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• W360646757 abstract "Poxviruses are large DNA viruses that replicate in the cytoplasm, and encode numerous host immune modulating factors. The parapoxvirus, orf virus (ORFV) is a zoonotic pathogen that causes pustular dermatitis in sheep, goats and humans. The fact that ORFV can repeatedly infect its host may be explained by the discovery of several secreted immunomodulators. One of the secreted viral proteins is a chemokine binding protein (CBP) that shares low homology with the type II poxvirus CBPs. These viral CBPs have no known mammalian homologue and are unique in their ability to specifically bind CC chemokines with high affinity. The inflammatory CC chemokines play a role in recruiting monocytes and dendritic cells to the site of infection during innate immunity, and constitutive CC chemokines are required for the migration of dendritic cells to the lymph nodes to initiate the adaptive immune response. This study examined the ability of ORFV CBP to block chemokine gradients formed during innate and adaptive immune responses.In this study, the ability of ORFV CBP to bind murine inflammatory and constitutive CC chemokines was assessed using murine models. The immune responses in mice are well characterised and murine reagents are readily available allowing detailed analysis of both in vitro and in vivo studies. CBP bound murine chemokines CCL2, CCL3, CCL5, CCL19, and CCL21 with high affinity by surface plasmon resonance studies. The inflammatory chemokines CCL2, CCL3 and CCL5 are produced during damage and inflammation in epithelial tissues, and are involved in recruiting antigen-presenting cells to the site of acute inflammation. Transwell migration assays were used to determine optimal amounts of these chemokines required to induce migration of monocytes and immature dendritic cells. Monocytes and dendritic cells migrated in maximal numbers in response to between 12.5 – 50 ng/ml of inflammatory chemokines. In the presence of increasing amounts of CBP, the chemokine-induced migration of monocytes and dendritic cells was inhibited in a dose dependent manner. A molar ratio of chemokine to CBP of 1:4 reduced the CCL2-, CCL3-, and CCL5-induced migration of monocytes and immature dendritic cells to background levels. The chemotactic ability of CCL19 and CCL21 for mature dendritic cells was also assessed using transwell migration assays. CCL19 and CCL21 were used at 12.5 ng/ml to induce maximal migration of mature dendritic cells. A molar ratio of chemokine to CBP of 1:4 completely inhibited CCL19-induced migration of mature dendritic cells. CBP bound murine CCL21 with comparatively lower binding affinity and hence a higher chemokine to CBP ratio of 1:32 was required to reduce chemotaxis to background levels. Further functional assays utilized murine models to study the effects of CBP on cellular trafficking in vivo.The administration of lipopolysaccharide (LPS) in skin results in a highly localised inflammation with increased expression of the inflammatory chemokines CCL2, CCL3, and CCL5. The ability of CBP to bind these…" @default.
• W360646757 created "2016-06-24" @default.
• W360646757 creator A5079009208 @default.
• W360646757 date "2011-01-01" @default.
• W360646757 modified "2023-09-28" @default.
• W360646757 title "Functional characterisation of orf virus chemokine binding protein" @default.
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