Figure 5

Adoptive transfer of CD4⁺ T cells from line 18 mice in BALB/c recipient. CD4⁺ T cells were isolated from spleen and lymph nodes of DO11.hCARΔcyt line 13 transgenic mice, transduced for 30' with Ad5.UbP.GFP at a MOI of 10 and incubated for 24 h at 37°C/5%CO₂. 5 × 10⁶ transduced cells were injected in the tail vein of BALB/c mice. The mice were sacrificed after 48 h and the inguinal, axillary, submandibular and mesenteric lymph nodes, the spleen and the Peyer's Patches were isolated. A. Flow cytometric analysis of GFP expression of transduced CD4⁺ cells before and after adoptive transfer. Recovered, pooled lymph node cells from BALB/c recipients were stained with anti-CD4 and anti-DO11.10 TCR clonotypic mAb, KJ-126, and analyzed on a FACScan. The panels depict two-color dot plots of the transduced CD4⁺ cells before transfer (top panel) and the pooled lymph node cells from a representative BALB/c recipient (lower panel) and histograms representing the GFP expression for each of the CD4⁺KJ126⁺ population (square gate). The percentage of positive cells is indicated. B. Immunohistochemical analysis of a peripheral lymph node of Ad5.UbP.GFP-transduced CD4 T cell adoptive transfer recipient. An inguinal lymph node was processed and stained with anti-B22O (red) as described in Material and Methods. The red fluorescent areas define primary B cell follicles. GFP positive cells (green) are indicated (arrows) in parafollicular, T cell areas of a representative lymph node (magnification: top, 10×; bottom, 40×).