Table I.

Th17 cells differentiated with IL-6 + TGF-β are not encephalitogenic after primary stimulation

 Cytokinesa    
T cell differentiation IFN-γ IL-17 Incidence of EAE Mean day of onset Mean clinical score on day of onset 
Primary stimulation      
 Th17 (IL-6 + TGF-β) − ++ 0/28 (0%) NA NA 
 Th17 (IL-6 + TGF-β + IL-23) ++ 1/12 (8.3%) NA NA 
 Th17 (IL-6 + TGF-β) + PTXb − ++ 0/7 (0%) NA NA 
 Th17 (IL-6 + TGF-β) + Th1c − ++ 0/4 (0%) NA NA 
 Th17 (IL-6 + αIL-12/IFN-γ/IL-4) − 17/17 (100%) 7.6 2.2 
 Th17 (IL-6 + αIL-12/IFN-γ/IL-4/TGF-β) − 4/5 (80%) 6.3 1.5 
Secondary stimulationd      
 Th17 (IL-6 + TGF-β) +++ +++ 10/13 (76.9%) 6.2 2.8e 
 Th17 (IL-6 + TGF-β + IL-23) +++ +++ 6/7 (85.7%) 2.3 
 Th17 (IL-6 + TGF-β → IL-23)f +++ +++ 3/5 (60%) 11.3 1.3 
 Th17 (IL-6 + TGF-β → IL-1β)f +++ +++ 4/5 (80%) 11 2.3 
 Cytokinesa    
T cell differentiation IFN-γ IL-17 Incidence of EAE Mean day of onset Mean clinical score on day of onset 
Primary stimulation      
 Th17 (IL-6 + TGF-β) − ++ 0/28 (0%) NA NA 
 Th17 (IL-6 + TGF-β + IL-23) ++ 1/12 (8.3%) NA NA 
 Th17 (IL-6 + TGF-β) + PTXb − ++ 0/7 (0%) NA NA 
 Th17 (IL-6 + TGF-β) + Th1c − ++ 0/4 (0%) NA NA 
 Th17 (IL-6 + αIL-12/IFN-γ/IL-4) − 17/17 (100%) 7.6 2.2 
 Th17 (IL-6 + αIL-12/IFN-γ/IL-4/TGF-β) − 4/5 (80%) 6.3 1.5 
Secondary stimulationd      
 Th17 (IL-6 + TGF-β) +++ +++ 10/13 (76.9%) 6.2 2.8e 
 Th17 (IL-6 + TGF-β + IL-23) +++ +++ 6/7 (85.7%) 2.3 
 Th17 (IL-6 + TGF-β → IL-23)f +++ +++ 3/5 (60%) 11.3 1.3 
 Th17 (IL-6 + TGF-β → IL-1β)f +++ +++ 4/5 (80%) 11 2.3 

NA, not applicable.

a

Cytokine production was measured by ELISA at 72 h in culture during differentiation or following restimulation with MBP Ac1-11 only for secondary stimulation. +, <1 ng/ml; ++, 1–4 ng/ml; +++, >4 ng/ml.

b

Mice were administered 200 ng pertussis toxin (PTX) on days 0 and 2 after transfer of Th17 (IL-6 + TGF-β) cells into mice.

c

Mice were injected with 5 × 106 Th17 (IL-6 + TGF-β) cells plus 106 Th1 (IL-12) cells.

d

Th17 cells were rested for 4 d after differentiation and restimulated with MBP Ac1-11 only before transfer into naive B10.PL mice.

e

The rapid onset and severity of the disease required that the mice be euthanized within 48 h of disease onset, and therefore, only the mean clinical score at the time of disease onset could be determined.

f

Th17 cells (IL-6 + TGF-β) were rested for 4 d after differentiation and restimulated with MBP Ac1-11 + IL-23 or IL-1β before transfer into naive B10.PL mice.

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