The challenge of measuring IL-33 in serum using commercial ELISA: lessons from asthma
The challenge of measuring IL-33 in serum using commercial ELISA: lessons from asthma
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DOI：
10.1111/cea.12718
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发表时间：
2016-06-01
影响因子：
6.1
通讯作者：
Sayers, I.
中科院分区：
文献类型：
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作者：
Ketelaar, M. E.;Nawijn, M. C.;Sayers, I.
背景白细胞介素-33（IL-33）在炎症性疾病尤其是哮喘中的作用已得到广泛研究。许多人的生物样本，包括血清，已被用来确定IL-33的蛋白水平，旨在研究其参与哮喘。研究IL-33与疾病的关系需要可靠的方法，特别是考虑到血清样品的复杂性。我们评估了四种IL-33 ELISA试剂盒，旨在确定一种稳健且可重复的方法来定量哮喘患者血清中的IL-33。（两个研发系统），ADI-900-201（Enzo Life Sciences）和SKR 038（GenWay Biotech Inc San Diego USA）进行免疫测定，以及使用重组IL-33及其可溶性受体IL-1 RL 1-a进行加标实验。2）人血清部分通过IL-1 RL 1-a干扰IL-33定量; 3）使用非血清认证的试剂盒可能会导致虚假的结果。由于IL-33/IL-1 RL 1在哮喘等疾病患者血清中的表达水平有待进一步研究，因此需要一种更灵敏、更特异的检测方法，这对于进一步了解和靶向IL-33/IL-1 RL 1轴在人类疾病中的作用至关重要。
BackgroundInterleukin-33 (IL-33) has been subject of extensive study in the context of inflammatory disorders, particularly in asthma. Many human biological samples, including serum, have been used to determine the protein levels of IL-33, aiming to investigate its involvement in asthma. Reliable methods are required to study the association of IL-33 with disease, especially considering the complex nature of serum samples.ObjectiveWe evaluated four IL-33 ELISA kits, aiming to determine a robust and reproducible approach to quantifying IL-33 in human serum from asthma patients.MethodsIL-33 levels were investigated in serum of well-defined asthma patients by the Quantikine, DuoSet (both R&D systems), ADI-900-201 (Enzo Life Sciences), and SKR038 (GenWay Biotech Inc San Diego USA) immunoassays, as well as spiking experiments were performed using recombinant IL-33 and its soluble receptor IL-1RL1-a.ResultsWe show that 1) IL-33 is difficult to detect by ELISA in human serum, due to lack of sensitivity and specificity of currently available assays; 2) human serum interferes with IL-33 quantification, in part through IL-1RL1-a; and 3) using non-serum certified kits may lead to spurious findings.Conclusion and Clinical RelevanceIf IL-33 is to be studied in the serum of asthma patients and other diseases, a more sensitive and specific assay method is required, which will be vital for further understanding and targeting of the IL-33/IL-1RL1 axis in human disease.