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Blood, Vol. 113, Issue 18, 4449-4457, April 30, 2009
Depletion of CD4 T cells enhances immunotherapy for neuroblastoma after syngeneic HSCT but compromises development of antitumor immune memory
Blood Jing et al.
113: 4449
Supplemental materials for: Jing et al
Files in this Data Supplement:
- Figure S1. Altered CD4/CD8 T-cell ratios in the spleens and lymph nodes of mice early after HSCT (JPG, 95.6 KB)
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(A) T cells were isolated from the spleens of tumor-vaccinated donors (sensitized T cells) by negative selection using immunomagnetic sorting and analyzed by flow cytometry for CD8 and CD4 expression. (B) Administration of anti-CD4 mAb effectively depleted CD4+ T cells from the lymphoid tissues of treated mice. Analysis of CD8 and CD4 T cells in the lymph nodes (LN) and spleens from tumor-bearing mice given HSCT and 6 × 106 tumor-sensitized T cells (Sen T), ± AGN2a-4P vaccine (Vac), and ± anti-CD4 mAb treatment. The anti-CD4 mAb was administered 1 day before HSCT, and the AGN2a-4P vaccine was given 2 days after HSCT. Cells were harvested for flow cytometric analysis 4 days after HSCT. (C) Most CD4 T cells in the spleens of HSCT recipient mice early after transplantation were endogenous CD4 T cells that survived the total body irradiation. Splenocytes collected from mice given HSCT and 6 × 106 CFSE-labeled tumor-sensitized T cells (Sen T) were analyzed by flow cytometry 4 days after HSCT. CFSE labeling was used to distinguish the endogenous CD4 T cells from the CD4 T cells transferred with the graft.
- Figure S2. Elevated percentages of CD4+CD25+Foxp3+ T cells in the lymphoid tissues of mice given HSCT and tumor-sensitized T cells (JPG, 63.7 KB)
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Splenocytes were isolated from the spleens of mice 14 days after HSCT and T-cell transfer (6 × 106 tumor-sensitized T cells), and the cells were analyzed by flow cytometry for CD4, CD25, and Foxp3 expression. The 2-color histograms depict the expression of Foxp3 and/or CD25 on electronically gated CD4+ T cells.
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