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Choice of immunoassay to evaluate porcine cytokine levels
Veterinary Immunology and Immunopathology ( IF 1.8 ) Pub Date : 2020-10-06 , DOI: 10.1016/j.vetimm.2020.110129
Nora L Ueland 1 , Judith K Ludvigsen 2 , Bernt C Hellerud 3 , Tom E Mollnes 4 , Espen W Skjeflo 5
Affiliation  

Background

In order to adequately monitor cytokines in experimental models, currently available methods and commercially available kits should be compared.

Aim

To compare the plasma and tissue concentrations of IL-1β, IL-6, IL-8, IL-10, and TNF as a measure of systemic inflammation in septic pigs.

Methods

Cytokines were quantified from blood and tissue samples obtained at 0, 60, 120, 180, and 240 min, and in postmortem biopsies of the liver, kidney, lung, heart, and spleen from 26 anesthetized landrace pigs. (24 with experimental sepsis, two sham controls). Porcine-specific ELISAs (R&D) and multiplex (9-plex from Thermo Fischer, 13-plex from Millipore) immunoassays were compared.

Results

The assays differed for the different cytokines and between blood and tissue. In blood, the highest concentration of TNF and IL-6 was in ELISA, IL-1β equal in ELISA and 13-plex, IL-8 in 13-plex and IL-10 in 9-plex. In tissue, the highest concentration of TNF and IL-1β was in ELISA, IL-6 and IL-8 in 13-plex and IL-10 in 9-plex.

Conclusion

The choice of analysis impacts the quantified cytokine responses in porcine models. ELISA and multiplex techniques supplement each other and our data suggest which assays to use for the quantification of the different cytokines.



中文翻译:

选择免疫测定法评估猪细胞因子水平

背景

为了充分监测实验模型中的细胞因子,应比较当前可用的方法和市售的试剂盒。

目标

为了比较IL-1β,IL-6,IL-8,IL-10和TNF的血浆和组织浓度,以作为化脓性猪全身性炎症的指标。

方法

从在0、60、120、180和240分钟获得的血液和组织样品中以及从26头麻醉的地方猪的肝,肾,肺,心脏和脾脏进行的活检中定量细胞因子。(24个实验性败血症,两个假对照)。比较了猪特异性ELISA(R&D)和多重免疫检测(Thermo Fischer的9重检测,密理博公司的13重检测)。

结果

对于不同的细胞因子以及血液和组织之间的测定方法不同。在血液中,TNF和IL-6的最高浓度在ELISA中,IL-1β在ELISA和13-plex中相等,IL-8在13-plex中,IL-10在9-plex中。在组织中,TNF和IL-1β的最高浓度在ELISA中,在13重复合物中为IL-6和IL-8,在9重复合物中为IL-10。

结论

分析的选择会影响猪模型中定量的细胞因子反应。ELISA和多重技术相辅相成,我们的数据表明可使用哪种测定法定量不同的细胞因子。

更新日期:2020-10-12
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