Peng Xiao-mou, Xie Dong-ying, Gu Lin, Huang Yang-su, Gao Zhi-liang, Yao Ji-lu
Department of Infectious Diseases, Third Affiliated Hospital, Zhongshan University, Guangzhou 510630, China.
Zhonghua Yi Xue Za Zhi. 2003 Feb 10;83(3):232-6.
To study the effect of exogenous epitope of helper T lymphocyte (HTL) on humoral immunity of HBV S gene DNA immunity.
Two universal HTL epitopes, amino acid residue (aa) 830-843 of the tetanus toxoid (TTE) and artificial epitope (PADRE), and 3 unique epitopes, aa1-20 of tubercle bacteria hot shock protein 65 (TBE), aa54-65 of rubella protein E2-4 (ME) and aa35-48 of trachoma hot shock protein 60 (CE) were chosen. Eukaryotic expression vectors were constructed by inserting single or multiple exogenous epitopes in HBV S gene just after the initial code of translation. BALB/c mice were inoculated with 100 micro g of recombinant DNA per mouse, and given boost inoculation for 3 times with 3-week interval. Mouse blood were collected one month just after the third boost inoculation. Anti-HBs was detected using Abbott test kits.
HBV S eukaryotic gene expression vectors, pHB and 6 exogenous HTL epitope HBV S gene vectors, pHB-TBE, pHB-PADRE, pHB-TTE, pHB-MTE2, pHB-MTE3 and pHB-MTE5 were constructed successfully with anti-HBs level (IU/L) of 10 +/- 5, 5 +/- 5, 49 +/- 7, 29 +/- 6, 16 +/- 8, 23 +/- 7 and 28 +/- 8 respectively. Among 3 single epitopes, TTE and PADRE had obviously effect on promoting the anti-HBs response of HBV S gene, while TBE had no promoting effect. All of the 3 multiple epitopes were shown the effect of immune promoting.
Some exogenous HTL epitopes had obviously effect on promoting the anti-HBs response of HBV S gene. Multiple epitopes also had humoral immunity promoting effect, but there was no synergic effect among their own HTL epitopes. PADRE might be an important candidate for new efficient HB vaccine. The multiple epitope cluster consisted form 5 exogenous epitopes might be an important candidate for the reinoculating HB vaccine or therapy HB vaccine.
研究辅助性T淋巴细胞（HTL）外源性表位对乙肝病毒S基因DNA免疫体液免疫的影响。
选择两个通用的HTL表位，破伤风类毒素的氨基酸残基（aa）830 - 843（TTE）和人工表位（PADRE），以及3个独特表位，结核杆菌热休克蛋白65的aa1 - 20（TBE）、风疹蛋白E2 - 4的aa54 - 65（ME）和沙眼热休克蛋白60的aa35 - 48（CE）。通过将单个或多个外源性表位插入乙肝病毒S基因翻译起始密码子之后构建真核表达载体。每只BALB/c小鼠接种100μg重组DNA，并每隔3周加强接种3次。在第三次加强接种后1个月采集小鼠血液。使用雅培检测试剂盒检测抗-HBs。
成功构建了乙肝病毒S真核基因表达载体pHB以及6个外源性HTL表位乙肝病毒S基因载体，pHB - TBE、pHB - PADRE、pHB - TTE、pHB - MTE2、pHB - MTE3和pHB - MTE5，其抗-HBs水平（IU/L）分别为10±5、5±5、49±7、29±6、16±8、23±7和28±8。在3个单表位中，TTE和PADRE对促进乙肝病毒S基因的抗-HBs反应有明显作用，而TBE无促进作用。所有3个多表位均显示出免疫促进作用。
一些外源性HTL表位对促进乙肝病毒S基因的抗-HBs反应有明显作用。多表位也有体液免疫促进作用，但它们自身的HTL表位之间没有协同作用。PADRE可能是新型高效乙肝疫苗的重要候选物。由5个外源性表位组成的多表位簇可能是再次接种乙肝疫苗或治疗性乙肝疫苗的重要候选物。
