Department of Rheumatology, Infectious Diseases and Laboratory Medicine, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
Department of Life Medical, Kyushu University of Health and Welfare, 1714-1 Yoshino, Nobeoka, Miyazaki, 882-8508, Japan.
Hum Cell. 2019 Apr;32(2):185-192. doi: 10.1007/s13577-018-00231-3. Epub 2018 Dec 17.
We previously reported the diversity of structure and integration sites of human T-cell leukemia virus type 1 (HTLV-1) provirus among different MT-2 cell lines. This raised the question as to whether cell phenotypes also differed among MT-2 cell lines. The influence of two different MT-2 cell lines (MT-2J and MT-2B) on the growth of the promonocytic leukemia cell line, U937, was investigated. Protein levels and mRNA expression of cytokines were also investigated. In addition, Western blot analysis of HTLV-1 regulatory proteins, Tax and HBZ, was also performed. Culture supernatant from MT-2B, but not MT-2J, cells showed marked suppressive effects on U937 cell growth. MT-2B showed high tumor necrosis factor (TNF)-α, TNF-β, and interferon (IFN)-γ both in protein levels of the culture supernatant and mRNA levels of the cells. Analysis using recombinant cytokines indicated that the suppressive effects of MT-2B were due, at least in part, to high levels of TNF-β and its synergic effects with IFN-γ in the culture supernatant. Protein levels of HTLV-1 Tax and HBZ were higher in MT-2B than those in MT-2J cells. These molecules have been reported to affect the cytokine production of HTLV-1 infected cells; therefore, the difference in these molecules may have accounted for the differences in cytokine production between MT-2J and MT-2B cells. Furthermore, because MT-2 cells showed a large variation of integrated HTLV-1 proviruses as well as cell phenotypes, it is important to exercise caution in the assessment and interpretation of experimental data from MT-2 cells.
我们之前曾报道过人 T 细胞白血病病毒 1(HTLV-1)前病毒在不同 MT-2 细胞系中的结构和整合位点多样性。这就提出了一个问题,即 MT-2 细胞系之间的细胞表型是否也存在差异。本研究调查了两种不同的 MT-2 细胞系(MT-2J 和 MT-2B)对 promonocytic 白血病细胞系 U937 生长的影响。还研究了细胞因子的蛋白水平和 mRNA 表达。此外,还进行了 HTLV-1 调节蛋白 Tax 和 HBZ 的 Western blot 分析。来自 MT-2B 而非 MT-2J 的细胞培养上清液对 U937 细胞生长具有明显的抑制作用。MT-2B 细胞培养上清液和细胞中 TNF-α、TNF-β 和 IFN-γ 的蛋白水平和 mRNA 水平均较高。使用重组细胞因子的分析表明,MT-2B 的抑制作用至少部分归因于培养上清液中 TNF-β 水平较高及其与 IFN-γ 的协同作用。MT-2B 细胞中 HTLV-1 Tax 和 HBZ 的蛋白水平高于 MT-2J 细胞。这些分子已被报道会影响 HTLV-1 感染细胞的细胞因子产生;因此,这些分子的差异可能导致了 MT-2J 和 MT-2B 细胞之间细胞因子产生的差异。此外,由于 MT-2 细胞表现出整合的 HTLV-1 前病毒和细胞表型的巨大变化,因此在评估和解释 MT-2 细胞的实验数据时需要谨慎。