Ren Wei-na, Chang Chun-kang, Fan Hua-hua, Guo Fang, Ren Ya-na, Yang Jie, Guo Juan, Li Xiao
Department of Hematology, The Sixth People's Hospital, Shanghai Jiaotong University, Shanghai, PR China.
J Immunoassay Immunochem. 2011;32(3):207-18. doi: 10.1080/15321819.2011.559295.
To improve its antitumor effect, we used human leukocyte antigen -A2 (HLA-A2)-positive human dendritic cell (DC)-derived DEXs (DC-derived exosomes) to support NY-ESO-1 antigen and polyI:C, with the aim of increasing the proliferation of specific cytotoxic T lymphocytes (CTL) in transgenic mice. Mature dendritic cells derived from peripheral blood mononuclear cells (PBMC) were isolated from the blood of healthy adults with positive HLA-2A. Using centrifuge and membrane ultrafiltration, EXO (exosomes) were extracted from the supernatant of DCs secretions. Transgenic C57 mice were immunized with human-derived tumor testis antigen NY-ESO-1/EXO, with or without polyI:C. Mice were sacrificed four weeks after immunization, and spleen cells were isolated and tested for function. The experiments included antigen-specific CTL proliferation, as tested by dimerization and antitumor effects for K562 cells as well as melanoma, tested at different ratios of effected cells:target cells (0:1, 10:1, 50:1, and 100:1). Dimerization experiments indicated that the effect of DEX/TSA (tumor specific antigens) + PolyI:C was 2.36 ± 1.10% and the control was 0.38 ± 0.31%, while the effect of DEX/TSA was 1.97 ± 0.63% and the control was 0.36 ± 0.07%. Antitumor effects by DEX/TSA: PolyI:C for the cell ratios of 0:1, 10:1, 50:1, and 100:1 were 11.14 ± 1.36%, 14.17 ± 0.62%, 15.71 ± 2.48%, and 24.31 ± 2.91%, respectively, for K562 cells. The antitumor effects for DEX/TSA for the cell ratios of 0:1, 10:1, 50:1, and 100:1 were 12.23 ± 2.25%, 13.10 ± 1.57%, 15.27 ± 2.93%, and 19.87 ± 2.72%, respectively, for K562 cells. With ratios of 10:1 and 100:1, the antitumor effects of DEX/TSA + PolyI:C were better than for the DEX/TSA group (P < 0.05). However, higher ratios of effecter cells to target cells increased, and there were no significant improvements in antitumor effect for control cells. Combining PolyI:C with DEX/TSA derived from healthy human blood positive for HLA-A2 is a promising strategy for developing new subcellular antitumor vaccination.
为提高其抗肿瘤效果，我们使用人白细胞抗原 -A2（HLA -A2）阳性的人树突状细胞（DC）来源的DEXs（DC衍生的外泌体）负载NY - ESO - 1抗原和聚肌胞苷酸，目的是增加转基因小鼠中特异性细胞毒性T淋巴细胞（CTL）的增殖。从HLA - 2A阳性的健康成年人血液中分离出源自外周血单核细胞（PBMC）的成熟树突状细胞。通过离心和膜超滤从DC分泌物的上清液中提取EXO（外泌体）。用源自人的肿瘤睾丸抗原NY - ESO - 1/EXO免疫转基因C57小鼠，同时添加或不添加聚肌胞苷酸。免疫四周后处死小鼠，分离脾细胞并检测其功能。实验包括通过二聚化检测抗原特异性CTL增殖，以及在不同效应细胞：靶细胞比例（0:1、10:1、50:1和100:1）下检测对K562细胞以及黑色素瘤的抗肿瘤作用。二聚化实验表明，DEX/TSA（肿瘤特异性抗原）+聚肌胞苷酸组的效应为2.36±1.10%，对照组为0.38±0.31%，而DEX/TSA组的效应为1.97±0.63%，对照组为0.36±0.07%。对于K562细胞，DEX/TSA:聚肌胞苷酸在细胞比例为0:1、10:1、50:1和100:1时的抗肿瘤作用分别为11.14±1.36%、14.17±0.62%、15.71±2.48%和24.31±2.91%。对于K562细胞，DEX/TSA在细胞比例为0:1、10:1、50:1和100:1时的抗肿瘤作用分别为12.23±2.25%、13.10±1.57%、15.27±2.93%和19.87±2.72%。在比例为10:1和100:1时，DEX/TSA +聚肌胞苷酸组的抗肿瘤作用优于DEX/TSA组（P<0.05）。然而，效应细胞与靶细胞的比例升高时，对照细胞的抗肿瘤作用没有显著改善。将聚肌胞苷酸与源自HLA - A2阳性健康人血液的DEX/TSA联合使用是开发新型亚细胞抗肿瘤疫苗的一种有前景的策略。
