γ干扰素和α肿瘤坏死因子对培养的人肌肉细胞的协同细胞毒性作用。

Synergistic cytotoxic effect of interferon-gamma and tumor necrosis factor-alpha on cultured human muscle cells.

作者信息

Kalovidouris A E, Plotkin Z

机构信息

Rheumatology Section, Richard L. Roudebush Veterans Affairs Medical Center, Indianapolis, IN 46202, USA.

出版信息

J Rheumatol. 1995 Sep;22(9):1698-703.

PMID:8523348

Abstract

OBJECTIVE

To investigate the effects of human interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), either alone or in combination, on the viability of human muscle cells in culture.

METHODS

Cultures of human muscle cells were treated with various concentrations of recombinant IFN-gamma and TNF-alpha alone and in combination, and the cytotoxic effects of the cytokines on muscle cells were assessed by measuring lactic dehydrogenase (LDH) release in supernatants and by observation of the cells for morphologic changes under phase microscopy.

RESULTS

Exposure of muscle cells to 100 U/ml of either IFN-gamma or TNF-alpha for 9 days caused no cytotoxic effects, as assessed by LDH release in supernatants of muscle cell cultures and by microscopic observation of the cell cultures. However, when IFN-gamma and TNF-alpha were added together in the muscle cell cultures, they caused significant cytotoxic effects. Thus, in combination, IFN-gamma and TNF-alpha at 100 U/ml each caused significant release of LDH (3rd day 9%, 4th day 28.5%, 7th day 55.5%, 9th day 74%) in the supernatants of treated cultures compared to controls. Moreover, inspection by phase microscopy showed clear damage of muscle cells; from Days 3 to 4 progressive vacuolation, detachment of cells, and finally disintegration of the muscle cells by the 8th to 10th day was observed. The synergistic cytotoxic effect of IFN-gamma and TNF-alpha occurred at concentrations as low as 1 U/ml and 10 U/ml, respectively.

CONCLUSION

Our study demonstrates for the first time a direct synergistic cytotoxic effect of IFN-gamma and TNF-alpha on human muscle cells in culture. Given that T cells and macrophages are prominent in the chronic inflammatory cell infiltrates of the affected muscles in patients with myositis, our findings suggest that IFN-gamma and TNF-alpha may play an important role in the pathogenesis of muscle destruction of this disorder.

摘要

目的

研究人γ干扰素（IFN-γ）和肿瘤坏死因子-α（TNF-α）单独或联合使用对培养的人肌肉细胞活力的影响。

方法

用不同浓度的重组IFN-γ和TNF-α单独及联合处理人肌肉细胞培养物，通过测量上清液中乳酸脱氢酶（LDH）的释放以及在相差显微镜下观察细胞形态变化来评估细胞因子对肌肉细胞的细胞毒性作用。

结果

通过肌肉细胞培养上清液中LDH的释放以及细胞培养物的显微镜观察评估，肌肉细胞暴露于100 U/ml的IFN-γ或TNF-α 9天未产生细胞毒性作用。然而，当IFN-γ和TNF-α一起添加到肌肉细胞培养物中时，它们产生了显著的细胞毒性作用。因此，联合使用时，与对照组相比，100 U/ml的IFN-γ和TNF-α各自导致处理后培养物上清液中LDH显著释放（第3天9%，第4天28.5%，第7天55.5%，第9天74%）。此外，相差显微镜检查显示肌肉细胞明显受损；观察到从第3天到第4天逐渐出现空泡化、细胞脱落，到第8天至第10天肌肉细胞最终解体。IFN-γ和TNF-α的协同细胞毒性作用分别在低至1 U/ml和10 U/ml的浓度下出现。