Multiparametric flow cytometry is an alternative approach to the polymerase chain reaction method for evaluating minimal residual disease in treatment protocols for primary acute lymphoblastic leukemia. Given considerable differences between primary and relapsed acute lymphoblastic leukemia treatment regimens, flow cytometric assessment of minimal residual disease in relapsed leukemia requires an independent comprehensive investigation. In the present study we addressed evaluation of minimal residual disease by flow cytometry in the clinical trial for childhood relapsed acute lymphoblastic leukemia using eight-color flow cytometry. The major challenge of the study was to reliably identify low amounts of residual leukemic cells against the complex background of regeneration, characteristic of follow-up samples during relapse treatment. In a prospective study of 263 follow-up bone marrow samples from 122 patients with B-cell precursor acute lymphoblastic leukemia, we tested various B-cell markers, adapted the antibody panel to the treatment protocol, and evaluated its performance by a blinded parallel comparison with the polymerase chain reaction data. The resulting eight-color single-tube panel showed a consistently high overall concordance (P<0.001) and, under optimal conditions, sensitivity similar to that of the reference polymerase chain reaction method. Overall, evaluation of minimal residual disease by flow cytometry can be successfully integrated into the clinical management of relapsed childhood acute lymphoblastic leukemia either as complementary to the polymerase chain reaction or as an independent risk stratification tool. ALL-REZ BFM 2002 clinical trial information: NCT00114348.
- MeSH
- antitumorózní látky terapeutické užití MeSH
- B-lymfocyty účinky léků imunologie patologie MeSH
- CD antigeny genetika imunologie MeSH
- DNA-nukleotidylexotransferasa genetika imunologie MeSH
- exprese genu MeSH
- imunofenotypizace MeSH
- kojenec MeSH
- kostní dřeň účinky léků imunologie patologie MeSH
- lidé MeSH
- nádorové biomarkery genetika imunologie MeSH
- polymerázová řetězová reakce MeSH
- pre-B-buněčná leukemie diagnóza farmakoterapie genetika patologie MeSH
- předškolní dítě MeSH
- protoonkogenní proteiny c-bcl-2 genetika imunologie MeSH
- průtoková cytometrie metody MeSH
- recidiva MeSH
- reziduální nádor MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- práce podpořená grantem MeSH
We have used selected rabbit anti-human polyclonal antibodies as an example of useful and easily available tools for studies on immune system structure and development in important veterinary species, many of which also represent animal models in biomedicine. The cocktail of anti-human Igkappa-FITC/anti-Iglambda-RPE F(ab')(2) fragments was used for two-colour and, in combination with the cross-reactive anti-CD79alpha monoclonal antibody HM-57, for three-colour flow cytometry of canine, feline, bovine and porcine peripheral B-cells. A possible application of such immunoreagents in studies on primary B-cell differentiation has been suggested in pigs; the same approach can be used in other species of interest. Rabbit anti-human lactoferrin-FITC F(ab')(2) fragment was used for visualizing neutrophils in dogs, pigs and cattle and an application for two-colour immunophenotyping of canine granulocyte subsets has been designed. Affinity isolated rabbit anti-human CD3 and anti-human TdT have been shown to represent a ready-to-use tool for in situ studies on primary T-lymphopoiesis in pigs with possible extensions both to the B-lineage development in pigs and other animal models. Altogether, our study show that carefully selected polyclonal antibodies available on the market may possess broad cross-reactivity with important applications in veterinary research.
- MeSH
- antisérum imunologie MeSH
- CD antigeny analýza imunologie MeSH
- DNA-nukleotidylexotransferasa analýza imunologie MeSH
- financování organizované MeSH
- imunitní systém fyziologie MeSH
- imunohistochemie MeSH
- kočky MeSH
- králíci MeSH
- lehké řetězce imunoglobulinů analýza imunologie MeSH
- lidé MeSH
- prasata MeSH
- průtoková cytometrie MeSH
- psi MeSH
- skot MeSH
- zkřížené reakce MeSH
- zvířata MeSH
- Check Tag
- kočky MeSH
- králíci MeSH
- lidé MeSH
- psi MeSH
- skot MeSH
- zvířata MeSH
