Assar Shokrollah, Arababadi Mohammad Kazemi, Mohit Maryam, Ahmadabadi Behzad Nasiri, Pumpens Paul, Khorramdelazad Hossein, Hajghani Masomeh, Assar Sepideh, Araste Majid, Nekhei Zohre, Sendi Hossein, Kennedy Derek
Dept. of Microbiology, Hematology and Immunology, Faculty of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.
Clin Lab. 2012;58(1-2):53-60.
Escape mutations potentially allow viruses to avoid detection and clearance by the host immune system and may represent a mechanism through which infections may persist in some patients. The association of the mutations in the HBcAg gene with Hepatitis B asymptomatic carriers (ASC) has not been studied adequately. The current study was aimed to investigate HBcAg18-27 CTL epitope mutations in ASC patients in the South-Eastern region of Iran.
100 ASC patients were selected for this study and screened for HLA-A2 using flow cytometry. HBV-DNA was extracted from the HLA-A2 positive patients and the HBc gene was amplified using PCR. Direct double sequencing was performed to analyse mutations in the HBc gene of HBV isolates from patients with ASC.
Overall, 25 (25%) of individuals were HLA-A2 positive. Direct double sequencing indicated no mutations in the HBcAg18-27 epitope. However, four mutations within the T helper and three mutations within the B cell epitopes of ASC patients were identified.
The lack of mutations within the HBcAg18-27 epitope suggests that the antigenicity of this region is not altered in HBV isolates of our patients and therefore antigen presentation would occur normally to the patient's immune system through HLA-A2. However, in the course of this study we revealed some novel mutations within the T helper and B cell epitopes that may affect the efficiencies of immune response of ASC patients against these novel HBV epitopes.
逃逸突变可能使病毒逃避宿主免疫系统的检测和清除,这可能是感染在一些患者体内持续存在的一种机制。乙肝核心抗原(HBcAg)基因中的突变与乙肝无症状携带者(ASC)的关联尚未得到充分研究。本研究旨在调查伊朗东南部地区ASC患者中HBcAg18 - 27细胞毒性T淋巴细胞(CTL)表位突变情况。
本研究选取100例ASC患者,采用流式细胞术检测HLA - A2。从HLA - A2阳性患者中提取乙肝病毒脱氧核糖核酸(HBV - DNA),并通过聚合酶链反应(PCR)扩增HBc基因。对ASC患者的HBV分离株的HBc基因进行直接双向测序以分析突变情况。
总体而言,25例(25%)个体为HLA - A2阳性。直接双向测序表明HBcAg18 - 27表位无突变。然而,在ASC患者的辅助性T细胞表位内发现4个突变,在B细胞表位内发现3个突变。
HBcAg18 - 27表位缺乏突变表明该区域的抗原性在我们患者的HBV分离株中未发生改变,因此通过HLA - A2可正常向患者免疫系统呈递抗原。然而,在本研究过程中,我们在辅助性T细胞和B细胞表位内发现了一些新突变,这些突变可能会影响ASC患者针对这些新的HBV表位的免疫反应效率。