Development of an 8-plex Luminex assay to detect swine cytokines for vaccine development: Assessment of immunity after porcine reproductive and respiratory syndrome virus (PRRSV) vaccination
Development of an 8-plex Luminex assay to detect swine cytokines for vaccine development: Assessment of immunity after porcine reproductive and respiratory syndrome virus (PRRSV) vaccination
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DOI：
10.1016/j.vaccine.2010.05.016
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发表时间：
2010-07-19
期刊：
影响因子：
5.5
通讯作者：
Christopher-Hennings, Jane
中科院分区：
文献类型：
--
作者：
Lawson, Steven;Lunney, Joan;Christopher-Hennings, Jane
Luminex（Luminex Corp.，奥斯汀，德克萨斯州）开发了多重猪细胞因子测定法，可同时测量猪血清中的8种细胞因子，用于评估候选疫苗。在猪繁殖与呼吸综合征病毒（PRRSV）疫苗/攻毒研究中，对存档血清进行荧光微球免疫测定（FMIA）。该FMIA同时检测先天性（IL-1 β、IL-8、IFN-α、TNF-α、IL-12）、调节性（IL-10）、Th 1（IFN-γ）和Th 2（IL-4）细胞因子。测量这些蛋白质以评估与提供针对PRRSV的不同水平的保护性免疫的疫苗接种策略相关的血清细胞因子水平。猪用改良活病毒（MLV）疫苗接种，随后用不相同的PRRSV分离株（糖蛋白（GP 5）基因具有93%的同一性）攻毒。在PRRSV攻毒后，在MLV接种猪中观察到保护（定义为无血清病毒血症），但在接种含佐剂灭活病毒疫苗（KV/ADJ）（GP 5基因与攻毒株的同一性> 99%）或未接种的猪中未观察到保护。与MLV接种组和对照组相比，在KV/ADJ组中观察到IL-12水平显著升高。然而，血清IL-12的这种显著增加与针对PRRSV病毒血症的保护无关。使用该测定来测量局部细胞因子组织反应的其他研究可能有助于定义保护性细胞因子反应，并且将有助于有效疫苗的靶向设计，不仅用于PRRSV，而且用于其他猪病原体。(C)2010爱思唯尔有限公司版权所有。
A Luminex (Luminex Corp., Austin, TX) multiplex swine cytokine assay was developed to measure 8 cytokines simultaneously in pig serum for use in assessment of vaccine candidates. The fluorescent microsphere immunoassay (FMIA) was tested on archived sera in a porcine reproductive and respiratory syndrome virus (PRRSV) vaccine/challenge study. This FMIA simultaneously detects innate (IL-1 beta, IL-8, IFN-alpha, TNF-alpha, IL-12), regulatory (IL-10), Th1 (IFN-gamma) and Th2 (IL-4) cytokines. These proteins were measured to evaluate serum cytokine levels associated with vaccination strategies that provided for different levels of protective immunity against PRRSV. Pigs were vaccinated with a modified-live virus (MLV) vaccine and subsequently challenged with a non-identical PRRSV isolate (93% identity in the glycoprotein (GP5) gene). Protection (as defined by no serum viremia) was observed in the MLV vaccinated pigs after PRRSV challenge but not those vaccinated with killed virus vaccine with adjuvant (KV/ADJ) (99% identity in the GP5 gene to the challenge strain) or non-vaccinates. Significantly elevated levels of IL-12 were observed in the KV/ADJ group compared to MLV vaccinated and control groups. However, this significant increase in serum IL-12 did not correlate with protection against PRRSV viremia. Additional studies using this assay to measure the local cytokine tissue responses may help in defining a protective cytokine response and would be useful for the targeted design of efficacious vaccines, not only for PRRSV, but also for other swine pathogens. (C) 2010 Elsevier Ltd. All rights reserved.