Methodologic issues in the measurement of interleukin-16 in clinical blood samples using immunoassays

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• 作者：Alexander Goihl^a ; ¹ ; Anna-Maria Rolle^a ; ¹ ; Thilo Kä ; hne^b ; Annegret Reinhold^a ; Sabine Wrenger^a ; Dirk Reinhold^a ; ^{dirk.reinhold@med.ovgu.de}
• 关键词：C-IL-16 ; C-terminal IL-16 form ; ELISA ; enzyme-linked immunosorbent assay ; IL-16 ; interleukin-16 ; IP ; immunoprecipitation ; N-IL-16 ; N-terminal IL-16 form ; Pro-IL-16 ; IL-16 precursor protein ; RA ; rheumatoid arthritis ; SLE ; systemic lupus erythematosus
• 刊名：Cytokine
• 出版年：2012
• 期刊代码：97_10434666
• 类别：imm
• 出版时间：April, 2012
• 卷：58
• 期：1
• 页码：1-5
• 文件大小：446 K

摘要

Quantitation of interleukin-16 (IL-16) in clinical blood samples has strongly increased, since IL-16 appears to be involved in the pathogenesis of several inflammatory diseases. IL-16 is synthesized in the cell cytoplasm as precursor protein (pro-IL-16), which can be processed by caspase-3 into N-terminal (N-IL-16) and C-terminal (C-IL-16) fragments. C-IL-16 is described to be subsequently secreted. Using commercially available IL-16 ELISA, a pro-IL-16 ELISA and immunoprecipitation analysis, we investigated, whether type and handling of blood samples influence IL-16 quantitation and whether existing IL-16 ELISA are specific for C-IL-16. We observed that cell-rich plasma samples reflect falsely-elevated IL-16 concentrations due to cell contaminations. Interestingly, not C-IL-16, but pro-IL-16 represents the major IL-16 form in cell-rich plasma samples. Notably, commercially IL-16 ELISA could not distinguish between C-IL-16 and pro-IL-16. Thus, cell-rich plasma samples should not be used for IL-16 measurements and new methods are necessary for quantitation of C-IL-16 and pro-IL-16 uniquely.