摘要
CTL是红前期恶性疟免疫防护的关键环节。MHC分子多态性和严格的种属特异性是限制CTL恶性疟重组疫苗研制的重要因素。本文选用中国人群常见的HLAI类分子A-2．1和B51限制的恶性疟CTL抗原表位TR26和SH6，进行表位DNA序列合成并将其克隆于真核表达载体，构建单表位DNA重组疫苗。同时应用同尾酶克隆技术，将上述表位DNA序列串联构建成双表位DNA重组疫苗pcDNA3．1TS。将以上克隆在含相应HLA抗原分子的细胞株中进行表达。经细胞表面HLAⅠ类分子表达水平检测证实，两个单表位DNA疫苗编码的短肽在细胞内的表达，明显促进相应HLAⅠ类分子在细胞表面的表达，用流式细胞仪测定平均荧光强度可量化表达水平（P＜0．05）。串联双表位DNA疫苗编码的肽并不受其位置毗邻的影响，分别促进HLA-A0201和HLA-B51的表达（P＜0．05），提示各自在相应细胞株内被加工违呈，此项研究提示：该串联表位疫苗可能为两种MHC遗传背景的人群提供免疫防护。
CD8+ cytotoxic T lymphocytes (CTL) play a critical role in eradicating malaria at the pre-erythrocyte stageof the parasite infection. Some obstacles to the development of a CTL vaccine include the polymorphism of MHC Imolecules and their restrictions to the presentation of antigens. In this study, two Plasmodium falciparum CTLepitopes which was restricted to Chinese population common HLA class I molecules: HLA-B51 and HLA-A2.1 respectively were chosen and cloned separately into an eukaryotic expressing vector to form pcDNA3. 1 SH6, pcDNA3. 1 TR26,and pcDNA3.1TS, which had the two aforementioned epitopes fused in tandem. All of these clones were transfectedand expressed in respective cell lines containing appropriate HLA molecules. An obviously increased expressions ofHLA class I molecules were detected in the transfected cell lines, evaluated as mean channel number of fluorescenceby flow cytometry (P<0.05). It was demonstrated that the two discrete Plasmodium falciparum CTL epitope as inrecombinant vaccine pcDNA3. 1 SH6 and pcDNA3. 1 TR26 were effectively processed and presented, and the closeproximity of the two epitope in one chain as in recombinant vaccine pcDNA3. 1 TS did not inteffere with the processingand presenting of each epitope gene in corresponding cell line. This work suggested the multi-epitope vaccine mightprovide protection for two populations which containing different MHC class I background.
出处
《基础医学与临床》
CSCD
2000年第3期32-37,共6页
Basic and Clinical Medicine
基金
国家自然科学基金!39770670
美国中华医学基金会(CMB)资助!98-674
关键词
恶性疟
多表位疫苗
细胞毒T淋巴细胞
HLA-B51
plasmodium falciparum multi-epitope vaccine
cytotoxic T lymphocyte
human leukocyte antigen class Ⅰ molecule
