Clinical application of NKT cell immune system for lung cancer

我们在肺癌患者中进行了α半乳糖神经酰胺冲击的树突状细胞。对晚期非小细胞肺癌或复发肺癌患者静脉注射αGalCer冲击的DC(1x109/mm2)，以测试其安全性、可行性和临床疗效。所有完成病例均行免疫监测。符合条件的患者行外周血白细胞分离，采集外周血单个核细胞(PBMC)，密度梯度离心法进一步分离。PBMC与人粒细胞巨噬细胞集落刺激因子(GM-CSF)共同培养1~2周，并在给药前一天加入特异性配体α-半乳糖神经酰胺冲击。用流式细胞仪检测患者外周血中NKT细胞的频率，ELISPOT法检测γ-α再刺激后PBMC产生的干扰素。FN-γ产生明显增加的患者，其生存期和肿瘤进展时间相对较长，而α-γ产生反应差的患者预后很差。这是第一个显示干扰素-γ在NKT细胞免疫治疗中的关键作用的报告。根据这些结果，将对干扰素-γ的产生和抗肿瘤免疫反应的作用和机制进行讨论。

We performed αGalactosylceramide-pulsed Dendritic cells in patients with lung cancer. Patients with advanced non-small cell lung cancer or recurrent lung cancer received intravenous injections of αGalCer-pulsed DCs (1x10^9/m^2) to test the safety, feasibility, and clinical response. Immunomonitoring was also performed in all completed cases. Eligible patients underwent peripheral blood leukapheresis, and peripheral blood mononuclear cells (PBMCs) were collected and further separated by density gradient centrifugation. PBMCs were cultured with human granulocyte-macrophage colony-stimulate factor (GM-CSF) for 1-2 weeks and pulsed with specific ligand, α-Galactosylceramide on the day before administration.Seventeen patients were enrolled in this study. The frequency of peripheral blood NKT cells in all patients was measured by FACS analysis and IFN-γ production after restimulation with αGalCer in PBMCs was measured by ELISPOT assay. Patients with marked increase of FN-γ production following the administration of αGalCer-pulsed DC showed relatively long survival and time to tumor progression, while poor responder of IFN-γ production showed very poor prognosis. This is the first report to show the crucial role of IFN-γ at NKT cell immunotherapy. With these results, the role and mechanisms of IFN-γ production and anti-tumor immune response will be discussed.