Nehashi K, Yoshida J, Wakabayashi T, Nagata M, Utsumi J, Naruse N, Sugita K
Department of Neurosurgery, Nagoya University School of Medicine, Japan.
Neurol Med Chir (Tokyo). 1995 Oct;35(10):719-22. doi: 10.2176/nmc.35.719.
Superinduction of human interferon-beta (HuIFN-beta) from human glioma cells has greater cytotoxicity than purified HuIFN-beta derived from fibroblasts. However, superinduction requires several reagents like polyI:polyC, cycloheximide, and actinomycin D, which may contaminate the conditioned medium and obscure the effect of superinduced HuIFN-beta. The present study used minimum doses of polyI:polyC and cycloheximide without actinomycin D to superinduce HuIFN-beta. The superinduced HuIFN-beta was purified by passing the medium through molecular sieve column chromatography. Fractionation of the eluate provided semipurified superinduced HuIFN-beta which demonstrated a growth inhibitory effect against both the U251-MG autologous human glioma cell line and the SK-MG-1 homologous glioma cell line. This effect was neutralized by addition of anti-HuIFN-beta monoclonal antibody (YSB-1). In a separate experiment, combinations of superinduction reagents were found not to have growth inhibitory effects because all inhibition in superinduced medium was completely neutralized by YSB-1. Superinduced HuIFN-beta has a pure growth inhibitory effect on both autologous and homologous glioma cells, so may affect autocrine secretion of cytokines.
人胶质瘤细胞超诱导产生的人β干扰素（HuIFN-β）比成纤维细胞来源的纯化HuIFN-β具有更强的细胞毒性。然而，超诱导需要几种试剂，如聚肌苷酸：聚胞苷酸、放线菌酮和放线菌素D，这些试剂可能会污染条件培养基并掩盖超诱导HuIFN-β的作用。本研究使用最小剂量的聚肌苷酸：聚胞苷酸和放线菌酮，不使用放线菌素D来超诱导HuIFN-β。通过将培养基通过分子筛柱色谱法来纯化超诱导的HuIFN-β。洗脱液的分级分离提供了半纯化的超诱导HuIFN-β，其对U251-MG人自体胶质瘤细胞系和SK-MG-1同源胶质瘤细胞系均表现出生长抑制作用。通过添加抗HuIFN-β单克隆抗体（YSB-1）可中和这种作用。在另一项实验中，发现超诱导试剂的组合没有生长抑制作用，因为超诱导培养基中的所有抑制作用都被YSB-1完全中和。超诱导的HuIFN-β对自体和同源胶质瘤细胞均具有纯粹的生长抑制作用，因此可能影响细胞因子的自分泌。
