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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.

A PCR method was developed to analyse each of 29 families of the T cell receptor V alpha gene and 20 families of the V beta gene at the mRNA level in heterogenous cell populations. All V alpha and V beta families were detectable in blood mononuclear cells from four of six healthy donors. In two donors only V alpha 22 was missing, and all other V alpha and V beta families were detected. V beta family expression was observed in T-leukaemic cell lines Jurkat, HSB, Molt-3 and Molt-4. In contrast, V alpha family expression was not detectable in any cell line except Jurkat cells. In T-cell malignancies (non-Hodgkin's lymphoma and mycosis fungoides), one or two V alpha and V beta families were detectable. Four of 10 cases investigated showed two V alpha transcripts and one V beta transcript. This fits with concepts in literature that allelic exclusion for the genes encoding alpha chains is not strictly required in the DNA rearrangement, or that this exclusion is a post-translational event. Using a limited series of antibodies to V beta gene family products, blood mononuclear cells from healthy donors were analysed by flow cytometry in a follow-up study. Two of four donors were rather stable in proportions of T cells expressing distinct V beta families, and two other donors showed variation in one or more families. When analysed on frozen tissue sections of normal lymph node and tonsil, there was no preferential location of lymphocytes expressing a distinct V beta gene family in different compartments (interfollicular area, follicle, or tonsillar epithelium).