Allo-reactivity modulates the migration and persistence of allo-T-bodies. (A) Comparative in vivo bioluminescence imaging of Renca-Her2/neu-bearing mice (n = 6/group) treated with either syngeneic (Balb-N29) or allogeneic (Black-N29) T-bodies according to different protocols, as indicated. Day 1 indicates 1 day after the transfer of the second cell batch. BLI was performed as described in “Methods.” Images shown are of 3 representative mice per group. 200R/30 = Irradiation with 200 rads and 30 × 10⁶ T-bodies. 200R/100 = Irradiation with 200 rads and 100 × 10⁶ T-bodies. 400R/30 = Irradiation with 400 rads and 30 × 10⁶ T-bodies. (B) Whole body BLI signal intensities from sequential imaging depicted in panel A. Each line represents a single animal. Pairwise differences between groups were analyzed using the Mann-Whitney test. P = .005 for Black-N29 vs Balb-N29 using the 200R/100 protocol on day 1. P = .005 for Black-N29 vs Balb-N29 using the 400R/30 protocol on day 7. P = .002 for Black-N29 vs Balb-N29 using the 400R/30 protocol on day 14. (C) In vivo proliferative capacity of allogeneic and syngeneic T-bodies. CFSE-labeled T-bodies were systemically transferred to preconditioned (200R or 400R) tumor-bearing mice according to the protocols described. Splenocytes of mice were harvested 4 days later and stained with an anti-idiotypic antibody to identify donor T-bodies. CFSE staining is shown for the transferred T-bodies. (D) CD3⁺ splenocytes from mice treated with allo–T-bodies using the 400R/30 protocol analyzed for expression of CFSE vs CD62L. Progressive differentiation (loss of CD62L) was concomitant with proliferation (loss of CFSE staining), P = .0001 using the χ² test.