Cells stained with pSTAT-1 antibodies were classified as macrophages M1 and those with CD163, CD206, and CMAF staining were classified as M2.^7,11The cells were quantified in duplicate, and the results were expressed as number of stained cells per square millimeter. The quantification method was the same as that used in a previous study conducted by our group.⁴ The number of cells immunostained with specific antibodies in the superficial and middle dermis (portion of the epidermis where the pathophysiological events of urticaria occur) was enumerated at a magnification of 400× in 12 optical fields, using an optical microscope (Zeiss™, Axiophot, Germany). The density of immunostained cells was calculated with the following equation:

Cell density/mm² = (immunostained cells in 12 fields)/12/0.0625.

The variability of the duplicate count was not greater than 5%.

Blank immunohistochemistry stains were performed, where the primary antibodies were excluded, which were negative.