LABORATORY PROTOCOLS FOR THE QUANTITATION OF CYTOKINES BY BIOASSAY USING CYTOKINE RESPONSIVE CELL-LINES

Cytokines have been shown to be involved in many physiological process es, including the maintenance and control of a competent haematopoieti c/immune system. Abnormal cytokine secretion or synthesis can cause a wide range of pathological disorders. Cytokines have also been shown t o have therapeutic potential in many diseases. In order for the role o f cytokines to be evaluated in normal and pathogenic processes, it is vital that appropriate assay systems are used to measure their levels in different biological fluids,Whilst immunoassays may be a more conve nient method for quantitating cytokines, they only measure immunologic ally reactive material. They may or may not detect biologically inacti ve material, such as cytokines bound to soluble receptors or degraded cytokine molecules. Bioassays, however, detect biologically active cyt okines and can be as accurate and precise as immunoassays. The purpose of these protocols is to provide practical stepwise methods for the b ioassay of cytokines using cytokine responsive cell lines. They includ e tables of the most useful currently available cell Lines for the det ection of cytokines and how to maintain them. In addition, these proto cols provide all the materials and methods in a logical step-by-step p rocedure to carry out bioassays. Information is provided on possible p itfalls and general problems associated with bioassays and how to over come them. These protocols should be valuable to scientists new to the cytokine field as well as experienced scientists who require a consen sus methodology and access to information on cell lines useful for cyt okine bioassays.