Expression of mRNA and protein for MIP-1α at days 6 and 13 after syngeneic and allogeneic T-cell transfer.

RNA was isolated from the individual organs, and RPA was performed as described in the text. Expression of MIP-1α was analyzed by densitometry using the NIH Image Software Program, then normalized to the expression of GAPDH for each condition. The mean expression with SE is given. Background activity was subtracted using an internal control for each lane. We evaluated 4 mice for each condition. Each experiment was repeated 3 times, and the data were pooled from the 3 experiments. Control animals received irradiation and syngeneic splenocytes. (A) Data are shown for expression at day 6 from the GI tract, liver, lung, and spleen. (B) The same analysis was performed as described above except that the mice were killed 13 days after splenocyte transfer. We evaluated 3 mice per condition, and each experiment was repeated twice. The results are given as the mean expression and SE and are pooled from 2 separate experiments. (C) Expression of MIP-1α at day 13 in GVHD target organs was evaluated as above and compared to the expression in the kidney, heart, and pancreas. (D) ELISA was performed on colonic, hepatic, pancreatic, and splenic tissue from bm1 and bm12 recipients on day 6 following the transfer. The limit of detection of MIP-1α for the assay is 3.75 pg/mL. We evaluated 3 mice per condition. *P < .001; **P = .02 for comparison between class I and class II only.