Monitoring of residual disease and guided donor leucocyte infusion after allogeneic bone marrow transplantation by chimaerism analysis with short tandem repeats

Abstract

Summary. In this study, we analysed the chimaeric status of peripheral blood leucocytes (PBLs) in recipients of allogeneic bone marrow transplantation (BMT) with the use of short tandem repeat (STR) microsatellite markers for monitoring the efficacy of BMT and donor leucocyte infusions (DLIs). A set of four STR markers was used with a highly discrimative capacity between individuals. STRs were detected by polymerase chain reaction (PCR) and were analysed by gene scanning (STR-GS). Between June 1990 and December 1998, 52 patients treated with BMT for chronic myeloid leukaemia (CML) were analysed. Seventeen patients relapsed after BMT and two patients never achieved remission after BMT. Fourteen of the 17 patients achieved a complete donor chimaerism after BMT, as detected by the presence of only donor STR-GS fragments, and in three cases a weak recipient STR-GS signal remained persistently detectable after BMT. A reappearance or increase of recipient STR-GS signals was indicative of relapse, which was mostly detected by STR-GS several months before relapse was diagnosed clinically. Nineteen patients were treated with DLI for reappearance of CML after BMT which resulted in complete remission in 17 patients, concordant with the disappearance of recipient STR-GS signals. More importantly, DLI treatment could be guided based upon the STR-GS data, which prevented unnecessary extra DLI courses that could cause toxicity. This study indicates that STR-GS is an effective and reliable method for monitoring BMT recipients.

Serial quantification of lymphoid and myeloid mixed chimerism using multiplex PCR amplification of short tandem repeat-markers predicts graft rejection and relapse, respectively, after allogeneic transplantation of CD34+ selected cells from peripheral blood

“…This might provide important information as to when early therapeutic interventions, such as donor lymphocyte infusions (DLI), should be made. 36,37 A sensitive and automatic method of quantifying the degree of MC after allo-SCT, based on PCR assays of polymorphic short tandem repeats (STR), is available. 33 Graft rejection seems to be caused by host T cells surviving the conditioning regimen.…”

How and when should we monitor chimerism after allogeneic stem cell transplantation?

“…This real-time PCR has a less quantitative accuracy with a variation coefficient of only 30-50% 54,55 compared to 5% of the STR systems. 44,46,[57][58][59] This does not hamper the analysis when only very low levels of mixed chimerism are needed to be quantified. However, in the range above 5% autologous cells, a variation coefficient of approximately 50% does not yet permit accurate quantitative investigations of the dynamic of mixed chimerism post transplant.…”

Chimerism analysis within 6 months of allogeneic stem cell transplantation predicts relapse in acute myeloid leukemia

“…Briefly, DNA was isolated from T-and non-T-cell fractions of peripheral blood samples using the salting out method. 11 STR chimerism analysis was performed as described previously 12 by using the STR markers SE33, HUMTHO, HUMVWFA, FGA, D3S1358, D19S253 and D11S554. The two most informative markers (discriminatory between donor and recipient) of these seven STR markers were used for chimerism screening after SCT using ABI Prism 377 DNA sequencer (Applied Biosystems, Foster City, CA, USA).…”