Splenic T cells from mice undergoing GVHD are able to regenerate complex T-cell repertoires in secondary hosts. Lethally irradiated (1100 cGy) AKR mice were transplanted with Rag-1 BM (5 × 106) and a spleen cell-adjusted dose of 5 × 105 T cells obtained 19 to 29 days after BMT from B6→AKR chimeras as in Figure 1. Spleen cells from individual chimeras were each then transplanted into 2 to 3 secondary AKR hosts. AKR animals that received a secondary transplant were killed 90 days after BMT, and spleen cells were analyzed by CDR3 spectratyping. (A) Examples of skewed spectratypes of BV2, BV14, and BV16 families derived from day 19 splenocytes obtained from 3 primary GVHD (B6→AKR) mice (labeled A-C) that served as donor cells for secondary transfers (input) are shown. For comparison, unskewed spectratypes from normal B6 spleen cells are shown in the lane labeled B6. (B) Spectratypes of splenocytes from secondary AKR recipients recovered 90 days after adoptive transfer. Selected BV families from 2 secondary recipient mice (lanes 1-2) receiving GVHD T cells from either mouse A (lane A) or B (lane B) are depicted. Arrows denote skewed bands that were also present in the input splenocyte population except for BV2 where a new skew develops. The particular BV gene being analyzed is identified to the left of each set of spectratypes.
