Allogeneic stem cell transplantation (alloSCT) is used for treating patients with T-prolymphocytic leukemia (T-PLL). However, direct evidence of GvL activity in T-PLL is lacking. We correlated minimal residual disease (MRD) kinetics with immune interventions and T-cell receptor (TCR) repertoire diversity alterations in patients after alloSCT for T-PLL. Longitudinal quantitative MRD monitoring was performed by clone-specific real-time PCR of TCR rearrangements (n=7), and TCR repertoire diversity assessment by next-generation sequencing (NGS; n=3) Although post-transplant immunomodulation (immunosuppression tapering or donor lymphocyte infusions) resulted in significant reduction (>1 log) of MRD levels in 7 of 10 occasions, durable MRD clearance was observed in only two patients. In all three patients analyzed by TCR-NGS, MRD responses were reproducibly associated with a shift from a clonal, T-PLL-driven profile to a polyclonal signature. Novel clonotypes that could explain a clonal GvL effect did not emerge. In conclusion, TCR-based MRD quantification appears to be a suitable tool for monitoring and guiding treatment interventions in T-PLL. The MRD responses to immune modulation observed here provide first molecular evidence for GvL activity in T-PLL which, however, may be often only transient and reliant on a poly-/oligoclonal rather than a monoclonal T-cell response.

• MeSH
• buněčné klony imunologie   MeSH
• dospělí MeSH
• genová přestavba T-lymfocytů genetika   MeSH
• homologní transplantace MeSH
• imunomodulace * MeSH
• kinetika MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• reakce štěpu proti leukémii * MeSH
• receptory antigenů T-buněk analýza   genetika   MeSH
• reziduální nádor diagnóza   genetika   MeSH
• senioři MeSH
• T-buněčná prolymfocytární leukemie diagnóza   terapie   MeSH
• transplantace kmenových buněk metody   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• receptory antigenů T-buněk MeSH

In chronic lymphocytic leukemia (CLL) the level of minimal residual disease (MRD) after therapy is an independent predictor of outcome. Given the increasing number of new agents being explored for CLL therapy, using MRD as a surrogate could greatly reduce the time necessary to assess their efficacy. In this European Research Initiative on CLL (ERIC) project we have identified and validated a flow-cytometric approach to reliably quantitate CLL cells to the level of 0.0010% (10(-5)). The assay comprises a core panel of six markers (i.e. CD19, CD20, CD5, CD43, CD79b and CD81) with a component specification independent of instrument and reagents, which can be locally re-validated using normal peripheral blood. This method is directly comparable to previous ERIC-designed assays and also provides a backbone for investigation of new markers. A parallel analysis of high-throughput sequencing using the ClonoSEQ assay showed good concordance with flow cytometry results at the 0.010% (10(-4)) level, the MRD threshold defined in the 2008 International Workshop on CLL guidelines, but it also provides good linearity to a detection limit of 1 in a million (10(-6)). The combination of both technologies would permit a highly sensitive approach to MRD detection while providing a reproducible and broadly accessible method to quantify residual disease and optimize treatment in CLL.

• MeSH
• CD antigeny metabolismus   MeSH
• chronická lymfatická leukemie patologie   terapie   MeSH
• dospělí MeSH
• imunofenotypizace MeSH
• kombinovaná terapie MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• následné studie MeSH
• prognóza MeSH
• průtoková cytometrie normy   MeSH
• reziduální nádor diagnóza   genetika   metabolismus   MeSH
• staging nádorů MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• CD antigeny MeSH

BACKGROUND: The knowledge of biological characteristics of minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) remains sparse. There are no data available on what level of MRD might be 'safe' without an overt risk of relapse, or whether any such level exists at all. To address this issue in prospective studies, we have developed a quantitative molecular approach to monitor MRD in CLL, which allows the malignant clone to be traced with far higher sensitivity than possible with the techniques available currently. METHOD: To quantify MRD in CLL patients, a novel locked nucleic acid (LNA)-RNA-based quantitative real-time PCR technique was developed. Clone-specific assays were prepared for 62 CLL patients. Thirty patients were followed up molecularly for a median of 250 days (range 69-570 days). All patients were administered chemo/immunotherapy. RESULTS: In three patients, molecular negativity was achieved, as estimated by LNA-based assays. In one patient, a sustained molecular negativity was established by chemo/immunotherapy and the patient remains molecularly negative (322 days). The LNA-based assay enabled us to evaluate MRD in a reproducible manner with the sensitivity of 10(-7). CONCLUSION: LNA-RNA-based quantitative real-time PCR is an effective approach for MRD monitoring with the potential for increased sensitivity compared with standard DNA-based assays used for molecular follow-up.

• MeSH
• antigeny CD38 genetika   metabolismus   MeSH
• chronická lymfatická leukemie diagnóza   genetika   MeSH
• diagnostické techniky molekulární metody   MeSH
• hybridizace nukleových kyselin metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• polymerázová řetězová reakce metody   MeSH
• prospektivní studie MeSH
• protein-tyrosinkináza ZAP-70 genetika   metabolismus   MeSH
• průtoková cytometrie MeSH
• reprodukovatelnost výsledků MeSH
• reziduální nádor diagnóza   genetika   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• těžké řetězce imunoglobulinů genetika   metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD38 MeSH
• nádorový supresorový protein p53 MeSH
• protein-tyrosinkináza ZAP-70 MeSH
• těžké řetězce imunoglobulinů MeSH