Thymic Epithelial Cells with Human T cell Leukemia Virus type 1 (HTLV-1) Infected T cell supernatant (CEM, CIB, C91PL)

The human T-lymphotropic virus type-1 (HTLV-1) is the cause of adult T cell leukemia/lymphoma (ATL) and HTLV-1 associated myelopathy/tropical spatic paraparesis (HAM/TSP). Both diseases have a late onset, although ATL has a medium survivor of 7.7 months after the onset despise aggressive treatment. T CD4+ cells are the main target of HTLV-1, but other cells types are known to be infected as T CD8+, CD34+ progenitor, dendritic cells and immature lymphocytes. The thymus gland is a primary lymphoid organ, in which the lymphocytes undergo differentiation, where selected T cell ultimately being exported from the organ and going to peripheral lymphoid organs. This process occurs along with immature lymphocyte migration and interaction with thymic microenvironment. Thymic epithelial cells (TECs) are the main component of the thymic stroma and are responsible for the process of intrathymic T cell maturation. This process are dependent of T cell receptor (TCR)-mediated recognition of antigenic peptide fragments presented by major histocompatibility complex (MHC) molecules in TEC and culminate in TCR repertoire formation. In this study, we show that TECs have the receptors for HTLV-1 entry and can be infected by cell-cell contact and cell-free virus. These cells change gene expression of anti-apoptosis, chemokine and adhesion molecules genes; however, there is no difference in antigen presentation molecules. Furthermore, HTLV-1 infected TECs can transmit the virus to a CD4 T cell lineage and CD4 T cells derived from peripheral blood of healthy donors after in vitro co-cultivation. Conjointly, our data points to the possibility that the human thymic epithelial cells must favor the HTLV-1 infection as a reservoir, transmitting the virus to maturing lymphocytes that cause the disease in the periphery. We used thymic epithelial cell (TEC) treated with non-infected T cell supernatant (CEM) and TEC treated with infected T cell supernatant (CIB or C91PL). We analyze three independent samples (biological replicates) for each conditions (total 3 conditions) and also technical replicates using dye-swap. In total we use 4.5 slides with 4 array in each.

人类T细胞嗜淋巴细胞病毒1型（human T-lymphotropic virus type-1, HTLV-1）是成人T细胞白血病/淋巴瘤（adult T cell leukemia/lymphoma, ATL）以及HTLV-1相关性脊髓病/热带痉挛性截瘫（HTLV-1 associated myelopathy/tropical spastic paraparesis, HAM/TSP）的致病病原体。两种疾病均呈迟发性发病特征，其中成人T细胞白血病/淋巴瘤即便接受积极治疗，发病后的中位生存期仅为7.7个月。CD4+ T细胞是HTLV-1的主要靶细胞，但已有研究证实其他多种细胞类型也可被该病毒感染，包括CD8+ T细胞、CD34+造血祖细胞、树突状细胞以及未成熟淋巴细胞。胸腺作为一级淋巴器官，是淋巴细胞分化发育的核心场所：经筛选后的成熟T细胞最终迁出胸腺，迁移至外周淋巴器官。这一成熟过程伴随未成熟淋巴细胞的迁移，以及其与胸腺微环境的动态相互作用。胸腺上皮细胞（thymic epithelial cells, TECs）是胸腺基质的主要组成成分，负责调控胸腺内T细胞的成熟过程。该过程依赖于T细胞受体（T cell receptor, TCR）识别由胸腺上皮细胞表面主要组织相容性复合体（major histocompatibility complex, MHC）分子呈递的抗原肽片段，最终完成T细胞受体库的塑造。本研究证实，胸腺上皮细胞携带HTLV-1入侵受体，可通过细胞间接触以及无细胞病毒两种途径被感染。感染后，这些细胞的抗凋亡基因、趋化因子基因与黏附分子基因的表达谱发生显著改变，但抗原呈递分子的表达无明显差异。此外，体外共培养实验结果显示，HTLV-1感染的胸腺上皮细胞可将病毒传播至CD4 T细胞系，以及健康供者外周血来源的CD4 T细胞。综上，本研究数据提示，人类胸腺上皮细胞可作为HTLV-1的储存库促进病毒感染，并将病毒传递至成熟淋巴细胞，最终在外周组织引发相关疾病。本研究设置两组实验处理：分别采用未感染T细胞上清（CEM细胞株）处理的胸腺上皮细胞，以及感染HTLV-1的T细胞上清（CIB或C91PL细胞株）处理的胸腺上皮细胞。针对每种处理条件，我们设置3次独立生物学重复（总计3组实验条件），同时采用染料互换（dye-swap）方法设置技术重复。本研究总计使用4.5张芯片玻片，每张玻片搭载4张基因芯片阵列。