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Blood, Vol. 109, Issue 12, 5318-5326, June 15, 2007
TLR3 ligand stimulates fully functional memory CD8+ T cells in the absence of CD4+ T-cell help
Blood Hervas-Stubbs et al.
109: 5318
Supplemental materials for: Hervas-Stubbs et al, Vol 109, Issue 12, 5318-5326
Files in this Data Supplement:
- Figure S1. Effect of the dose level of TLR-Ls on its ability to prime CTLs in the absence of CD4+ T cell help (PDF, 129 KB) -
C57BL/6 mice received a single intravenous. injection of BOVAp either alone or with different doses of TLR-Ls. Seven days after injection, the CTL response was quantified either by ELISPOT (A) or by in vivo killing assay (B). Dots represent individual mice. Data are representative of two independent experiments.
- Figure S2. Poly(I:C) but not TLR2/6-L or TLR4-L derived signals fulfill the requirements to prime adoptively transferred OT-1 cells (PDF, 549 KB) -
Ly5.1+ recipient mice were injected with CFSE-labeled (10 µM) OT-1 cells (1 × 106) isolated from the lymph nodes of OT-1 Rag1ko transgenic mice (95% of purity). Eighteen to twenty four hours later, Ly5.1+ recipient mice received a single intravenous. injection of PBS or BOVAp, either alone or in combination with 25 µg of poly(I:C), zymosan or LPS. Mice were sacrificed 40 or 72 hours after immunization, and their splenocytes were labeled for CD45.1 and CD8 and either CD44, CD25, CD62L or IFN- molecules. Transferred OT-1 cells were identified as CD8+ CD45.1− cells. (A) Fold expansion of OT-1 cells 40 and 72 hours after immunization of recipient mice. Absolute OT-1 cell numbers were calculated based on the percentage of OT-1 cells and the total cell counts. Fold expansion is expressed as the ratio between the absolute OT-1 cell numbers in primed mice and mice injected with PBS within each experiment. (B) Expression of effector markers in OT-1 transferred cells 40 hours (CD25, CD44 and CD62L) or 70 hours (IFN-) after immunization. For detection of intracellular IFN-, recipient mice were injected with 100 nmoles of OVA257-264 peptide 90 minutes before sacrifice. Brefeldin A was present during splenocyte isolation and staining for surface markers. Cells were then processed with the Fix & Perm kit (BD Biosciences) according to the manufacturer’s instructions and stained with anti-IFN- mAb. Results are representative of two independent experiments.
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