摘要
目的研究人巨细胞病毒(hCMV)结构蛋白pp65衍生抗原肽(pp65495503,NLV)特异性CD8+T细胞的功能特性。方法将HLAA2NLV四聚体染色与细胞内细胞因子或穿孔素染色相结合,以流式细胞仪直接分析NLV特异性CD8+T细胞内穿孔素的表达水平,或者在NLV抗原肽刺激6h后分析γ干扰素(IFNγ)的表达水平。结果NLV抗原肽能诱导hCMV特异性CD8+T细胞分泌IFNγ,不同供者特异性CD8+T细胞产生IFNγ的比率存在较大差异(55.69±17.64)%,当NLV抗原肽质量浓度为10μgmL时IFNγ阳性细胞百分率最高;未经刺激的特异性CD8+T细胞内表达较高水平的穿孔素(53.90±16.41)%。结论识别单一表位的hCMV特异性CD8+T细胞合成IFNγ和穿孔素的能力存在多样性。
Objective To investigate the functional characteristics of CD8^＋ T cells specific to human cytomegalovims （hCMV） structural protein pp65-derived peptide antigen （pp65495-503, NLV）. Methods Perforin expression without stimulation was analyzed with flow cytometry by using HLA-A2/NLV tetramer staining together with intracellular staining, while γ-interferon （IFN-γ） expression was evaluated by using tetramer staining in combination with intracellular cytokine staining after stimulation with NLV peptide for 6 h. Results IFN-γ was expressed in hCMV-specific CD8^＋ T cells after induction of the NLV peptide, and the expression levels of IFN-γ in different donors were highly variable [ （55.69±17.64）% ]. The optimal concentration of the NLV peptide was 10μg/mL. Moreover, perforin was expressed at high level [ （53.90 ±16.41 ） % ] within unstimulated antigen-specific CD8^＋ T cells. Conclusion The hCMV-specific CD8^＋ T cells recognizing single epitope are heterogeneous in synthesizing IFN-γ and perforin.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2006年第4期440-443,共4页
Immunological Journal
基金
国家自然科学基金重点项目(30230350)
国家自然科学基金面上项目(30371651
30572199)资助
