Ishikawa Eiichi, Tsuboi Koji, Saijo Kaoru, Harada Hideki, Takano Shingo, Nose Tadao, Ohno Tadao
Department of Neurosurgery, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba Science City, Ibaraki 305-8575, Japan.
Anticancer Res. 2004 May-Jun;24(3b):1861-71.
Natural killer (NK) cells are highly efficient in the cellular immune response against malignant tumors without restriction of major histocompatibility complex. However clinical studies using autologous NK cells have been reported in only a very limited number of cases, due to the fact that selective NK expansion is difficult to achieve in this patient population. Here, we report the results of adoptive immunotherapy in patients with recurrent malignant gliomas using autologous NK cells that were expanded ex vivo by a novel method.
Peripheral blood mononuclear cells (PBMCs) were prepared from patients with malignant gliomas, and were co-cultured with an irradiated human feeder cell line (HFWT) in RHAM-alpha medium supplemented with 5% autologous plasma and interleukin-2. The resulting NK cell-rich effector cells were injected into 9 patients (16 courses) with recurrent malignant glioma (6 cases of WHO grade-3 glioma and 3 cases of grade-4 glioma).
The mean frequency of NK cells among lymphocytes was 82.2 +/- 10.5%. A combination of focal and intravenous injections was peformed in 10 courses. Intravenous injection alone was performed in 6 courses. Further, intravenous injection of low-dose interferon beta (6x10(6) IU/week) was performed as an adjuvant therapy in all courses to achieve maximum benefit for enrolled patients. Clinical evaluation demonstrated 3 PR, 2 MR, 4 NC and 7 PD in a total of 16 courses of treatment. Severe neurological toxicity was not observed in any of the patients.
It was demonstrated that NK cell-rich effector cells were expanded ex vivo from PBMCs in all nine cases of recurrent malignant glioma and that NK cell therapy was safe and partially effective in patients with recurrent malignant gliomas.
自然杀伤（NK）细胞在针对恶性肿瘤的细胞免疫反应中效率很高，不受主要组织相容性复合体的限制。然而，由于在这类患者群体中难以实现选择性NK细胞扩增，使用自体NK细胞的临床研究仅在非常有限的病例中得到报道。在此，我们报告了使用一种新方法在体外扩增的自体NK细胞对复发性恶性胶质瘤患者进行过继性免疫治疗的结果。
从恶性胶质瘤患者中制备外周血单个核细胞（PBMC），并在补充有5%自体血浆和白细胞介素-2的RHAM-α培养基中与经辐照的人饲养细胞系（HFWT）共培养。将产生的富含NK细胞的效应细胞注入9例（16疗程）复发性恶性胶质瘤患者（6例世界卫生组织3级胶质瘤和3例4级胶质瘤）。
淋巴细胞中NK细胞的平均频率为82.2±10.5%。10个疗程采用局部注射和静脉注射相结合的方式。仅静脉注射6个疗程。此外，在所有疗程中均进行静脉注射低剂量干扰素β（6×10⁶IU/周）作为辅助治疗，以使入组患者获得最大益处。临床评估显示，在总共16个疗程的治疗中，有3例部分缓解（PR）、2例疾病稳定（MR）、4例疾病无进展（NC）和7例疾病进展（PD）。所有患者均未观察到严重的神经毒性。
结果表明，在所有9例复发性恶性胶质瘤患者中，均可从PBMC体外扩增出富含NK细胞的效应细胞，且NK细胞疗法对复发性恶性胶质瘤患者安全且部分有效。
