Lymphoid Reconstitution After Autologous PBSC Transplantation With FACS-Sorted CD34+ Hematopoietic Progenitors

T-cell and B-cell reconstitution was studied in nine patients who received fluorescence activated cell sorter (FACS)-sorted autologous CD34+ hematopoietic progenitor cells (HPC). The mean numbers of T cells (CD3+), B cells (CD19+) and CD34+ HPC administered to each patient were .004, .002, and 1.8 x 10(6) cells/kg, respectively. After high-dose myeloablative chemotherapy (busulfan, cyclophosphamide, etoposide) CD34+) HPC were infused and lymphoid reconstitution was monitored using flow cytometry and reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of VDJ T-cell receptor (TcR) sequences. Restoration of normal numbers of peripheral blood T cells and B cells among recipients of FACS-sorted CD34+ HPC was delayed compared to recipients of non-T-cell-depleted PBSC autografts. In both patient groups, the circulating T cells were primarily CD4-, CD8+, alphabeta TcR+, and CD45RO+, CD45RA- during the first 2 months after transplant. Subsequent increases in the frequency of CD45RA+ CD45RO- T cells occurred at 2 to 3 months after transplant, suggesting maturation of CD34+ hematopoietic progenitors to "naive" T cells. Analysis of the TcR repertoire after hematopoietic reconstitution demonstrated decreased diversity of Vbeta TcR expression associated with global decreases in the absolute number of total peripheral blood T cells and most Vbeta TcR+ subsets. Three of nine recipients of FACS-sorted CD34+ HPC demonstrated significant increases in the percentage of gammadelta+ peripheral T cells and CD5+ B cells at 3 to 9 weeks after transplantation, and all patients had transient oligoclonal expansions of T cells expressing specific Vbeta TcR. Transplantation with highly purified CD34+ HPC results in reduced diversity of the peripheral T-cell repertoire during the early post-transplant period compared with patients receiving unmanipulated or MoAb-depleted transplants.