 47), and healthy volunteers (HV, = 35). Lectin blotting was further utilized to validate and assess the expression levels of their serum glycopatterns. Finally, the differences of the glycoprotein glycopatterns were systematically compared between CHB and CHC patients. As a result, there were 11 lectins (e.g., HHL, GSL-II, and EEL) exhibited significantly increased expression levels, and three lectins (LCA, VVA, and ACA) exhibited significantly decreased expression levels of serum protein glycopatterns only in the CHB patients. However, DBA exhibited significantly decreased expression levels, and two lectins (WGA and SNA) exhibited significantly increased expression levels of serum glycopatterns only in the CHC patients. Furthermore, LEL and MAL-I showed a coincidentally increasing trend in both CHC and CHB patients compared with the HV. The individual analysis demonstrated that eight lectins (MPL, GSL-I, PTL-II, UEA-I, WGA, LEL, VVA, and MAL-I) exhibited a high degree of consistency with the pooled serum samples of HV, CHB, and CHC patients. Besides, a complex-type glycans binder PHA-E+L exhibited significantly decreased NFIs in the CHB compared with HV and CHC subjects ( < 0.01). The MALDI-TOF/TOF-MS results of -linked glycans from the serum glycoproteins isolated by PHA-E+L-magnetic particle conjugates showed that there was an overlap of 23 -glycan peaks (e.g., m/z 1419.743, 1663.734, and 1743.581) between CHB, and CHC patients, 5 glycan peaks (e.g., m/z 1850.878, 1866.661, and 2037.750) were presented in virus-infected hepatitis patients compared with HV, 3 glycan peaks (1460.659, 2069.740, and 2174.772) were observed only in CHC patients. Our data provide useful information to find new biomarkers for distinguishing CHB and CHC patients based on the precision alteration of their serum glycopatterns.
慢性乙型肝炎病毒(HBV)或丙型肝炎病毒(HCV)感染是最常见的慢性病毒性肝炎,可导致人类肝硬化和肝细胞癌,然而它们的感染具有不同的致病过程,关于HBV感染患者和HCV感染患者血清中糖蛋白糖型的差异知之甚少。采用凝集素微阵列、凝集素介导的亲和捕获糖蛋白和基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF-MS)相结合的方法,分析了54例慢性乙型肝炎(CHB)患者、47例慢性丙型肝炎(CHC)患者和35例健康志愿者(HV)的血清蛋白糖型,并鉴定聚糖结构。进一步利用凝集素印迹法验证和评估其血清糖型的表达水平。最后,系统比较了CHB和CHC患者糖蛋白糖型的差异。结果显示,11种凝集素(如HHL、GSL-II和EEL)的血清蛋白糖型表达水平仅在CHB患者中显著升高,3种凝集素(LCA、VVA和ACA)的表达水平显著降低。然而,DBA的表达水平仅在CHC患者中显著降低,2种凝集素(WGA和SNA)的血清糖型表达水平显著升高。此外,与HV相比,LEL和MAL-I在CHC和CHB患者中均呈现出一致的升高趋势。个体分析表明,8种凝集素(MPL、GSL-I、PTL-II、UEA-I、WGA、LEL、VVA和MAL-I)与HV、CHB和CHC患者的混合血清样本具有高度一致性。此外,与HV和CHC受试者相比,复合型聚糖结合剂PHA-E+L在CHB患者中的归一化荧光强度显著降低(P<0.01)。PHA-E+L磁珠偶联物分离的血清糖蛋白中N-连接聚糖的MALDI-TOF/TOF-MS结果显示,CHB和CHC患者之间有23个N-聚糖峰(如m/z 1419.743、1663.734和1743.581)重叠,与HV相比,病毒感染的肝炎患者中有5个聚糖峰(如m/z 1850.878和2037.750),仅在CHC患者中观察到3个聚糖峰(1460.659、2069.740和2174.772)。我们的数据为基于CHB和CHC患者血清糖型的精确改变寻找区分它们的新生物标志物提供了有用信息。