Table 2.

Summary of current gating recommendations for assay harmonization

	RECOMMENDATION
Cytokine gate	Cytokine gate should not include negative cells and should include all of the cytokine-positive cells (both high- and low-positive cells).
Double-positive cells	Gate on CD4 and CD8 to allow clear identification of the double-positive cells. Include as a separate population or with either CD4 or CD8 cells but not with both.
Dim cells	Draw gates around the main populations of CD3, CD4, and CD8 that include dim events.
Lymphocyte gate	Lymphocyte gate should be large enough to include all lymphocytes. It should not include RBC or debris.
Uniform gates	Uniform gates should be used for all samples within a donor.
Biexponential scaling	Proper adjustment of biexponential scaling is important to clearly and completely visualize all populations.