鼻咽癌可溶性抗原负载DC诱导CTL杀伤鼻咽癌细胞的体外研究

目的：用负载鼻咽癌细胞(CNE)可溶性抗原的树突状细胞(dendritic cells，DC)诱导自身淋巴细胞，使之活化为鼻咽癌(nasopharyngeal carcinoma，NPC)特异性细胞毒性T细胞(cvtotoxic T lymphocyte，CTL)，观察其对鼻咽癌细胞的体外特异性杀伤作用。方法：用GM-CSF、IL-4和TNF-α体外诱导健康成人外周血单核细胞，使其分化为高纯度DC。用鼻咽癌细胞(CNE)可溶性抗原负载成熟DC，流式细胞仪检测负载前后DC的表型特征，MLR检测负载前后DC对同种异体T淋巴细胞增殖的能力。在白细胞介素-2(IL-2)的作用下，用负载鼻咽癌细胞可溶性抗原的DC诱导自身淋巴细胞成NPC特异性CTL，通过杀伤活性实验观察它对鼻咽癌细胞(CNE)的特异性杀伤效应。结果：人外周血单核细胞体外在GM-CSF、IL-4、TNF-α诱导下获得具典型表型特征的成熟DC。负载CNE可溶性抗原对成熟DC表面共刺激分子及特异性表面标志无影响，仍有较强的刺激同种异体淋巴细胞增殖能力，所诱导的CTL对鼻咽癌细胞(CNE)有明显的特异性杀伤作用。结论：负载鼻咽癌细胞(CNE)可溶性抗原的DC所诱导的CTL在体外对鼻咽癌细胞有特异性杀伤作用，对NPC的临床治疗有潜在的应用价值。

Cytotoxic T-lymphocytes induced by dendritic cells loaded with nasopharyngeal carcinoma epithelioid cell lines (CNE) kills CNE cells in vitro

Objective:To study the killing effect of cytotoxic T-lymphocytes (CTLs) after activated by dendritic cells (DCs) loaded with soluble antigens of CNE upon CNE in vitro.Methods: The monocytes isolated from healthy human peripheral blood were cultured with granulocyte-monocyte colony-stimulating factor (GM-CSF) and interleukin-4(IL-4) for 7 days. Before and after loaded with soluble antigens of CNE,the phenotype of mature DC was detected and the function of DC to stimulate allogeneic cells proliferation was measured. Host lymphocytes were stimulated with DCs loaded with soluble antigen of CNE in the culture medium containing interleukin-2 (IL-2) for 7 days. The special lysis activity of CTLs against nasopharyngeal carcinoma CNE cells was analysed. Results: High purity mature DCs displaying typically characteristics of phenotype were obtained from monocytes. After being loaded with soluble antigens of CNE cells, mature DCs still had strong potential to stimulate the proliferation of allogenetic T cells. No significant changes in the surface markers and the cytomorphology of mature DCs were detected during the loading. Activated lymphocytes strongly showed special cytotoxic ability against nasopharyngeal carcinoma CNE cells.Conclusion: The CTLs induced by DC loaded with soluble antigens of CNE cells have a killing effect on nasopharyngeal carcinoma cells in vitro,which has the therapeutic potentials for nasopharyngeal carcinoma.