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Flow Cytometric Measurement of Blood Cells with BCR-ABL1 Fusion Protein in Chronic Myeloid Leukemia.
Dept. of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
Dept. of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
Dept. of Medical Science and Division of Hematology, University Hospital, Uppsala, Sweden.
Department of Anthropology, University of Vienna, Vienna, Austria.
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2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, no 1, 623Article in journal (Refereed) Published
Abstract [en]

Chronic myeloid leukemia (CML) is characterized in the majority of cases by a t(9;22)(q34;q11) translocation, also called the Philadelphia chromosome, giving rise to the BCR-ABL1 fusion protein. Current treatment with tyrosine kinase inhibitors is directed against the constitutively active ABL1 domain of the fusion protein, and minimal residual disease (MRD) after therapy is monitored by real-time quantitative PCR (RQ-PCR) of the fusion transcript. Here, we describe a novel approach to detect and enumerate cells positive for the BCR-ABL1 fusion protein by combining the in situ proximity ligation assay with flow cytometry as readout (PLA-flow). By targeting of the BCR and ABL1 parts of the fusion protein with one antibody each, and creating strong fluorescent signals through rolling circle amplification, PLA-flow allowed sensitive detection of cells positive for the BCR-ABL1 fusion at frequencies as low as one in 10,000. Importantly, the flow cytometric results correlated strongly to those of RQ-PCR, both in diagnostic testing and for MRD measurements over time. In summary, we believe this flow cytometry-based method can serve as an attractive approach for routine measurement of cells harboring BCR-ABL1 fusions, also allowing simultaneously assessment of other cell surface markers as well as sensitive longitudinal follow-up.

Place, publisher, year, edition, pages
Springer Nature , 2017. Vol. 7, no 1, 623
National Category
Medical Genetics Cell and Molecular Biology
Research subject
Biomedicine
Identifiers
URN: urn:nbn:se:oru:diva-57339DOI: 10.1038/s41598-017-00755-yISI: 000398162400034PubMedID: 28377570ScopusID: 2-s2.0-85016991093OAI: oai:DiVA.org:oru-57339DiVA: diva2:1102657
Note

Funding agencies:

DIATOOLS 259796

GastricGlycoExplorer 316929

PRIMES 278568

European Research Council under the European Union's Seventh Framework Programme (FP/2007–2013) ProteinSeq (294409)

IngaBritt och Arne Lundbergs Forskningsstiftelse

Swedish Cancer Society Swedish Research Council

Available from: 2017-05-30 Created: 2017-05-30 Last updated: 2017-05-30Bibliographically approved

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