Summary:We have examined the reconstitution of ␥␦ T cell repertoire diversity after human allogeneic hematopoietic cell transplantation using a polymerase chain reaction (PCR)-based complementarity-determining region (CDR) 3 size spectratyping and DNA sequencing. The CDR3 complexity in the variable region of the T cell receptor (TCR)-␦ chain was different amongst the individuals studied. Furthermore, CDR3 size distribution patterns of allogeneic hematopoietic cell transplant recipients were almost completely recovered by a few months after transplantation. In some patients, clonal predominance of the TCRDV1+ T cells became evident during the period after transplantation. In one particular donor/recipient pair, clonal predominance of TCRDV1+ T cells was already present in blood lymphocytes of the donor, and was also observed in the recipient after transplantation. Using this donor/recipient pair, we have questioned whether ␥␦ T cell regeneration occurs via the peripheral expansion of mature T cells in the graft. In the donor lymphocytes, two expanding ␥␦ T cell clones, which were demonstrated by CDR3 sequences of the TCR-␦ chain, were recognized. These two clones were identified in the T cells from the recipient post transplant, but not before transplantation. One of the two clones was still detectable 1. years after the transplant procedure. These results strongly suggest that peripheral expansion of mature T cells in the graft is the principal pathway of ␥␦ T cell regeneration after allogeneic hematopoietic cell transplantation in adults. Bone Marrow Transplantation (2000) 26, 177-185. Keywords: T cell receptor; ␦-chain; diversity; transplantation High-dose chemoradiotherapy depletes T cells in blood and marrow transplant recipients and adequate T cell regeneration is essential for successful transplantation. The repertoire of T cells requires to be sufficiently diverse in order to recognize a wide range of microorganism-derived antigens. ␣ T cells are almost exclusively thymus-dependent