Sharif Saeed, Nakatani Yoshio, Wise Lyn, Corbett Michael, Real Nicola C, Stuart Gabriella S, Lateef Zabeen, Krause Kurt, Mercer Andrew A, Fleming Stephen B
Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
PLoS One. 2016 Dec 9;11(12):e0168007. doi: 10.1371/journal.pone.0168007. eCollection 2016.
Bovine papular stomatitis virus (BPSV) is a Parapoxvirus that induces acute pustular skin lesions in cattle and is transmissible to humans. Previous studies have shown that BPSV encodes a distinctive chemokine-binding protein (CBP). Chemokines are critically involved in the trafficking of immune cells to sites of inflammation and infected tissue, suggesting that the CBP plays a role in immune evasion by preventing immune cells reaching sites of infection. We hypothesised that the BPSV-CBP binds a wide range of inflammatory chemokines particularly those involved in BPSV skin infection, and inhibits the recruitment of immune cells from the blood into inflamed skin. Molecular analysis of the purified protein revealed that the BPSV-CBP is a homodimeric polypeptide with a MW of 82.4 kDa whilst a comprehensive screen of inflammatory chemokines by surface plasmon resonance showed high-affinity binding to a range of chemokines within the CXC, CC and XC subfamilies. Structural analysis of BPSV-CBP, based on the crystal structure of orf virus CBP, provided a probable explanation for these chemokine specificities at a molecular level. Functional analysis of the BPSV-CBP using transwell migration assays demonstrated that it potently inhibited chemotaxis of murine neutrophils and monocytes in response to CXCL1, CXCL2 as well as CCL2, CCL3 and CCL5 chemokines. In order to examine the effects of CBP in vivo, we used murine skin models to determine its impact on inflammatory cell recruitment such as that observed during BPSV infection. Intradermal injection of BPSV-CBP blocked the influx of neutrophils and monocytes in murine skin in which inflammation was induced with lipopolysaccharide. Furthermore, intradermal injection of BPSV-CBP into injured skin, which more closely mimics BPSV lesions, delayed the influx of neutrophils and reduced the recruitment of MHC-II+ immune cells to the wound bed. Our findings suggest that the CBP could be important in pathogenesis of BPSV infections.
牛丘疹性口炎病毒(BPSV)是一种副痘病毒,可在牛身上引发急性脓疱性皮肤损伤,并可传染给人类。先前的研究表明,BPSV编码一种独特的趋化因子结合蛋白(CBP)。趋化因子在免疫细胞向炎症部位和感染组织的迁移过程中起关键作用,这表明CBP通过阻止免疫细胞到达感染部位而在免疫逃避中发挥作用。我们推测,BPSV-CBP可结合多种炎性趋化因子,尤其是那些与BPSV皮肤感染有关的趋化因子,并抑制免疫细胞从血液中募集到炎症皮肤中。对纯化蛋白的分子分析表明,BPSV-CBP是一种分子量为82.4 kDa的同二聚体多肽,而通过表面等离子体共振对炎性趋化因子进行的全面筛选显示,它与CXC、CC和XC亚家族中的一系列趋化因子具有高亲和力结合。基于orf病毒CBP的晶体结构对BPSV-CBP进行的结构分析,在分子水平上为这些趋化因子特异性提供了一种可能的解释。使用Transwell迁移试验对BPSV-CBP进行的功能分析表明,它能有效抑制小鼠中性粒细胞和单核细胞对CXCL1、CXCL2以及CCL2、CCL3和CCL5趋化因子的趋化作用。为了研究CBP在体内的作用,我们使用小鼠皮肤模型来确定其对炎症细胞募集的影响,例如在BPSV感染期间观察到的情况。皮内注射BPSV-CBP可阻断脂多糖诱导炎症的小鼠皮肤中中性粒细胞和单核细胞的流入。此外,将BPSV-CBP皮内注射到更接近模拟BPSV损伤的受伤皮肤中,可延迟中性粒细胞的流入,并减少MHC-II+免疫细胞向伤口床的募集。我们的研究结果表明,CBP可能在BPSV感染的发病机制中起重要作用。