Anti cd19

Dual immunofluorescence was performed to colocalize CD4, CD19, IL-17A and IL-22 to AhR. Briefly, optimum cutting temperature embedded skin tissues were cut into 5-μm sections and boiled in ethylenediaminetetraacetic acid buffer (pH, 9.0) for antigen retrieval. The sections were incubated with the primary antibody overnight at 4°C. The following primary antibodies (mouse antihuman) were used: anti-CD4 (Servicebio, Wuhan, China, 1:100), anti-CD19 (Servicebio, Wuhan, China, 1:100), anti-IL-17A (Proteintech, Beijing, China, 1:200), anti-IL-22 (Bioss, Beijing, China, 1:100) and anti-AhR (Abcam, Cambridge, US). Then, the sections were incubated with corresponding fluorescence secondary antibody and counterstained using 4’,6-diamidino-2- phenylindole. The sections were observed under an inverted fluorescent microscope (Nikon, Tokyo, Japan) at original magnification ×200 under appropriate absorption/emission wavelengths (Alexa Fluor, 594 nm or 488 nm). The images were captured using a Nikon Digital Still Camera and analyzed by 2 independent investigators. Immunostained cells in the dermis were counted, and results were expressed as the number of positive cells/high power field (HPF). Interobserver variations for the counts were <10%.