Rockefeller University Press

Table II.

Intracellular Cytokine Profiles of Sorted ICOS⁺ and ICOS⁻CD4⁺ T Cells

pLN CD4⁺ T cells ex vivo, freshly isolated	TNF-α	IL-3	IL-6	IFN-γ	IL-2	GM-CSF	IL-13	IL-4	IL-5	IL-10
ICOS-depleted
Frequency of cells secreting a given cytokine^{^a}	79.5%	1.1%	0.12%	5.8%	4.2%	0.7%	0.7%	0.6%	0.14%	0.4%
ICOS-enriched
Frequency of cells secreting a given cytokine^{^a}	45.1%	1.4%	0.22%	13%	10%	1.8%	8.6%	8%	1.9%	11.3%
Factor of increase^{^b}	0.6×	1.3×	1.8×	2.2×	2.4×	2.6×	12.3×	13.3×	13.6×	28.3×
pLN CD4⁺ T cells ex vivo,  cultured for 6 d in vitro^{^c}
ICOS-depleted
Frequency of cells secreting a given cytokine^{^a}	84.1%	0.3%	n.d.	12.5%	7.2%	n.d.	0.21%	0.5%	0.01%	0.2%
ICOS-enriched
Frequency of cells secreting a given cytokine^{^a}	56.2%	0.5%	n.d.	18.4%	12.5%	n.d.	6.7%	9.6%	0.56%	14.9%
Factor of increase^{^b}	0.7×	1.7×	n.d.	1.5×	1.7×	n.d.	31.9×	19.2×	56×	74.5×

pLN CD4⁺ T cells ex vivo, freshly isolated	TNF-α	IL-3	IL-6	IFN-γ	IL-2	GM-CSF	IL-13	IL-4	IL-5	IL-10
ICOS-depleted
Frequency of cells secreting a given cytokine^{^a}	79.5%	1.1%	0.12%	5.8%	4.2%	0.7%	0.7%	0.6%	0.14%	0.4%
ICOS-enriched
Frequency of cells secreting a given cytokine^{^a}	45.1%	1.4%	0.22%	13%	10%	1.8%	8.6%	8%	1.9%	11.3%
Factor of increase^{^b}	0.6×	1.3×	1.8×	2.2×	2.4×	2.6×	12.3×	13.3×	13.6×	28.3×
pLN CD4⁺ T cells ex vivo,  cultured for 6 d in vitro^{^c}
ICOS-depleted
Frequency of cells secreting a given cytokine^{^a}	84.1%	0.3%	n.d.	12.5%	7.2%	n.d.	0.21%	0.5%	0.01%	0.2%
ICOS-enriched
Frequency of cells secreting a given cytokine^{^a}	56.2%	0.5%	n.d.	18.4%	12.5%	n.d.	6.7%	9.6%	0.56%	14.9%
Factor of increase^{^b}	0.7×	1.7×	n.d.	1.5×	1.7×	n.d.	31.9×	19.2×	56×	74.5×

pLN cells were activated with PMA and ionomycin for 5 h in the presence of Brefeldin A for the last 3 h and analyzed for intracellular cytokines by flow cytometry.

Obtained when comparing ICOS-depleted to ICOS-enriched cell populations.

pLN cells were magnetically sorted into ICOS-depleted and ICOS-enriched cell fractions, stimulated with PMA and ionomycin for 36 h, transferred into fresh medium, and further cultured for 6 d. At the end of culture, CD4⁺ T cells were restimulated and analyzed for intracellular cytokines as above.

n.d., not determined.

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