Reconstitution of T cell receptor repertoire after human allogeneic bone marrow transplantation

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Reconstitution of T cell receptor (TCR) repertoire after human allogeneic marrow transplantation was investigated. Informed consent was obtained before drawing blood samples from the patients and their marrow donors. TCR Vbeta and Valpha repertoire was analyzed by using adaptor ligation PCR and microplate hybridization assay techniques. There was no significant difference in TCR repertoire between donor-recipient. After8-10 weeks of transplant, a striking uncrease of the T cells carrying certain Vbeta and Valpha subfamilies was observed in all patients examined. Distorted TCR repertoire appears to retum to the normal pattem after 1 year of transplant. With the similar kinetics to the TCR repertoire changes, there was an increase of CD8+CD28-HLA-DR+T lymphocytes after transplant. Vbeta and Valpha chains with increased frequency were preferentially used in this lymphocyte subset. Taken together, the proliferation of CD8+CD28-HLA-DR+T lymphocytes results in the distortion of TCR repertoire after transplant.To investigate the TCR diversity of blood lymphocytes in marrow recipients, CDR3 size spectratyping analsis was also performed in several patients. The lack of multiple Vbeta subfamilies and the disturbed recovery of CDR3 complexity were observed in the patient with cytomegalovirus pneumonia, suggesting that this method might be useful for monitoring immune reconstitution after marrow transplantation.