GPC3特异性HLA-A11限制的T淋巴细胞表位肽免疫活性检测

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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 a glypican 3 (GPC3) polypeptide recognizing human HLA-A11 restricted T cells and test its immunogenicity and reactivity. Methods BIMAS and SYFPEITHI scoring system was used to evaluate the binding ability of GPC3 peptides to HLA-A11 molecules, and the corresponding antigen spectrum was predicted to synthesize HLA-A1101 restricted GPC3 polypeptide library. The expression of HLA-A11 in HCC patients was detected by flow cytometry. Polypeptides stimulate peripheral blood lymphocytes, ELISPOT test T cells to GPC3 polypeptide reactivity. Results Twenty-two patients with HCC were detected by flow cytometry. The positive expression of HLA-A11 was detected in 7 of 7 (21.9%) patients by ELISPOT. T-cell responses were positive in 17 patients (53.1% HCC patients with positive phenotype test result strongly positive for T cell response. The HLA-A11 restricted GPC3 polypeptide response rate is in line with the natural rate of the corresponding gene locus in the population. Conclusion The GPC3 peptide has reasonable design, good immunogenicity and reactivity.
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