Zoon C K, Seitelman E, Keller S, Graham L, Blevins T L, Dumur C I, Bear H D
Department of Surgery, Virginia Commonwealth University Health System, Richmond, VA, USA.
Division of Surgical Oncology, Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.
Cancer Gene Ther. 2014 Oct;21(10):441-7. doi: 10.1038/cgt.2014.48. Epub 2014 Sep 19.
We have previously demonstrated that expansion of activated tumor-sensitized T cells in interleukin (IL)-7/15 results in greater expansion and antitumor activity than expansion in IL-2. We sought to determine whether T cells exposed to IL-2 versus IL-7/15 exhibited distinct gene expression patterns. Lymphocytes were harvested from Pmel-1 mice immunized with B16-GMCSF melanoma cells, activated in vitro, and cultured in IL-2 or IL-7/15 for 1, 3 or 6 days. T cells were harvested and analyzed using microarray, real-time quantitative polymerase chain reaction (RT-QPCR) or sorted into T-cell subsets and analyzed. We found significant differences in gene expression for T cells cultured in IL-2 versus IL-7/15, starting on day 3. This was not a function of subset differentiation; when T cells were divided into subsets, the central memory (T(CM)), effector memory (T(EM)) and effector (T(E)) T cells cultured in the IL-2 more closely resembled each other than the identical phenotypic subset exposed to IL-7/15. Thus, the differences in gene expression induced by culture in IL-2 versus IL-7/15 do not merely reflect differences in the frequency of T(CM) versus T(EM) versus T(E) cells, but rather reflect that the gene expression levels of those T-cell subsets when exposed to different cytokines are fundamentally different.
我们之前已经证明,与在白细胞介素(IL)-2中扩增相比,在IL-7/15中扩增活化的肿瘤致敏T细胞会导致更大程度的扩增和抗肿瘤活性。我们试图确定暴露于IL-2与IL-7/15的T细胞是否表现出不同的基因表达模式。从用B16-GMCSF黑色素瘤细胞免疫的Pmel-1小鼠中收获淋巴细胞,在体外活化,并在IL-2或IL-7/15中培养1、3或6天。收获T细胞并使用微阵列、实时定量聚合酶链反应(RT-QPCR)进行分析,或将其分选到T细胞亚群中并进行分析。我们发现,从第3天开始,在IL-2与IL-7/15中培养的T细胞在基因表达上存在显著差异。这不是亚群分化的作用;当T细胞被分为亚群时,在IL-2中培养的中央记忆(T(CM))、效应记忆(T(EM))和效应(T(E))T细胞彼此之间的相似性高于暴露于IL-7/15的相同表型亚群。因此,在IL-2与IL-7/15中培养所诱导的基因表达差异不仅仅反映了T(CM)、T(EM)与T(E)细胞频率的差异,而是反映出这些T细胞亚群在暴露于不同细胞因子时的基因表达水平存在根本差异。