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Table II.

Intracellular Cytokine Profiles of Sorted ICOS+ and ICOSCD4+ 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 cytokinea 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 cytokinea 45.1% 1.4% 0.22% 13% 10% 1.8% 8.6% 8% 1.9% 11.3% 
Factor of increaseb 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 vitroc           
ICOS-depleted           
Frequency of cells secreting a given cytokinea 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 cytokinea 56.2% 0.5% n.d. 18.4% 12.5% n.d. 6.7% 9.6% 0.56% 14.9% 
Factor of increaseb
 
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 cytokinea 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 cytokinea 45.1% 1.4% 0.22% 13% 10% 1.8% 8.6% 8% 1.9% 11.3% 
Factor of increaseb 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 vitroc           
ICOS-depleted           
Frequency of cells secreting a given cytokinea 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 cytokinea 56.2% 0.5% n.d. 18.4% 12.5% n.d. 6.7% 9.6% 0.56% 14.9% 
Factor of increaseb
 
0.7×
 
1.7×
 
n.d.
 
1.5×
 
1.7×
 
n.d.
 
31.9×
 
19.2×
 
56×
 
74.5×
 
a

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.

b

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

c

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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