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.