中文摘要：

目的探讨纯化的隐球菌抗原肽GMFDGLSGV二聚体、四聚体、八聚体重组蛋白激发细胞毒性T淋巴细胞（Cytotoxic T lymphocyte,CTL）免疫应答的能力,为新型隐球菌疫苗的研制打下基础。方法通过细胞增殖和细胞毒性实验,研究纯化的GMFDGLSGV单体、二聚体、四聚体、八聚体重组蛋白刺激外周血单个核细胞（PBMC）产生的增殖反应和细胞毒活性,初步鉴定重组多聚体的免疫原性。结果10例HLA-A＊0201阳性个体PBMC对八聚体的平均SI为37.4±3.07;增殖反应明显高于四聚体（平均SI为20.1±1.37）、二聚体（平均SI为14.1±1.06）、单体（平均SI为11.4±0.68）以及阴性对照（平均SI为1±0.10）引起的细胞增殖反应（P〈0.01）。在抗原肽诱导的CTL细胞毒活性中,HLA-A＊0201阳性个体PBMC对八聚体、四聚体、二聚体、单体及阴性对照组均表现不同程度的杀伤活性,平均活性分别为48.1±4.28、23.9±2.20、16.3±1.27、13.1±1.10、1.0±0.1（P〈0.01）。结论本研究得到的重组优化八聚体具有较好的免疫原性,能有效刺激PBMC产生细胞增殖反应和细胞毒活性,为进一步开发有效的隐球菌疫苗打下了基础。

英文摘要：

Objective To investigate immunogenicity of muhimer peptides from mycobacterium tuberculosis （ Mth）, Methods Cellular proliferation and cytotoxicity experiments were performed to evaluate the immunogenicity of recombinant muhimer peptides from Mtb. Results Multimer peptides （ 10μM） could stimulate proliferation of peripheral blood monocytes （PBMCs） from all of 14 HLA-A ＊ 0201 positive （ A2 ^＋ ） PPD^＋ healthy candidates, strikingly stronger than those of single peptide＇ s and PPD＇ s groups （ average stimulation index as 37.4 ± 3.07 versus 20.1± 1.37 or 14.1±1.06 or 11.4±0.68 or 1± 0.10, respectively, P 〈 0.01 ）, CTLs induced by multimer peptides were used as effector cells in a CTL assay against T2 cells loaded with single peptide （ 10 μM） Multimer peptides could induce CTL cytotoxic activity in all of the 14 A2^＋ PPD ^＋ healthy candidates, significantly stronger than single peptide＇ s and PPD＇ s groups （ average activity as 48.1 ± 8.9% versus 23.9 ± 9.2% or16.3 ± 7.8% or 13.1 ± 8.4% or 1 ± 10%, respectively, P 〈 0.01 ）. Conclusion The recombinant octamerous peptide from Mth can stimulate greater proliferation and cytotoxicity of PBMCs, which is helpful for designing new vaccine.