Methods for Study of Chemokine Receptors in the Tissues

Three different assays were used to study the distribution of binding sites for IL-8 in human skin and several animal tissues. An in situ binding assay was designed in which the binding of radiolabeled IL-8 to small intact tissue pieces was studied, and a histological autoradiographic technique was used to detect the bound chemokine in the subsequently prepared tissue sections. A modified assay was also performed in which the binding of unlabeled IL-8 to intact tissue pieces was visualized using monoclonal anti-IL-8 antibody. In addition, we performed a "classical" autoradiographic study in which radiolabeled IL-8 was injected subcutaneously and visualized in sections prepared from the injected sites by autoradiography. We reflect on the potentials and limitations of studying the chemokine binding in situ, compare the results, and discuss the relative advantages and disadvantages of each of the techniques used.