“…The samples were fixed for 6 h in Histochoice (Amresco, Solon, Ohio, USA), processed by an automated formalin/ethanol-based system overnight, and wax-embedded. Sections of 5 um were cut and were stained for the cytokines IL-12, IL-6 and IL-10, using goat anti-mouse IL-12, IL-6 and IL-10 primary antibodies (R&D Systems, Minneapolis, Minn., USA) followed by the secondary antibody (horse HRP-conjugated anti-goat IgG(H+L), Vector Laboratories, Burlingame, Calif., USA), stained with the chromogen 3,3′-diaminobenzidine (DAB; Kirkegaard and Perry Laboratories, Gaithersburg, Md., USA), then counter-stained briefly with hematoxylin, as previously described in detail [29,30]. The sections were examined for positive brown staining under light microscopy, and images were captured digitally using a Sony HyperHAD color video camera (Sony, Melbourne, Australia) connected to a Leica Q500MC computer (Leica Microsystems, Sydney, Australia).…”