Gene transfer of a hybrid interleukin-1 beta gene to B16 mouse melanoma recruits leucocyte subsets and reduces tumour growth in vivo
Gene transfer of a hybrid interleukin-1 beta gene to B16 mouse melanoma recruits leucocyte subsets and reduces tumour growth in vivo
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DOI：
10.1007/s002620050383
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发表时间：
1997-07-01
影响因子：
5.8
通讯作者：
Dohlsten, M
中科院分区：
文献类型：
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作者：
Bjorkdahl, O;Wingren, AG;Dohlsten, M
白细胞介素（IL）-1与大多数其他细胞因子的不同之处在于它缺乏信号序列。这导致未成熟预成型物的细胞内滞留。已显示IL-1的释放主要限于活化的单核细胞和巨噬细胞，并与生产细胞的凋亡有关。这些特征限制了IL-1在早期免疫应答中的研究。为了研究抗肿瘤免疫应答过程中局部IL-1 β释放的生物学效应，我们使用了用成熟人IL-1 β cDNA构建体转导的B16小鼠黑素瘤细胞。为了获得释放形式的人IL-1 β（ssIL-1 β），将来自相关IL-1受体拮抗剂的信号序列连接到编码成熟形式IL-1 β的cDNA上。当免疫原性差的B16黑色素瘤细胞系的细胞通过逆转录病毒感染用IL-1 β转导时，在细胞内检测到高水平的蛋白质，而用含有信号序列的IL-1 β转导的细胞分泌它们的大部分蛋白质。黑色素瘤细胞的体外生长不受IL-1 β或ssIL-1 β基因转移的影响。相比之下，与IL-1 β和模拟物转导的对照相比，同系C57 BL/6小鼠中的ssIL-1 β转导的B16细胞的体内皮下肿瘤生长显著减少。免疫组织化学分析显示巨噬细胞浸润在B16/ssIL-1 β中是强的，在B16/IL-1 β中是中等的，在对照肿瘤中是最小的。此外，在B16/ssIL-1 β肿瘤中检测到CD 4（+）细胞和分散的树突状细胞的中度浸润，而在B16/IL-1 β或对照肿瘤中观察到非常少或没有CD 4（+）细胞和树突状细胞。在体内生长后，所有肿瘤在其细胞表面上调ICAM-1。然而，与对照组相比，B16/ssIL-1 β组ICAM-1表达细胞的百分比高2 - 4倍。这些数据表明，IL-1 β在体内直接或间接地作为单核细胞、T辅助细胞和树突状细胞的趋化因子。这支持IL-1 β在肿瘤区域局部释放时对肿瘤生长具有调节作用。
Interleukin(IL)-1 differs from most other cytokines in its lack of a signal sequence. This results in intracellular retention of the immature preform. The release of IL-1 has been shown to be restricted predominantly to activated monocytes and macrophages and to be associated with apoptosis of the producer cell. These features have limited the investigation of IL-1 in early immune responses. In order to study the biological effects of local IL-1 beta release during an antitumour immune response, we used B16 mouse melanoma cells transduced with mature human IL-1 beta cDNA constructs. To obtain a released form of human IL-1 beta (ssIL-1 beta), the signal sequence from the related IL-1 receptor antagonist was ligated to the cDNA that encoded the mature form of IL-1 beta. When cells of the poorly immunogenic B16 melanoma cell line were transduced with IL-1 beta by retroviral infection, high levels of the protein were detected intracellularly, whereas cells transduced with IL-1 beta containing the signal sequence secreted most of their protein. The in vitro growth of the melanoma cells was unaffected by the IL-1 beta or ssIL-1 beta gene transfer. In contrast, the in vivo subcutaneous tumour growth of the ssIL-1 beta-transduced B16 cells in syngeneic C57BL/6 mice was significantly reduced compared with the IL-1 beta- and the mock-transduced controls. Immunohistochemical analysis revealed the infiltration of macrophages to be strong in B16/ssIL-1 beta, moderate in B16/IL-1 beta and minimal in control tumours. Furthermore, a moderate infiltration of CD4(+) cells and of scattered dendritic cells was detected in B16/ssIL-1 beta rumours whereas very few or no CD4(+) cells and dendritic cells were seen in the B16/IL-1 beta or control tumours. Following in vivo growth, all the tumours upregulated ICAM-1 on their cell surfaces. However, the percentage of ICAM-1-expressing cells was two- to four-fold higher in B16/ssIL-1 beta rumours compared ro the control. The data suggest that IL-1 beta acts in vivo, either directly or indirectly, as a chemotactic factor for monocytes, T helper cells and dendritic cells. This supports IL-1 beta having a regulatory effect on tumour growth when locally released in the tumour area.