Hironaka Katsuji, Yamaguchi Yoshiyuki, Okita Riki, Okawaki Makoto, Nagamine Ichiro
Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi 1-2-3, Minami-Ku, Hiroshima 734-8553, Japan.
Anticancer Res. 2006 Sep-Oct;26(5B):3701-7.
Toll-like receptors (TLRs) are important molecules that stimulate the innate immunity in order to eradicate microbial pathogens, after which the adaptive immunity emerges. The involvement of TLRs in the action mechanism of OK-432, a bacterial preparation, was investigated in the locoregional treatment of malignant ascites from gastric cancer. The expression of TLRs in ascites cells was analyzed using reverse-transcription polymerase chain reaction specific for TLRs and by flow cytometry using anti-TLR2, -TLR4, -CD4, -CD8, and -CD11c antibodies. These measurements were compared with the locoregional response of OK-432 immunotherapy for malignant ascites, as well as TNF-alpha producing potential, which was measured by ELISA, of ascites cells stimulated in vitro with OK-432. It was observed that OK-432 immunotherapy for malignant ascites showed 8 positive (67%) and 4 negative responses with the tolerable adverse effects of fever elevation and abdominal pain. The TNF-alpha production of ascites cells by in vitro OK-432 stimulation was significantly higher in responder patients than in non-responders. The clinical responses were correlated with the expression of the TLR4 gene of ascites cells. The TNF-alpha-producing potential of ascites cells by in vitro OK-432 stimulation was dependent on the existence of a CD11c + TLR-4+ cell population in ascites cells. OK-432 was highly stimulatory for TNF-alpha production of ascites cells compared with other biological response modifiers of PSK and LEM. These results suggest that TLR-4 expression on ascites cells of a macrophage lineage is essential for ascites cells to produce TNF-alpha in relation to OK-432 stimulation and for subsequent positive clinical responses in locoregional immunotherapy using OK-432 for malignant ascites from gastric cancer.
Toll样受体（TLRs）是刺激先天免疫以根除微生物病原体的重要分子，之后适应性免疫才会出现。我们研究了TLRs在细菌制剂OK-432作用机制中的参与情况，该研究针对胃癌恶性腹水的局部区域治疗。使用针对TLRs的逆转录聚合酶链反应以及通过使用抗TLR2、-TLR4、-CD4、-CD8和-CD11c抗体的流式细胞术分析腹水细胞中TLRs的表达。将这些测量结果与OK-432免疫疗法对恶性腹水的局部区域反应以及用OK-432体外刺激的腹水细胞通过酶联免疫吸附测定法测量的产生肿瘤坏死因子-α（TNF-α）的潜力进行比较。观察到OK-432对恶性腹水的免疫疗法显示8例阳性（67%）和4例阴性反应，伴有发热升高和腹痛等可耐受的不良反应。体外OK-432刺激下，反应者患者腹水细胞产生的TNF-α明显高于无反应者。临床反应与腹水细胞TLR4基因的表达相关。体外OK-432刺激下腹水细胞产生TNF-α的潜力取决于腹水细胞中CD11c + TLR-4+细胞群的存在。与PSK和LEM的其他生物反应调节剂相比，OK-432对腹水细胞产生TNF-α具有高度刺激作用。这些结果表明，巨噬细胞谱系腹水细胞上的TLR-4表达对于腹水细胞在OK-432刺激下产生TNF-α以及随后使用OK-432对胃癌恶性腹水进行局部区域免疫治疗的阳性临床反应至关重要。
