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

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.

► Analysis of the effect of preanalytic factors on IL-16 quantitation is missing as yet.
► For IL-16 measurement only sera or cell-free plasma samples should be used.
► Pro-IL-16 represents the major IL-16 form in cell-rich plasma samples.
► Commercially IL-16 ELISA are not specific for mature C-IL-16.
► New methods are necessary for quantitation of C-IL-16 and pro-IL-16 uniquely.