Rechsteiner Markus P, Berger Christoph, Zauner Ludwig, Sigrist Jürg A, Weber Matthias, Longnecker Richard, Bernasconi Michele, Nadal David
Experimental Infectious Diseases and Cancer Research, Division of Infectious Diseases and Hospital Epidemiology, University Children's Hospital of Zurich, Zurich, Switzerland.
J Virol. 2008 Feb;82(4):1739-47. doi: 10.1128/JVI.01723-07. Epub 2007 Dec 5.
The B-lymphotropic Epstein-Barr virus (EBV) encodes two isoforms of latent membrane protein 2 (LMP2), LMP2A and LMP2B, which are expressed during latency in B cells. The function of LMP2B is largely unknown, whereas LMP2A blocks B-cell receptor (BCR) signaling transduction and induction of lytic EBV infection, thereby promoting B-cell survival. Transfection experiments on LMP2B in EBV-negative B cells and the silencing of LMP2B in EBV-harboring Burkitt's lymphoma-derived Akata cells suggest that LMP2B interferes with the function of LMP2A, but the role of LMP2B in the presence of functional EBV has not been established. Here, LMP2B, LMP2A, or both were overexpressed in EBV-harboring Akata cells to study the function of LMP2B. The overexpression of LMP2B increased the magnitude of EBV switching from its latent to its lytic form upon BCR cross-linking, as indicated by a more-enhanced upregulation and expression of EBV lytic genes and significantly increased production of transforming EBV compared to Akata vector control cells or LMP2A-overexpressing cells. Moreover, LMP2B lowered the degree of BCR cross-linking required to induce lytic EBV infection. Finally, LMP2B colocalized with LMP2A as demonstrated by immunoprecipitation and immunofluorescence and restored calcium mobilization upon BCR cross-linking, a signaling process inhibited by LMP2A. Thus, our findings suggest that LMP2B negatively regulates the function of LMP2A in preventing the switch from latent to lytic EBV replication.
嗜B淋巴细胞的爱泼斯坦-巴尔病毒(EBV)编码两种潜伏膜蛋白2(LMP2)的异构体,即LMP2A和LMP2B,它们在B细胞潜伏期间表达。LMP2B的功能在很大程度上尚不清楚,而LMP2A可阻断B细胞受体(BCR)信号转导和EBV裂解性感染的诱导,从而促进B细胞存活。对EBV阴性B细胞进行LMP2B转染实验以及对携带EBV的伯基特淋巴瘤来源的Akata细胞中的LMP2B进行沉默实验表明,LMP2B会干扰LMP2A的功能,但LMP2B在功能性EBV存在时的作用尚未明确。在此,我们在携带EBV的Akata细胞中过表达LMP2B、LMP2A或两者,以研究LMP2B的功能。与Akata载体对照细胞或过表达LMP2A的细胞相比,LMP2B的过表达增加了BCR交联后EBV从潜伏形式转变为裂解形式的程度,这表现为EBV裂解基因的上调和表达增强,以及转化型EBV的产生显著增加。此外,LMP2B降低了诱导EBV裂解性感染所需的BCR交联程度。最后,免疫沉淀和免疫荧光证明LMP2B与LMP2A共定位,并在BCR交联后恢复了钙动员,而这一信号传导过程受到LMP2A的抑制。因此,我们的研究结果表明,LMP2B在阻止EBV从潜伏复制转变为裂解复制方面对LMP2A的功能起负调节作用。