MAGE-A3抗原肽负载树突状细胞诱导乳腺癌患者免疫应答的研究

目的研究MAGE-A3抗原肽负载对乳腺癌患者树突状细胞(DC)的功能的影响,探讨其是否可以在体外诱导特异性细胞毒性T淋巴细胞(CTL)应答.方法体外培养HLA-A2乳腺癌患者外周血来源的DC,并经孵育携带MAGE-A3抗原肽,用以刺激CTL,用3H-TdR渗入法检测抗原肽负载前后DC刺激同种淋巴细胞增殖能力(MLR),并用乳酸脱氢酶(LDH)释放法检测CTL对靶细胞的杀伤效应.结果DC:T为1:10时,单纯DC组刺激能力显著高于抗原肽负载组DC(DC-P)(P＜0.05),但当DC:T低于1:20时后DC-P组刺激能力显著强于单纯DC组(P＜0.05).DC-P组和单纯DC组所激发的CTL对细胞株MCF-7,Sk-Br-3,MDA-MB-435s的杀伤活性有显著性差异,而对细胞株Raji无显著性差异,DC-P组对细胞株MCF-7,Sk-Br-3,MDA-MB-435s的杀伤活性明显高于对细胞株Raji的杀伤活性.结论负载MAGE-A3抗原肽的DC在体外可以诱导乳腺癌患者特异性的CTL免疫应答,为临床以DC为基础的过继免疫治疗的应用提供理论基础.

Dendritic cells loaded with peptide derived from MAGE-A3 induced cytotoxic T-lymphocyte response in breast cancer patients in vitro

Objectve: To investigate whether the dendritic cells(DCs) loaded with the peptide derived from MAGE-A3 could induce specific cytotoxic T lymphocytes(CTLs) response in breast cancer patients in vitro. Methods: DCs were induced from peripheral blood mononuclear cells of 3 breast cancer patients with HLA-A2 phenotype. The DCs were pulsed by MAGE-A3 peptide(DC-P) were used to stimulated autologous T lymphocytes. Then the effect of the DCs stimulating the proliferation of autologous T lymphocytes was tested by 3H-TdR incorporation assay. The cytotoxicity of induced CTLs to various kinds of target cells was detected by the method of lactate dehydrogenase(LDH)releasing assay. Results: When effective cells: target cells(E:T)was 1:10,the stimulating effect of pure DCs is stronger than that of DC-P. But, when E:T was lower than 1:20, the stimulating effect of DC-P is stronger than that of DCs. There was significant difference between the cytotoxicity of DC-P and DCs to MCF-7,Sk-Br-3,MDA-MB-435s, but there was no significant difference between the cytotoxicity of DC-P and DCs to Raji. The cytotoxicity of DC-P to MCF-7,Sk-Br-3,MDA-MB-435s is significant stronger than that to Raji. Conlusion: The DC-P can elicit MAGE-A3 specific CTL responses in breast cancer patients in vitro. The results provide an experimental basis for clinical application of the adoptive immunotherapy based on DCs.