Chi Jiaqi, You Leiming, Li Peipei, Teng Man, Zhang Gaiping, Luo Jun, Wang Aiping
Department of Oncology, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing, 100078, People's Republic of China.
School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 100029, People's Republic of China.
Virus Genes. 2018 Apr;54(2):236-245. doi: 10.1007/s11262-018-1535-6. Epub 2018 Jan 25.
Infectious bursal disease virus (IBDV) is an important immunosuppressive virus in chickens. Surface immunoglobulin M (sIgM)-bearing B lymphocytes act as the major targets of IBDV in the bursa of Fabricius, and sIgM may function as one of the membrane binding sites responsible for IBDV infection. Recently, using the virus overlay protein binding assay, the chicken λ light chain of sIgM was identified to specifically interact with IBDV in a virulence-independent manner in vitro. To further investigate sIgM λ light chain-mediated IBDV binding and infection in pre-B cells, the cell line DT40, which is susceptible to both pathogenic and attenuated IBDV, was used. Based on the RNA interference strategy, the DT40 cell line whose λ light chain of sIgM was stably knocked down, herein termed DT40LKD, was generated by the genomic integration of a specific small hairpin RNA and a green fluorescence protein co-expression construct. Flow cytometry analysis indicated that the binding of IBDV to DT40LKD cells was significantly reduced due to the loss of sIgM λ light chain. In particular, reduced viral replication was observed in IBDV-incubated DT40LKD cells, and no viral release into cell culture medium was detected by the IBDV rapid diagnostic strips. In addition, the rescue of sIgM λ light chain expression restored viral binding and replication in DT40LKD cells. These results show that sIgM λ light chain appears to be beneficial for IBDV attachment and infection, suggesting that sIgM acts as a binding site involved in IBDV infection.
传染性法氏囊病病毒(IBDV)是鸡体内一种重要的免疫抑制病毒。带有表面免疫球蛋白M(sIgM)的B淋巴细胞是IBDV在法氏囊中的主要靶细胞,sIgM可能作为负责IBDV感染的膜结合位点之一发挥作用。最近,通过病毒覆盖蛋白结合试验,在体外鉴定出sIgM的鸡λ轻链以毒力非依赖的方式与IBDV特异性相互作用。为了进一步研究sIgM λ轻链介导的IBDV在pre - B细胞中的结合和感染,使用了对致病性和减毒IBDV均易感的DT40细胞系。基于RNA干扰策略,通过特定小发夹RNA与绿色荧光蛋白共表达构建体的基因组整合,构建了sIgM λ轻链稳定敲低的DT40细胞系,在此称为DT40LKD。流式细胞术分析表明,由于sIgM λ轻链的缺失,IBDV与DT40LKD细胞的结合显著减少。特别是,在IBDV孵育的DT40LKD细胞中观察到病毒复制减少,并且通过IBDV快速诊断试纸条未检测到病毒释放到细胞培养基中。此外,sIgM λ轻链表达的恢复恢复了DT40LKD细胞中的病毒结合和复制。这些结果表明,sIgM λ轻链似乎有利于IBDV的附着和感染,提示sIgM作为参与IBDV感染的结合位点发挥作用。