Wu Chuan-ging, Budhu Anuradha, Chen Sheng, Zhou Xiaoling, Popescu Nicholas C, Valerie Kristoffer, Wang Xin Wei
Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892-4255, USA.
Mol Med. 2006 Jan-Mar;12(1-3):47-53. doi: 10.2119/2006-00020.Wu.
Hepatitis C virus (HCV) core protein features many intriguing properties and plays a pivotal role in cellular immunity, cell growth, apoptosis, cell transformation, and eventually in tumor development. However, the role of B cells, the primary players in the humoral immune response, during HCV infection is largely unknown. To explore the molecular effects of HCV core on human B cells, we conducted gene expression profiling of serial RNA samples from B cells that were infected with adenovirus harboring full-length HCV core protein and beta-galactosidase as a reference using a microarray platform containing 22,149 human oligo probes. The entire experiment was performed in duplicate in B lymphocytes that were isolated from two individual donors and incubated for up to 3 days after infection with adenovirus expressing HCV core protein to identify dynamic gene expression patterns. Differential expression of representative genes was validated by quantitative RT-PCR. We found that HCV core significantly inhibited B-lymphocyte apoptosis. We showed a dramatic downregulation of MHC class II molecules in B cells expressing HCV core, whereas the expression of immunoglobulin genes was not significantly altered. Moreover, genes associated with leukemia and B-lymphoma were consistently upregulated by HCV core. In contrast, downregulation of caspase-1 and caspase-4 was found to be associated with core's ability to prevent B-lymphocyte apoptosis. In summary, we have identified several clusters of genes that are differentially expressed in human B lymphocytes expressing HCV core, suggesting a potential impairment of antigen processing and presentation, which may provide more insights into HCV infection in B lymphocytes.
丙型肝炎病毒(HCV)核心蛋白具有许多引人关注的特性,在细胞免疫、细胞生长、细胞凋亡、细胞转化以及最终的肿瘤发展过程中发挥着关键作用。然而,作为体液免疫反应主要参与者的B细胞在HCV感染过程中的作用在很大程度上尚不清楚。为了探究HCV核心蛋白对人B细胞的分子影响,我们使用包含22,149个人寡核苷酸探针的微阵列平台,对感染携带全长HCV核心蛋白的腺病毒和作为参照的β-半乳糖苷酶的B细胞的系列RNA样本进行了基因表达谱分析。整个实验在从两名个体供体分离的B淋巴细胞中重复进行,在用表达HCV核心蛋白的腺病毒感染后孵育长达3天,以确定动态基因表达模式。代表性基因的差异表达通过定量逆转录-聚合酶链反应(qRT-PCR)进行验证。我们发现HCV核心蛋白显著抑制B淋巴细胞凋亡。我们发现表达HCV核心蛋白的B细胞中MHC II类分子显著下调,而免疫球蛋白基因的表达没有明显改变。此外,与白血病和B淋巴瘤相关的基因被HCV核心蛋白持续上调。相反,半胱天冬酶-1和半胱天冬酶-4的下调被发现与核心蛋白预防B淋巴细胞凋亡的能力有关。总之,我们已经鉴定出在表达HCV核心蛋白的人B淋巴细胞中差异表达的几个基因簇群,这表明抗原加工和呈递可能存在潜在损害,这可能为深入了解B淋巴细胞中的HCV感染提供更多见解。