The effect of HTLV-1 bZIP factor (HBZ) RNA and protein in mouse CD4 T cells

Human T-cell leukemia virus type 1 (HTLV-1) bZIP factor (HBZ) which is encoded in the minus strand of HTLV-1 provirus, possesses dual function as protein, and also as RNA. To know the effect of HBZ RNA and protein in primary T-cells, we introduced HBZ, or its mutant TTG (exclude protein activity), or SM (exclude RNA activity) with retrovirus vector into CD4 positive murine T cells, and analysed the transcriptome profiling. We found that HBZ RNA altered cell cycle progression, cell survival related genes, while HBZ protein altered immunology related genes. This microarray results demonstrated that HBZ RNA and protein possess distinct functions in primary cells. CD4 positive cells were enriched from murine splenocyte, and cultured with irradiated antigen presenting cells (APC). HBZ, or its mutant were introduced with retroviral vector. Forty eight hours after introduction, cells were washed and cultured with recombinant human IL-2. Further 48 hours after culture, virus infected cells were sorted and analysed by agilent microarray.

人类T细胞白血病病毒1型（HTLV-1）编码的bZIP因子（HBZ）位于HTLV-1前病毒的负链，同时可分别以蛋白质和RNA两种形式行使功能。为探究HBZ RNA与HBZ蛋白在原代T细胞中的生物学效应，我们通过逆转录病毒载体将HBZ、其突变体TTG（丧失蛋白活性）或SM（丧失RNA活性）导入CD4阳性小鼠原代T细胞，并开展转录组表达谱分析。结果显示，HBZ RNA可调控细胞周期进程与细胞存活相关基因的表达，而HBZ蛋白则主要影响免疫相关基因的转录水平。本次微阵列实验结果证实，HBZ RNA与HBZ蛋白在原代细胞中具备截然不同的功能。我们从小鼠脾脏淋巴细胞中分离富集CD4阳性T细胞，将其与辐照处理后的抗原呈递细胞（APC）共培养，随后通过逆转录病毒载体导入HBZ或其对应突变体。转导48小时后，清洗细胞并更换为含重组人白细胞介素2（IL-2）的培养基进行培养。继续培养48小时后，分选病毒感染的阳性细胞，采用安捷伦（Agilent）芯片完成转录组分析。