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目的探索WT1肽体外抗肿瘤效应。方法合成含HLA-A*0201锚定位点的9个氨基酸的WT1-235肽。PCR-SSP法筛选HLA-A*0201健康供者并鉴定细胞株HLA-A基因型别,RT-PCR鉴定靶细胞株WT1表达与否。体外分离HLA-A*0201型别的外周血单个核细胞(PBMCs),培养树突状细胞(DC),负载WT1肽并刺激活化同源淋巴细胞,进行体外CTLs杀伤靶细胞的细胞毒试验。结果荷肽DC活化的CTLs对高表达WT1的HLA-A*0201型别细胞株SW48的杀伤效应高于不表达WT1的HLA-A*0201型别的人T细胞,也高于表达WT1但非HLA-A*0201型别的K562细胞(P<0.05)。结论 WT235-242肽具有体外抗肿瘤效应并受MHC限制。
Objective To explore the anti-tumor effect of WT1 peptide in vitro. Methods A 9 amino acid WT1-235 peptide containing the HLA-A * 0201 anchor site was synthesized. The HLA-A * 0201 healthy donors were screened by PCR-SSP and HLA-A genotypes were identified. RT-PCR was used to identify the expression of WT1 in the target cell lines. In vitro, HLA-A * 0201 type PBMCs were isolated from human peripheral blood mononuclear cells (PBMCs), cultured DCs, loaded with WT1 peptide and stimulated activated lymphocytes. CTLs were used to kill target cells in vitro. Results CTLs with DCs activated by peptide were more effective than HLA-A * 0201 cell line SW48 with high expression of WT1, but higher than WT T cells expressing HLA-A * 0201 without expressing WT1 HLA-A * 0201-type K562 cells (P <0.05). Conclusion WT235-242 peptide has anti-tumor effect in vitro and is limited by MHC.