Sawayama Tomoyuki, Sakaguchi Kohsaku, Senoh Tomonori, Ohta Takeyuki, Nishimura Mamoru, Takaki Akinobu, Tsuji Takao, Shiratori Yasushi
Department of Medicine and Medical Science, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
Acta Med Okayama. 2003 Dec;57(6):285-92. doi: 10.18926/AMO/32813.
In patients with hepatocellular carcinoma (HCC), natural killer (NK) cell activity decreases significantly, and the reduced activity may be associated with the progression of HCC. In this study we evaluated the effects of pulsing with interleukin (IL)-2 and/or IL-12 on the activation of freshly isolated peripheral blood lymphocytes (PBL) derived from patients with HCC. PBL obtained from 9 HCC patients, 4 liver cirrhosis patients, and 9 normal subjects were cultured in the presence of IL-2 and/or IL-12. After 24 h of incubation, the levels of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha presented in the supernatants were determined by enzyme-linked immunosorbent assay (ELISA). The IFN-gamma and TNF-alpha production of PBL pulsed by a combination of IL-2 and IL-12 was significantly higher than those of PBL stimulated by either IL-2 or IL-12 alone. The mRNA encoding perforin, granzyme B, as well as IFN-gamma and TNF-alpha, were markedly enhanced in PBL stimulated with a combination of IL-12 and IL-2. The pulsing procedure of IL-12 in combination with IL-2 resulted in the increase of IFN-gamma and TNF-alpha, and the expression of perforin and granzyme B mRNA in PBL obtained from HCC patients, as well as in those obtained from normal subjects. These results indicate that adoptive immunotherapy based on PBL pulsed with a combination of IL-2 and IL-12 may be a promising adjunctive strategy for HCC treatment.
在肝细胞癌（HCC）患者中，自然杀伤（NK）细胞活性显著降低，而这种活性降低可能与HCC的进展有关。在本研究中，我们评估了用白细胞介素（IL）-2和/或IL-12刺激对新鲜分离的源自HCC患者的外周血淋巴细胞（PBL）激活的影响。从9例HCC患者以及4例肝硬化患者和9名正常受试者获取的PBL在IL-2和/或IL-12存在的情况下进行培养。孵育24小时后，通过酶联免疫吸附测定（ELISA）测定上清液中干扰素（IFN）-γ和肿瘤坏死因子（TNF）-α的水平。由IL-2和IL-12联合刺激的PBL产生的IFN-γ和TNF-α显著高于单独用IL-2或IL-12刺激的PBL。在用IL-12和IL-2联合刺激的PBL中，编码穿孔素、颗粒酶B以及IFN-γ和TNF-α的mRNA显著增强。IL-12与IL-2联合刺激的过程导致从HCC患者以及正常受试者获取的PBL中IFN-γ和TNF-α增加，以及穿孔素和颗粒酶B mRNA的表达增加。这些结果表明，基于用IL-2和IL-12联合刺激的PBL的过继免疫疗法可能是一种有前景的HCC辅助治疗策略。
