Department of Dermatology, Henry Ford Hospital, Detroit, MI, USA.
J Dermatol Sci. 2010 Oct;60(1):8-20. doi: 10.1016/j.jdermsci.2010.07.007. Epub 2010 Jul 22.
Dysfunctions in memory T cells contribute to various inflammatory autoimmune diseases and neoplasms. We hypothesize that investigating the differences of genetic profiles between resting and activated naïve and memory T cells may provide insight into the characterization of abnormal memory T cells in diseases, such as Sézary syndrome (SS), a neoplasm composed of CD4(+) CD45RO(+) cells.
We determined genes distinctively expressed between resting and activated naive and memory cells. Levels of up-regulated genes in resting and activated memory cells were measured in SS PBMCs, which were largely comprised of CD4(+) CD45RO(+) cells, to quantitatively assess how different Sézary cells were from memory cells.
We compared gene expression profiles using high-density oligo-microarrays between resting and activated naïve and memory CD4(+) T cells. Differentially expressed genes were confirmed by qRT-PCR and immunoblotting. Levels of genes up-regulated in activated and resting memory T cells were determined in SS PBMCs by qRT-PCR.
Activated memory cells expressed greater numbers of immune-mediated genes involved in effector function compared to naïve cells in our microarray analysis and qRT-PCR. Nine out of 14 genes with enhanced levels in activated memory cells had reduced levels in SS PBMCs (p<0.05).
Activation of memory and naïve CD4(+) T cells revealed a diverging gap in gene expression between these subsets, with memory cells expressing immune-related genes important for effector function. Many of these genes were markedly depressed in SS patients, implying Sézary cells are markedly impaired in mounting immune responses compared to memory cells.
记忆 T 细胞功能障碍与各种炎症性自身免疫性疾病和肿瘤有关。我们假设，研究静止和激活的初始和记忆 T 细胞之间遗传谱的差异，可能有助于了解疾病中异常记忆 T 细胞的特征，如 SS（由 CD4+CD45RO+细胞组成的肿瘤）。
我们确定了静止和激活的初始和记忆细胞之间表达差异的基因。测量了大量由 CD4+CD45RO+细胞组成的 SS PBMC 中静止和激活记忆细胞中上调基因的水平，以定量评估不同的 Sézary 细胞与记忆细胞的差异。
我们使用高密度寡聚微阵列比较了静止和激活的初始和记忆 CD4+T 细胞的基因表达谱。通过 qRT-PCR 和免疫印迹验证差异表达的基因。通过 qRT-PCR 确定了 SS PBMC 中激活和静止记忆 T 细胞中上调基因的水平。
在我们的微阵列分析和 qRT-PCR 中，与初始细胞相比，激活的记忆细胞表达了更多参与效应功能的免疫调节基因。在激活的记忆细胞中上调的 14 个基因中有 9 个在 SS PBMC 中的水平降低（p<0.05）。
激活记忆和初始 CD4+T 细胞揭示了这两个亚群之间基因表达的明显差异，记忆细胞表达了对效应功能很重要的免疫相关基因。这些基因中的许多在 SS 患者中明显下调，这意味着与记忆细胞相比，Sézary 细胞在免疫反应中受到严重抑制。
