. Author manuscript; available in PMC: 2015 May 21.

Published in final edited form as: Comput Methods Programs Biomed. 2008 Jul 21;92(1):54–65. doi: 10.1016/j.cmpb.2008.06.002

Table III.

Comparison of cytokine-secreting cells measured by intracellular staining (ICS) and Fluorospot.

		IL2+/IFNγ−	IL2+/IFNγ+	IL2−/IFNγ+
Fluorospot	No Ag	13.3 ± 8.1	4.0 ± 2.0	4.0 ± 2.0
	Influenza	133.3 ± 28.4	90.7 ± 14.0	114.7 ± 15.1
ICS	No Ag	3.0 ± 1.3	0.8 ± 0.6	3.7 ± 0.8
	Influenza	7.8 ± 5.0	22.2 ± 8.5	26.5 ± 11.0

Human PBMC were stimulated with influenza antigen or left unstimulated, and cytokine secretion or synthesis were measured in triplicate by Fluorospot or intracellular cytokine staining, respectively. Single and double-producing cells were measured by analysis of ICS results in FlowJo, gating on CD4+ T cells (there was no CD8 T cell response to the influenza vaccine antigen), and Fluorospot results were analyzed in Exploraspot. The table shows cytokine-secreting cell frequencies per million PBMC added to cultures. Similar results were obtained with a second subject.