Tα1对食管癌细胞RNA转染的脐血树突状细胞诱导T细胞杀瘤活性的影响

Effect of Tα1 on ESCC RNA-transfected human cord blood dendritic cells induced antitumor activity of cytotoxic T lymphocytes

目的:探讨食管癌细胞RNA转染脐血树突状细胞(dendriti ccells,DCs)对T细胞增殖和CTL特异性抗肿瘤作用的影响,以及Tα1对脐血DCs疫苗的免疫佐剂作用.方法:分离脐血单个核细胞,经rhSCF、rhGM-CSF和rhIL-4诱导和T.Tn细胞RNA转染形成成熟DCs,加入Tα1制备肿瘤疫苗.流式细胞术(FCM)检测各组DCs表型变化;MTT法检测各组脐血DCs诱导T细胞增殖和CTL细胞毒活性.结果:T.Tn细胞RNA转染后组较转染前组脐血DCs高表达MHC-Ⅰ、MHC-Ⅱ、CD54、CD80和CD86分子(MHC-Ⅰ:70.36±6.09vs8.17±1.93;MHC-Ⅱ:72.03±5.32vs7.64±5.33;CD54:69.36±7.33vs2.05±2.03;CD80:67.21±6.77vs2.33±1.65;CD86:68.85±7.41vs6.73±1.97,P<0.01);与转染前组相比,可显著促进T细胞增殖(10∶1:4.77±0.79vs1.65±0.71;50∶1:3.85±0.57vs1.56±0.13;100∶1:2.89±0.59vs1.19±0.21,P<0.05),并有效诱导CTL的特异性杀瘤活性(10∶1:27.36±8.93vs10.35±2.93;20∶1:44.55±2.36vs11.77±1.03;50∶1:51.08±4.92vs12.75±1.49,P<0.05).而Tα1能显著促进RNA致敏脐血DCs的各种表面分子表达(MHC-Ⅰ:87.88±9.13vs70.36±6.09;MHC-Ⅱ:93.16±3.34vs72.03±5.32;CD54:91.75±3.84vs69.36±7.33;CD80:87.27±8.68vs67.21±6.77;CD86:89.09±6.86vs68.857.41,P<0.05);和刺激T细胞增殖能力(10∶1:8.31±1.78vs4.77±0.79;50∶1:5.97±0.14vs3.85±0.57;100∶1:4.03±0.13vs2.89±0.59,P<0.05);及诱导CTL的能力(10∶1:47.66±4.12vs27.36±8.93;20∶1:56.72±7.24vs44.55±2.36;50∶1:76.48±3.47vs51.08±4.92,P<0.05).结论:Tα1联合食管癌细胞RNA转染脐血DCs可制备高效、特异的肿瘤疫苗,有望成为食管癌生物治疗的新途径.

AIM： To investigate effect of human cord blood dendritic cells （DCs） transfected with RNA of esophageal carcinoma cells on the proliferation of T lymphocyte and on the antitumor activity of cytotoxic T lymphocytes （CTL）, and immunoadjuvant effect of T^I on human cord blood dendritic cells. METHODS： The RNA of T.Tn cells were prepared by TRIzol reagent. The cord blood monocytes were cultured to produce DCs with rhSCF,rhGM-CSF and rhlL-4. The DCs were collected and transfected with tumor cell total RNA. Tα1 was added in culture system to enhance the DCs vaccine. MHC-Ⅰ, MHC-Ⅱ and co-stimulatory molecules, CD54, CD80 and CD86 on the surface of DCs were analyzed by FCM. The mixed lymphocyte reaction （MLR） and cytotoxicity of CTLs to T.Tn cells was assayed using MTT colorimetry. RESUITS： Compared with pre-transfection group, expressions of MHC-Ⅰ, MHC-Ⅱ, CD54, CD80 and CD86 （MHC-Ⅰ： 70.36 ± 6.09 vs 8.17 ± 1.93; MHC-Ⅱ： 72.03 ± 5.32 vs 7.64 ± 5.33; CD54：69.36 ± 7.33 vs 2.05 ± 2.03; CD80：67.21 ± 6.77 vs 2.33 ± 1.65; CD86：68.85 ± 7.41 vs 6.73 ± 1.97, P 〈 0.01） were significantly higher in DCs transfected with RNA of esophageal carcinoma cells; T cell proliferation was markedly enhanced （10 ： 1：4.77 ± 0.79 vs 1.65 ± 0.71; 50 ： 1：3.85 ± 0.57 vs 1.56 ± 0.13; 100 ： 1：2.89 ± 0.59 vs 1.19 ± 0.21, P 〈 0.05）; and tumor cytolytic activities of cytotoxic T lymphocyte were effectively induced （10 ： 1：27.36 ± 8.93 vs 10.35 ± 2.93; 20 ： 1：44.55 ± 2.36 vs 11.77 ± 1.03; 50 ： 1：51.08 ± 4.92 vs 12.75 ± 1.49, P 〈 0.05）. Tα1 enhanced significantly the surface molecule expression （MHC-Ⅰ： 87.88 ± 9.13 vs 70.36 ± 6.09; MHC-Ⅱ： 93.16 ± 3.34 vs 72.03 ± 5.32; CD54：91.75 ± 3.84 vs 69.36 ± 7.33; CD80：87.27 ± 8.68 vs 6Z21 ± 6.77; CD86：89.09 ± 6.86 vs 68.85 ± 7.41, P 〈 0.05）, stimulation of proliferation （10 ： 1： 8.31 ± 1.78 vs 4.77 ± 0.79; 50 ： 1：5.97 ± 0.14 vs 3.85 ± 0.57; 100 ： 1：4.03 ± 0.13 vs 2.89 ± 0.59, P 〈 0.05） and activity of cytotoxic T lymphocytes （10 ： 1：47.66 ± 4.12 vs 27.36 ± 8.93; 20 ： 1：56.72 ± 7.24 vs 44.55 ± 2.36; 50 ： 1：76.48 ± 3.47 vs 51.08 ± 4.92, P 〈 0.05）. CONCLUSION： DCs from human cord blood monocytes exhibit high expression of MHC and co-stimulatory molecules and enhances T lymphocyte capability after transfection with RNA of esophageal carcinoma. The vaccine of cord blood DCs with adjuvant of Tα1 may provide an effective and specific way for immunotherapy of esophageal carcinoma.