Measurement of cytokines in clinical samples using immunoassays: problems and pitfalls

With the explosion in interest in cytokines and an increasing understanding of their possible involvement in the pathogenesis of many diseases, the number of studies measuring cytokines in body fluids has increased markedly over the last 5 to 10 years. However, it is readily apparent from the literature that the level of awareness of the many factors that can potentially influence the results obtained is very low. Such factors are likely to contribute considerably to the disparities seen among similar types of study. These include assay properties such as standardization and specificity, type of sample and sample handling, and many biological influences such as age, sex, ethnic background, and diurnal rhythmicity. Clearly, it is not possible to control for all these factors, but a minimum evaluation of assays should be carried out as described, including recovery and parallelity studies using relevant samples and the establishment of reproducibility and normal healthy population levels. An awareness of these factors, which is the purpose of this review, may hopefully lead to an improvement in the quality of studies incorporating the measurement of cytokines in clinical samples and should aid in the interpretation of the data.