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Blood, Vol. 110, Issue 6, 1788-1796, September 15, 2007
The immune response to lentiviral-delivered transgene is modulated in vivo by transgene-expressing antigen-presenting cells but not by CD4+CD25+ regulatory T cells
Blood Annoni et al.
110: 1788
Supplemental materials for Annoni et al, Vol 110, Issue 6, 1788-1796
Files in this Data Supplement:
- Figure S1. OVA proliferative response in mice immunized with OVA and co-injected with nTregs (PDF, 11.8 KB) -
C57BL/6 mice were immunized in the hind footpads with CFA-OVA323-339 or CFA-PBS as control. One group of mice received CFA-OVA323-339 and 2 × 106 nTregs the day before immunization. CD4+ T cells were isolated from draining lymph nodes 2 weeks later and cultured with irradiated T-cell–depleted splenocytes plus OVA323-339. OVA-specific proliferative responses were assessed by 3H incorporation 3 days after stimulation. Data are expressed as average cpm (± SD). One representative experiment out of 2 is presented (3 animals per group per experiment).
- Figure S2. CD4+ CD25+ Foxp3+ Tregs in mice co-injected with LV-CMV-GFP and GFP-tg APC (PDF, 51 KB) -
C57BL/6 mice were injected with 15 × 106 GFP-tg APC the day before LV-CMV-GFP injection. (A) Two and 4 weeks after vector administration splenocytes were isolated from mice injected with vehicle (white bars), LV-CMV-GFP (gray bars), or co-injected with LV-CMV-GFP and GFP-tg APC (black bars) and the percentages of Tregs, defined by expression of CD4 CD25 Foxp3, were assessed by FACS. Data are expressed as average percentages (± SD). (B) A representative dot plot analysis for each experimental group at the indicated time point is shown. Numbers indicate percentage of CD4+ CD25+ Foxp3+ cells in each group. Two experiments were performed to allow analysis at different time points and are both shown (the two animals in the control group were pooled from the two different experiments; 5 animals in the other groups per each time point are shown).
- Figure S3. Persistence of GFP-tg APC in secondary lymphoid organs 7 days after transfer (PDF, 63 KB) -
15 × 106 GFP-tg APC were adoptively transferred in C57BL/6 mice (as shown in Fig. 4) and euthanized at day 7. The presence of the transferred GFP-tg APC was evaluated by FACS by gating on dendritic cells (CD11c+) and B cells (B220+) co-expressing GFP. The percentages of GFP+ dendritic cells (upper plots) and GFP+ B cells (lower plots) are reported.
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