GPC3特异性HLA-A11限制的T淋巴细胞表位肽免疫活性检测

Immune activity evaluation of GPC3 peptides recognized by HLA-A11 restricted T lymphocytes in hepatocellular carcinoma patients

目的设计合成人类白细胞抗原(HLA)-A11限制性T细胞识别的磷脂酰肌醇蛋白聚糖3(GPC3)多肽,并检测其免疫原性和反应性。方法利用BIMAS和SYFPEITHI评分系统评价GPC3多肽与HLA-A11分子的结合能力,预测出相应的抗原谱,合成HLA-A1101限制性GPC3多肽库;流式细胞仪检测HCC患者HLA-A11分子的表达;多肽刺激外周血淋巴细胞,酶联免疫斑点试验(ELISPOT)检测T细胞对GPC3多肽的反应性。结果流式细胞仪检测32例HCC患者,7例HLA-A11分子表达阳性(21.9%);ELISPOT检测发现17例(53.1%)HCC患者的T细胞应答阳性,7例(21.9%)HLA-A11表型检测阳性的HCC患者,T细胞应答结果成强阳性。HLA-A11限制性GPC3多肽应答率符合人群中相应基因位点的自然表达率。结论 GPC3多肽设计合理,具有良好的免疫原性和反应性。

Objective To design and synthesize GPC3 peptides recognized by HLA-A11- restricted T lymphocytes, and to detect their immunogenicity and reactivity. Methods The GPC3 peptides and HLA-A11 molecules were evaluated, and the corresponding antigen spectrum was predicted by BIMAS and SYFPEITHI scoring system.HLA-A1101 which restricted GPC3 peptide library was synthesized; the expression of HLA-A11 molecules in hepatocelllular carcinoma（HCC） patients was tested by flow cytometry; peripheral blood lymphocytes were stimulated with peptides,and reactive T lymphocytes were detected with ELISPOT for secretion of IFN-γ. Results Thirty-two HCC patients were detected by flow cytometry for HLA-A11 molecular expression with 7 cases（21.9%） positive. By ELISPOT, 17 cases were detected for T lymphocytes respose with positive. Meanwhile, the T lymphocytes responses in the 7 HCC patients were detected as strongly positive, showing that the HLA-A11 restricted GPC3 peptide response rate was consisted with the natural expression rate of the corresponding gene locus. Conclusion The GPC3 polypeptide is reasonable designed with good immunogenicity and reactivity.