Immunohistochemistry analysis of human skin tissue. Tissue was fixed overnight in PBS-buffered 10% paraformaldehyde at 4°C, embedded in optimal cutting temperature compound, and frozen at −80°C. Cryosections were treated for antigen retrieval and blocking, then stained by incubation overnight at 4°C with primary antibodies in blocking solution: monoclonal mouse anti-human CCR7 (1:50) and ab10389 - polyclonal goat anti-human macrophage inflammatory protein 3 beta (1:50). Slides were incubated with fluorescent secondary goat anti-mouse Alexa488 and rabbit anti-goat Alexa647, in blocking solution (1:600) for 1 hour at room temperature. Nuclei were stained with Hoechst 33258.Four weeks of anti-TNF therapy induce lymphoid aggregate dissolution and reduction of CCR7 and CCL19 in the subpapillary dermis. Punch skin biopsy specimens were taken from the same lesional area at baseline and after 4 weeks of anti-TNF treatment (etanercept) from individual responder patients. Magnification: X40. Whi