Activation of HA-specific CD4+ T cells under different immunization conditionsa
| Mice . | Immunization route . | . | Analyses at Day 4 . | . | . | . | . | . | . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | . | . | Draining LNb . | . | . | . | . | . | Nondraining LNc . | |||||||
| . | s.c. . | i.v. . | Cell number (million)d . | CD4+Thy-1.1+ (%) . | Divided cells (%)e . | CD25 (%)f . | IFN-γ (%)g . | IL-2 (%)g . | Cell number (million)d . | |||||||
| BALB/c (n = 4) | 1.3 ± 0.3 | 0.08 ± 0.03 | 6 ± 1 | 1 ± 2 | Not detected | Not detected | 2.9 ± 0.7 | |||||||||
| ins-HA (n = 13) | 2.3 ± 1.1 | 0.15 ± 0.04 | 12 ± 4 | 4 ± 1 | Not detected | Not detected | 2.9 ± 1.8 | |||||||||
| BALB/c (n = 14) | DC2 | 15.3 ± 6.3 | 2.05 ± 0.34 | 96 ± 1 | 9 ± 3 | 13.1 ± 2.9 | 7 ± 2 | 2.8 ± 0.6 | ||||||||
| BALB/c (n = 22) | DC20 | 12.1 ± 4.4 | 2.75 ± 1.1 | 97 ± 2 | 22 ± 11 | 10.3 ± 3.5 | 6 ± 3 | 2.7 ± 0.7 | ||||||||
| BALB/c (n = 13) | DC20 | CD40/LPS | 11.9 ± 4.8 | 1.90 ± 0.75 | 94 ± 3 | 57 ± 21 | 24.5 ± 6.7 | 16 ± 5 | 10 ± 3 | |||||||
| Mice . | Immunization route . | . | Analyses at Day 4 . | . | . | . | . | . | . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | . | . | Draining LNb . | . | . | . | . | . | Nondraining LNc . | |||||||
| . | s.c. . | i.v. . | Cell number (million)d . | CD4+Thy-1.1+ (%) . | Divided cells (%)e . | CD25 (%)f . | IFN-γ (%)g . | IL-2 (%)g . | Cell number (million)d . | |||||||
| BALB/c (n = 4) | 1.3 ± 0.3 | 0.08 ± 0.03 | 6 ± 1 | 1 ± 2 | Not detected | Not detected | 2.9 ± 0.7 | |||||||||
| ins-HA (n = 13) | 2.3 ± 1.1 | 0.15 ± 0.04 | 12 ± 4 | 4 ± 1 | Not detected | Not detected | 2.9 ± 1.8 | |||||||||
| BALB/c (n = 14) | DC2 | 15.3 ± 6.3 | 2.05 ± 0.34 | 96 ± 1 | 9 ± 3 | 13.1 ± 2.9 | 7 ± 2 | 2.8 ± 0.6 | ||||||||
| BALB/c (n = 22) | DC20 | 12.1 ± 4.4 | 2.75 ± 1.1 | 97 ± 2 | 22 ± 11 | 10.3 ± 3.5 | 6 ± 3 | 2.7 ± 0.7 | ||||||||
| BALB/c (n = 13) | DC20 | CD40/LPS | 11.9 ± 4.8 | 1.90 ± 0.75 | 94 ± 3 | 57 ± 21 | 24.5 ± 6.7 | 16 ± 5 | 10 ± 3 | |||||||
CFSE-labeled CD25− T cells from Thy-1.1 TCR-HA-transgenic mice were adoptively transferred in ins-HA-transgenic mice (ins-HA condition) or in BALB/c mice. The day after, BALB/c mice were immunized s.c. with DC pulsed in vitro with either 2 μg/ml HA peptide (DC2 condition), or 20 μg/ml HA peptide (DC20 condition), or 20 μg/ml HA peptide associated with i.v. injection of anti-CD40 mAb and LPS (DC20/CD40/LPS condition). Four days after activation, CD4+Thy-1.1+ donor cells were analyzed in draining LN by flow cytometry for division, expression of CD25 and intracellular IFN-γ and IL-2. Mean values ± SD are shown.
Analyses in pancreatic LN (ins-HA mice) or popliteal LN (BALB/c mice).
Mix of brachial and axillary LN.
Absolute cell numbers of viable cells per LN excluding trypan blue.
Percentages of divided cells among CD4+Thy-1.1+ were determined by loss of CFSE fluorescence.
Percentages among CD4+Thy-1.1+ cells.
Percentages among divided CD4+Thy-1.1+ cells.