A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study
Identifieur interne : 000724 ( Pmc/Curation ); précédent : 000723; suivant : 000725A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study
Auteurs : A C Rawstron [Royaume-Uni] ; C. Fazi [Italie] ; A. Agathangelidis [Italie] ; N. Villamor [Espagne] ; R. Letestu [France] ; J. Nomdedeu [Espagne] ; C. Palacio [Espagne] ; O. Stehlikova [République tchèque] ; K-A Kreuzer [Allemagne] ; S. Liptrot [Irlande (pays)] ; D. O'Brien [Irlande (pays)] ; R M De Tute [Royaume-Uni] ; I. Marinov [République tchèque] ; M. Hauwel [Suisse] ; M. Spacek [République tchèque] ; J. Dobber [Pays-Bas] ; A P Kater [Pays-Bas] ; P. Gambell [Australie] ; A. Soosapilla [Australie] ; G. Lozanski [États-Unis] ; G. Brachtl [Autriche] ; K. Lin [Royaume-Uni] ; J. Boysen [États-Unis] ; C. Hanson [États-Unis] ; J L Jorgensen [États-Unis] ; M. Stetler-Stevenson [États-Unis] ; C. Yuan [États-Unis] ; H E Broome [États-Unis] ; L. Rassenti [États-Unis] ; F. Craig [États-Unis] ; J. Delgado [Espagne] ; C. Moreno [Espagne] ; F. Bosch [Espagne] ; A. Egle [Autriche] ; M. Doubek [République tchèque] ; S. Pospisilova [République tchèque] ; S. Mulligan [Australie] ; D. Westerman [Australie] ; C M Sanders [États-Unis] ; R. Emerson [États-Unis] ; H S Robins [États-Unis] ; I. Kirsch [États-Unis] ; T. Shanafelt [États-Unis] ; A. Pettitt [Royaume-Uni] ; T J Kipps [États-Unis] ; W G Wierda [États-Unis] ; F. Cymbalista [France] ; M. Hallek [Allemagne] ; P. Hillmen [Royaume-Uni] ; E. Montserrat [Espagne] ; P. Ghia [Italie]Source :
- Leukemia [ 0887-6924 ] ; 2016.
Abstract
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.
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DOI: 10.1038/leu.2015.313
PubMed: 26639181
PubMed Central: 4832072
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Fazi</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Agathangelidis, A" sort="Agathangelidis, A" uniqKey="Agathangelidis A" first="A" last="Agathangelidis">A. Agathangelidis</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Villamor, N" sort="Villamor, N" uniqKey="Villamor N" first="N" last="Villamor">N. 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Dobber</name><affiliation wicri:level="1"><nlm:aff id="aff13"><institution>Department of Hematology, Academic Medical Centre</institution>, Amsterdam,<country>Netherlands</country></nlm:aff><country xml:lang="fr">Pays-Bas</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kater, A P" sort="Kater, A P" uniqKey="Kater A" first="A P" last="Kater">A P Kater</name><affiliation wicri:level="1"><nlm:aff id="aff13"><institution>Department of Hematology, Academic Medical Centre</institution>, Amsterdam,<country>Netherlands</country></nlm:aff><country xml:lang="fr">Pays-Bas</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gambell, P" sort="Gambell, P" uniqKey="Gambell P" first="P" last="Gambell">P. Gambell</name><affiliation wicri:level="1"><nlm:aff id="aff14"><institution>Peter MacCallum Cancer Centre</institution>, Melbourne,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Soosapilla, A" sort="Soosapilla, A" uniqKey="Soosapilla A" first="A" last="Soosapilla">A. Soosapilla</name><affiliation wicri:level="1"><nlm:aff id="aff15"><institution>Hematology and Flow Cytometry, Laverty Pathology</institution>, Sydney,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lozanski, G" sort="Lozanski, G" uniqKey="Lozanski G" first="G" last="Lozanski">G. Lozanski</name><affiliation wicri:level="1"><nlm:aff id="aff16"><institution>Ohio State University Medical Center</institution>, Columbus, OH,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Brachtl, G" sort="Brachtl, G" uniqKey="Brachtl G" first="G" last="Brachtl">G. Brachtl</name><affiliation wicri:level="1"><nlm:aff id="aff17"><institution>3rd Medical Department, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff18"><institution>Experimental and Clinical Cell Therapy Institute, Spinal Cord and Tissue Regeneration Center, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lin, K" sort="Lin, K" uniqKey="Lin K" first="K" last="Lin">K. Lin</name><affiliation wicri:level="1"><nlm:aff id="aff19"><institution>Royal Liverpool and Broadgreen University Hospitals NHS Trust</institution>, Liverpool,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Boysen, J" sort="Boysen, J" uniqKey="Boysen J" first="J" last="Boysen">J. Boysen</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hanson, C" sort="Hanson, C" uniqKey="Hanson C" first="C" last="Hanson">C. Hanson</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Jorgensen, J L" sort="Jorgensen, J L" uniqKey="Jorgensen J" first="J L" last="Jorgensen">J L Jorgensen</name><affiliation wicri:level="1"><nlm:aff id="aff21"><institution>MD Anderson Cancer Center</institution>, Houston, TX,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Stetler Stevenson, M" sort="Stetler Stevenson, M" uniqKey="Stetler Stevenson M" first="M" last="Stetler-Stevenson">M. Stetler-Stevenson</name><affiliation wicri:level="1"><nlm:aff id="aff22"><institution>National Cancer Institute, National Institutes of Health</institution>, Bethesda, MD,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yuan, C" sort="Yuan, C" uniqKey="Yuan C" first="C" last="Yuan">C. Yuan</name><affiliation wicri:level="1"><nlm:aff id="aff22"><institution>National Cancer Institute, National Institutes of Health</institution>, Bethesda, MD,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Broome, H E" sort="Broome, H E" uniqKey="Broome H" first="H E" last="Broome">H E Broome</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Rassenti, L" sort="Rassenti, L" uniqKey="Rassenti L" first="L" last="Rassenti">L. Rassenti</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Craig, F" sort="Craig, F" uniqKey="Craig F" first="F" last="Craig">F. Craig</name><affiliation wicri:level="1"><nlm:aff id="aff24"><institution>Un1iversity of Pittsburgh</institution>, Pittsburgh, PA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Delgado, J" sort="Delgado, J" uniqKey="Delgado J" first="J" last="Delgado">J. Delgado</name><affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Moreno, C" sort="Moreno, C" uniqKey="Moreno C" first="C" last="Moreno">C. Moreno</name><affiliation wicri:level="1"><nlm:aff id="aff5"><institution>Hospital de la Santa Creu i Sant Pau</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Bosch, F" sort="Bosch, F" uniqKey="Bosch F" first="F" last="Bosch">F. Bosch</name><affiliation wicri:level="1"><nlm:aff id="aff6"><institution>Hospital Universitari Vall d'Hebron</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Egle, A" sort="Egle, A" uniqKey="Egle A" first="A" last="Egle">A. Egle</name><affiliation wicri:level="1"><nlm:aff id="aff17"><institution>3rd Medical Department, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Doubek, M" sort="Doubek, M" uniqKey="Doubek M" first="M" last="Doubek">M. Doubek</name><affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Pospisilova, S" sort="Pospisilova, S" uniqKey="Pospisilova S" first="S" last="Pospisilova">S. Pospisilova</name><affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Mulligan, S" sort="Mulligan, S" uniqKey="Mulligan S" first="S" last="Mulligan">S. Mulligan</name><affiliation wicri:level="1"><nlm:aff id="aff25"><institution>Department of Haematology, Royal North Shore Hospital</institution>, Sydney,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Westerman, D" sort="Westerman, D" uniqKey="Westerman D" first="D" last="Westerman">D. Westerman</name><affiliation wicri:level="1"><nlm:aff id="aff14"><institution>Peter MacCallum Cancer Centre</institution>, Melbourne,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sanders, C M" sort="Sanders, C M" uniqKey="Sanders C" first="C M" last="Sanders">C M Sanders</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Emerson, R" sort="Emerson, R" uniqKey="Emerson R" first="R" last="Emerson">R. Emerson</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Robins, H S" sort="Robins, H S" uniqKey="Robins H" first="H S" last="Robins">H S Robins</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kirsch, I" sort="Kirsch, I" uniqKey="Kirsch I" first="I" last="Kirsch">I. Kirsch</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Shanafelt, T" sort="Shanafelt, T" uniqKey="Shanafelt T" first="T" last="Shanafelt">T. Shanafelt</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Pettitt, A" sort="Pettitt, A" uniqKey="Pettitt A" first="A" last="Pettitt">A. Pettitt</name><affiliation wicri:level="1"><nlm:aff id="aff19"><institution>Royal Liverpool and Broadgreen University Hospitals NHS Trust</institution>, Liverpool,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kipps, T J" sort="Kipps, T J" uniqKey="Kipps T" first="T J" last="Kipps">T J Kipps</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wierda, W G" sort="Wierda, W G" uniqKey="Wierda W" first="W G" last="Wierda">W G Wierda</name><affiliation wicri:level="1"><nlm:aff id="aff21"><institution>MD Anderson Cancer Center</institution>, Houston, TX,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Cymbalista, F" sort="Cymbalista, F" uniqKey="Cymbalista F" first="F" last="Cymbalista">F. Cymbalista</name><affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP)</institution>, Bobigny,<country>France</country></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hallek, M" sort="Hallek, M" uniqKey="Hallek M" first="M" last="Hallek">M. Hallek</name><affiliation wicri:level="1"><nlm:aff id="aff8"><institution>University Hospital</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hillmen, P" sort="Hillmen, P" uniqKey="Hillmen P" first="P" last="Hillmen">P. Hillmen</name><affiliation wicri:level="1"><nlm:aff id="aff27"><institution>Leeds Institute of Cancer and Pathology, University of Leeds</institution>, Leeds,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Montserrat, E" sort="Montserrat, E" uniqKey="Montserrat E" first="E" last="Montserrat">E. Montserrat</name><affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Ghia, P" sort="Ghia, P" uniqKey="Ghia P" first="P" last="Ghia">P. Ghia</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff28"><institution>Università Vita-Salute San Raffaele</institution>, Milan,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26639181</idno><idno type="pmc">4832072</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4832072</idno><idno type="RBID">PMC:4832072</idno><idno type="doi">10.1038/leu.2015.313</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000725</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000725</idno><idno type="wicri:Area/Pmc/Curation">000724</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000724</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study</title><author><name sortKey="Rawstron, A C" sort="Rawstron, A C" uniqKey="Rawstron A" first="A C" last="Rawstron">A C Rawstron</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>HMDS, St James's Institute of Oncology</institution>, Leeds,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Fazi, C" sort="Fazi, C" uniqKey="Fazi C" first="C" last="Fazi">C. Fazi</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Agathangelidis, A" sort="Agathangelidis, A" uniqKey="Agathangelidis A" first="A" last="Agathangelidis">A. Agathangelidis</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Villamor, N" sort="Villamor, N" uniqKey="Villamor N" first="N" last="Villamor">N. Villamor</name><affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Letestu, R" sort="Letestu, R" uniqKey="Letestu R" first="R" last="Letestu">R. Letestu</name><affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP)</institution>, Bobigny,<country>France</country></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Nomdedeu, J" sort="Nomdedeu, J" uniqKey="Nomdedeu J" first="J" last="Nomdedeu">J. Nomdedeu</name><affiliation wicri:level="1"><nlm:aff id="aff5"><institution>Hospital de la Santa Creu i Sant Pau</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Palacio, C" sort="Palacio, C" uniqKey="Palacio C" first="C" last="Palacio">C. Palacio</name><affiliation wicri:level="1"><nlm:aff id="aff6"><institution>Hospital Universitari Vall d'Hebron</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Stehlikova, O" sort="Stehlikova, O" uniqKey="Stehlikova O" first="O" last="Stehlikova">O. Stehlikova</name><affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kreuzer, K A" sort="Kreuzer, K A" uniqKey="Kreuzer K" first="K-A" last="Kreuzer">K-A Kreuzer</name><affiliation wicri:level="1"><nlm:aff id="aff8"><institution>University Hospital</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Liptrot, S" sort="Liptrot, S" uniqKey="Liptrot S" first="S" last="Liptrot">S. Liptrot</name><affiliation wicri:level="1"><nlm:aff id="aff9"><institution>St James Hospital</institution>, Dublin,<country>Ireland</country></nlm:aff><country xml:lang="fr">Irlande (pays)</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="O Brien, D" sort="O Brien, D" uniqKey="O Brien D" first="D" last="O'Brien">D. O'Brien</name><affiliation wicri:level="1"><nlm:aff id="aff9"><institution>St James Hospital</institution>, Dublin,<country>Ireland</country></nlm:aff><country xml:lang="fr">Irlande (pays)</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="De Tute, R M" sort="De Tute, R M" uniqKey="De Tute R" first="R M" last="De Tute">R M De Tute</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>HMDS, St James's Institute of Oncology</institution>, Leeds,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Marinov, I" sort="Marinov, I" uniqKey="Marinov I" first="I" last="Marinov">I. Marinov</name><affiliation wicri:level="1"><nlm:aff id="aff10"><institution>Institute of Hematology and Blood Transfusion, University Hospital Prague</institution>, Prague,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hauwel, M" sort="Hauwel, M" uniqKey="Hauwel M" first="M" last="Hauwel">M. Hauwel</name><affiliation wicri:level="1"><nlm:aff id="aff11"><institution>University Hospital Geneva</institution>, Geneva,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Spacek, M" sort="Spacek, M" uniqKey="Spacek M" first="M" last="Spacek">M. Spacek</name><affiliation wicri:level="1"><nlm:aff id="aff12"><institution>General University Hospital</institution>, Prague,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Dobber, J" sort="Dobber, J" uniqKey="Dobber J" first="J" last="Dobber">J. Dobber</name><affiliation wicri:level="1"><nlm:aff id="aff13"><institution>Department of Hematology, Academic Medical Centre</institution>, Amsterdam,<country>Netherlands</country></nlm:aff><country xml:lang="fr">Pays-Bas</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kater, A P" sort="Kater, A P" uniqKey="Kater A" first="A P" last="Kater">A P Kater</name><affiliation wicri:level="1"><nlm:aff id="aff13"><institution>Department of Hematology, Academic Medical Centre</institution>, Amsterdam,<country>Netherlands</country></nlm:aff><country xml:lang="fr">Pays-Bas</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gambell, P" sort="Gambell, P" uniqKey="Gambell P" first="P" last="Gambell">P. Gambell</name><affiliation wicri:level="1"><nlm:aff id="aff14"><institution>Peter MacCallum Cancer Centre</institution>, Melbourne,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Soosapilla, A" sort="Soosapilla, A" uniqKey="Soosapilla A" first="A" last="Soosapilla">A. Soosapilla</name><affiliation wicri:level="1"><nlm:aff id="aff15"><institution>Hematology and Flow Cytometry, Laverty Pathology</institution>, Sydney,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lozanski, G" sort="Lozanski, G" uniqKey="Lozanski G" first="G" last="Lozanski">G. Lozanski</name><affiliation wicri:level="1"><nlm:aff id="aff16"><institution>Ohio State University Medical Center</institution>, Columbus, OH,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Brachtl, G" sort="Brachtl, G" uniqKey="Brachtl G" first="G" last="Brachtl">G. Brachtl</name><affiliation wicri:level="1"><nlm:aff id="aff17"><institution>3rd Medical Department, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff18"><institution>Experimental and Clinical Cell Therapy Institute, Spinal Cord and Tissue Regeneration Center, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lin, K" sort="Lin, K" uniqKey="Lin K" first="K" last="Lin">K. Lin</name><affiliation wicri:level="1"><nlm:aff id="aff19"><institution>Royal Liverpool and Broadgreen University Hospitals NHS Trust</institution>, Liverpool,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Boysen, J" sort="Boysen, J" uniqKey="Boysen J" first="J" last="Boysen">J. Boysen</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hanson, C" sort="Hanson, C" uniqKey="Hanson C" first="C" last="Hanson">C. Hanson</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Jorgensen, J L" sort="Jorgensen, J L" uniqKey="Jorgensen J" first="J L" last="Jorgensen">J L Jorgensen</name><affiliation wicri:level="1"><nlm:aff id="aff21"><institution>MD Anderson Cancer Center</institution>, Houston, TX,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Stetler Stevenson, M" sort="Stetler Stevenson, M" uniqKey="Stetler Stevenson M" first="M" last="Stetler-Stevenson">M. Stetler-Stevenson</name><affiliation wicri:level="1"><nlm:aff id="aff22"><institution>National Cancer Institute, National Institutes of Health</institution>, Bethesda, MD,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yuan, C" sort="Yuan, C" uniqKey="Yuan C" first="C" last="Yuan">C. Yuan</name><affiliation wicri:level="1"><nlm:aff id="aff22"><institution>National Cancer Institute, National Institutes of Health</institution>, Bethesda, MD,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Broome, H E" sort="Broome, H E" uniqKey="Broome H" first="H E" last="Broome">H E Broome</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Rassenti, L" sort="Rassenti, L" uniqKey="Rassenti L" first="L" last="Rassenti">L. Rassenti</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Craig, F" sort="Craig, F" uniqKey="Craig F" first="F" last="Craig">F. Craig</name><affiliation wicri:level="1"><nlm:aff id="aff24"><institution>Un1iversity of Pittsburgh</institution>, Pittsburgh, PA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Delgado, J" sort="Delgado, J" uniqKey="Delgado J" first="J" last="Delgado">J. Delgado</name><affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Moreno, C" sort="Moreno, C" uniqKey="Moreno C" first="C" last="Moreno">C. Moreno</name><affiliation wicri:level="1"><nlm:aff id="aff5"><institution>Hospital de la Santa Creu i Sant Pau</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Bosch, F" sort="Bosch, F" uniqKey="Bosch F" first="F" last="Bosch">F. Bosch</name><affiliation wicri:level="1"><nlm:aff id="aff6"><institution>Hospital Universitari Vall d'Hebron</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Egle, A" sort="Egle, A" uniqKey="Egle A" first="A" last="Egle">A. Egle</name><affiliation wicri:level="1"><nlm:aff id="aff17"><institution>3rd Medical Department, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></nlm:aff><country xml:lang="fr">Autriche</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Doubek, M" sort="Doubek, M" uniqKey="Doubek M" first="M" last="Doubek">M. Doubek</name><affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Pospisilova, S" sort="Pospisilova, S" uniqKey="Pospisilova S" first="S" last="Pospisilova">S. Pospisilova</name><affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></nlm:aff><country xml:lang="fr">République tchèque</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Mulligan, S" sort="Mulligan, S" uniqKey="Mulligan S" first="S" last="Mulligan">S. Mulligan</name><affiliation wicri:level="1"><nlm:aff id="aff25"><institution>Department of Haematology, Royal North Shore Hospital</institution>, Sydney,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Westerman, D" sort="Westerman, D" uniqKey="Westerman D" first="D" last="Westerman">D. Westerman</name><affiliation wicri:level="1"><nlm:aff id="aff14"><institution>Peter MacCallum Cancer Centre</institution>, Melbourne,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sanders, C M" sort="Sanders, C M" uniqKey="Sanders C" first="C M" last="Sanders">C M Sanders</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Emerson, R" sort="Emerson, R" uniqKey="Emerson R" first="R" last="Emerson">R. Emerson</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Robins, H S" sort="Robins, H S" uniqKey="Robins H" first="H S" last="Robins">H S Robins</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kirsch, I" sort="Kirsch, I" uniqKey="Kirsch I" first="I" last="Kirsch">I. Kirsch</name><affiliation wicri:level="1"><nlm:aff id="aff26"><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Shanafelt, T" sort="Shanafelt, T" uniqKey="Shanafelt T" first="T" last="Shanafelt">T. Shanafelt</name><affiliation wicri:level="1"><nlm:aff id="aff20"><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Pettitt, A" sort="Pettitt, A" uniqKey="Pettitt A" first="A" last="Pettitt">A. Pettitt</name><affiliation wicri:level="1"><nlm:aff id="aff19"><institution>Royal Liverpool and Broadgreen University Hospitals NHS Trust</institution>, Liverpool,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Kipps, T J" sort="Kipps, T J" uniqKey="Kipps T" first="T J" last="Kipps">T J Kipps</name><affiliation wicri:level="1"><nlm:aff id="aff23"><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wierda, W G" sort="Wierda, W G" uniqKey="Wierda W" first="W G" last="Wierda">W G Wierda</name><affiliation wicri:level="1"><nlm:aff id="aff21"><institution>MD Anderson Cancer Center</institution>, Houston, TX,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Cymbalista, F" sort="Cymbalista, F" uniqKey="Cymbalista F" first="F" last="Cymbalista">F. Cymbalista</name><affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP)</institution>, Bobigny,<country>France</country></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hallek, M" sort="Hallek, M" uniqKey="Hallek M" first="M" last="Hallek">M. Hallek</name><affiliation wicri:level="1"><nlm:aff id="aff8"><institution>University Hospital</institution>, Cologne,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hillmen, P" sort="Hillmen, P" uniqKey="Hillmen P" first="P" last="Hillmen">P. Hillmen</name><affiliation wicri:level="1"><nlm:aff id="aff27"><institution>Leeds Institute of Cancer and Pathology, University of Leeds</institution>, Leeds,<country>UK</country></nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Montserrat, E" sort="Montserrat, E" uniqKey="Montserrat E" first="E" last="Montserrat">E. Montserrat</name><affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Ghia, P" sort="Ghia, P" uniqKey="Ghia P" first="P" last="Ghia">P. Ghia</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="aff28"><institution>Università Vita-Salute San Raffaele</institution>, Milan,<country>Italy</country></nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Leukemia</title><idno type="ISSN">0887-6924</idno><idno type="eISSN">1476-5551</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>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<sup>−5</sup>). 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<sup>−4</sup>) 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<sup>−6</sup>). 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.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Leukemia</journal-id><journal-id journal-id-type="iso-abbrev">Leukemia</journal-id><journal-title-group><journal-title>Leukemia</journal-title></journal-title-group><issn pub-type="ppub">0887-6924</issn><issn pub-type="epub">1476-5551</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26639181</article-id><article-id pub-id-type="pmc">4832072</article-id><article-id pub-id-type="pii">leu2015313</article-id><article-id pub-id-type="doi">10.1038/leu.2015.313</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL 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rid="aff25">25</xref></contrib><contrib contrib-type="author"><name><surname>Westerman</surname><given-names>D</given-names></name><xref ref-type="aff" rid="aff14">14</xref></contrib><contrib contrib-type="author"><name><surname>Sanders</surname><given-names>C M</given-names></name><xref ref-type="aff" rid="aff26">26</xref></contrib><contrib contrib-type="author"><name><surname>Emerson</surname><given-names>R</given-names></name><xref ref-type="aff" rid="aff26">26</xref></contrib><contrib contrib-type="author"><name><surname>Robins</surname><given-names>H S</given-names></name><xref ref-type="aff" rid="aff26">26</xref></contrib><contrib contrib-type="author"><name><surname>Kirsch</surname><given-names>I</given-names></name><xref ref-type="aff" rid="aff26">26</xref></contrib><contrib contrib-type="author"><name><surname>Shanafelt</surname><given-names>T</given-names></name><xref ref-type="aff" rid="aff20">20</xref></contrib><contrib contrib-type="author"><name><surname>Pettitt</surname><given-names>A</given-names></name><xref ref-type="aff" rid="aff19">19</xref></contrib><contrib contrib-type="author"><name><surname>Kipps</surname><given-names>T J</given-names></name><xref ref-type="aff" rid="aff23">23</xref></contrib><contrib contrib-type="author"><name><surname>Wierda</surname><given-names>W G</given-names></name><xref ref-type="aff" rid="aff21">21</xref></contrib><contrib contrib-type="author"><name><surname>Cymbalista</surname><given-names>F</given-names></name><xref ref-type="aff" rid="aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Hallek</surname><given-names>M</given-names></name><xref ref-type="aff" rid="aff8">8</xref></contrib><contrib contrib-type="author"><name><surname>Hillmen</surname><given-names>P</given-names></name><xref ref-type="aff" rid="aff27">27</xref></contrib><contrib contrib-type="author"><name><surname>Montserrat</surname><given-names>E</given-names></name><xref ref-type="aff" rid="aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Ghia</surname><given-names>P</given-names></name><xref ref-type="aff" rid="aff2">2</xref><xref ref-type="aff" rid="aff28">28</xref></contrib><on-behalf-of>on behalf of ERIC (European Research Initiative on CLL)</on-behalf-of><aff id="aff1"><label>1</label><institution>HMDS, St James's Institute of Oncology</institution>, Leeds,<country>UK</country></aff><aff id="aff2"><label>2</label><institution>IRCCS San Raffaele Scientific Institute</institution>, Milano,<country>Italy</country></aff><aff id="aff3"><label>3</label><institution>Hospital Clínic, IDIBAPS</institution>, Barcelona,<country>Spain</country></aff><aff id="aff4"><label>4</label><institution>Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP)</institution>, Bobigny,<country>France</country></aff><aff id="aff5"><label>5</label><institution>Hospital de la Santa Creu i Sant Pau</institution>, Barcelona,<country>Spain</country></aff><aff id="aff6"><label>6</label><institution>Hospital Universitari Vall d'Hebron</institution>, Barcelona,<country>Spain</country></aff><aff id="aff7"><label>7</label><institution>Department of Internal Medicine—Hematology and Oncology, University Hospital Brno and CEITEC, Masaryk University</institution>, Brno,<country>Czech Republic</country></aff><aff id="aff8"><label>8</label><institution>University Hospital</institution>, Cologne,<country>Germany</country></aff><aff id="aff9"><label>9</label><institution>St James Hospital</institution>, Dublin,<country>Ireland</country></aff><aff id="aff10"><label>10</label><institution>Institute of Hematology and Blood Transfusion, University Hospital Prague</institution>, Prague,<country>Czech Republic</country></aff><aff id="aff11"><label>11</label><institution>University Hospital Geneva</institution>, Geneva,<country>Switzerland</country></aff><aff id="aff12"><label>12</label><institution>General University Hospital</institution>, Prague,<country>Czech Republic</country></aff><aff id="aff13"><label>13</label><institution>Department of Hematology, Academic Medical Centre</institution>, Amsterdam,<country>Netherlands</country></aff><aff id="aff14"><label>14</label><institution>Peter MacCallum Cancer Centre</institution>, Melbourne,<country>Australia</country></aff><aff id="aff15"><label>15</label><institution>Hematology and Flow Cytometry, Laverty Pathology</institution>, Sydney,<country>Australia</country></aff><aff id="aff16"><label>16</label><institution>Ohio State University Medical Center</institution>, Columbus, OH,<country>USA</country></aff><aff id="aff17"><label>17</label><institution>3rd Medical Department, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></aff><aff id="aff18"><label>18</label><institution>Experimental and Clinical Cell Therapy Institute, Spinal Cord and Tissue Regeneration Center, Paracelsus Medical University</institution>, Salzburg,<country>Austria</country></aff><aff id="aff19"><label>19</label><institution>Royal Liverpool and Broadgreen University Hospitals NHS Trust</institution>, Liverpool,<country>UK</country></aff><aff id="aff20"><label>20</label><institution>Divisions of Hematology and Hematopathology, Mayo Clinic</institution>, Rochester, MN,<country>USA</country></aff><aff id="aff21"><label>21</label><institution>MD Anderson Cancer Center</institution>, Houston, TX,<country>USA</country></aff><aff id="aff22"><label>22</label><institution>National Cancer Institute, National Institutes of Health</institution>, Bethesda, MD,<country>USA</country></aff><aff id="aff23"><label>23</label><institution>Moores Cancer Center, University of California, San Diego</institution>, La Jolla, CA,<country>USA</country></aff><aff id="aff24"><label>24</label><institution>Un1iversity of Pittsburgh</institution>, Pittsburgh, PA,<country>USA</country></aff><aff id="aff25"><label>25</label><institution>Department of Haematology, Royal North Shore Hospital</institution>, Sydney,<country>Australia</country></aff><aff id="aff26"><label>26</label><institution>Adaptive Biotechnologies</institution>, Seattle, WA,<country>USA</country></aff><aff id="aff27"><label>27</label><institution>Leeds Institute of Cancer and Pathology, University of Leeds</institution>, Leeds,<country>UK</country></aff><aff id="aff28"><label>28</label><institution>Università Vita-Salute San Raffaele</institution>, Milan,<country>Italy</country></aff></contrib-group><author-notes><corresp id="caf1"><label>*</label><institution>HMDS, St James's Institute of Oncology, Leeds Teaching Hospitals NHS Trust</institution>, Bexley Wing, Beckett Street, Leeds LS9 7TF, <country>UK</country>. E-mail: <email>andy.rawstron@hmds.org.uk</email></corresp></author-notes><pub-date pub-type="ppub"><month>04</month><year>2016</year></pub-date><pub-date pub-type="epreprint"><day>07</day><month>12</month><year>2015</year></pub-date><pub-date pub-type="epub"><day>29</day><month>01</month><year>2016</year></pub-date><volume>30</volume><issue>4</issue><fpage>929</fpage><lpage>936</lpage><history><date date-type="received"><day>08</day><month>05</month><year>2015</year></date><date date-type="rev-recd"><day>14</day><month>08</month><year>2015</year></date><date date-type="accepted"><day>14</day><month>09</month><year>2015</year></date></history><permissions><copyright-statement>Copyright © 2016 Macmillan Publishers Limited</copyright-statement><copyright-year>2016</copyright-year><copyright-holder>Macmillan Publishers Limited</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc-nd/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc-nd/4.0/">http://creativecommons.org/licenses/by-nc-nd/4.0/</ext-link></license-p></license></permissions><abstract><p>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<sup>−5</sup>). 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<sup>−4</sup>) 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<sup>−6</sup>). 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.</p></abstract></article-meta></front><floats-group><fig id="fig1"><label>Figure 1</label><caption><p>(<bold>a</bold>) 8-CLR 1-tube panel dilution analysis. Data from serial dilution analysis of 5 × 1:10 dilutions on five CLL cases were analyzed using a single-tube eight-marker panel. Markers with a dark gray fill indicate results above the limit of quantitation; markers with a light gray fill indicate results below the limit of quantitation but above the limit of detection; and markers with no fill indicate results below the limit of detection. For log-transformed data above the LOQ, linearity=1.01, correlation coefficient Pearson <italic>R</italic>=0.99. (<bold>b</bold>) Confirmation that six markers are sufficient for detection of MRD: Bland–Altman plot comparing MRD level calculated using the single-tube eight-marker combination against the MRD level calculated using a six-marker core panel, that is, excluding CD3 and CD22. For log-transformed data above the LOQ, linearity=1.00, correlation coefficient Pearson <italic>R</italic>=1.00, average difference=−0.0026 log, 95% limit of agreement ±0.012 log. LOQ, limit of quantification.</p></caption><graphic xlink:href="leu2015313f1"></graphic></fig><fig id="fig2"><label>Figure 2</label><caption><p>(<bold>a</bold>) Six-marker core panel dilution analysis. Data from serial dilution analysis of eight CLL cases diluted into normal peripheral blood leukocytes in serial 1:10 (<italic>n</italic>=5) or 1:5 dilutions (<italic>n</italic>=3). Markers with a dark gray fill indicate results above the limit of quantitation; markers with a light gray fill indicate results below the limit of quantitation but above the limit of detection; and markers with no fill indicate results below the limit of detection. For log-transformed data above the LOQ, linearity=1.02, correlation coefficient Pearson <italic>R</italic>=1.00. (<bold>b</bold>) Acceptable interoperator variation in analysis of the six-marker core panel: analytical variation was tested using 19 operators with experience of flow cytometry but not direct experience of MRD analysis in CLL using the six-marker core panel. The results showed good concordance at the 0.010% threshold with and acceptable 95% limit of agreement of ±0.27 log for results above the limit of quantitation. For log-transformed data above the LOQ, linearity=1.02, correlation coefficient Pearson <italic>R</italic>=0.99, average difference=0.013 log, 95% limit of agreement ±0.27 log.</p></caption><graphic xlink:href="leu2015313f2"></graphic></fig><fig id="fig3"><label>Figure 3</label><caption><p>Identification of the optimal reagent specification required for MRD analysis. CLL cells and normal B cells were separately labeled with CD19 PE-Cy7 and CD19 PerCP-Cy5.5, respectively, prior to washing and mixing to create five samples which were then incubated with serial dilutions antibodies at varying concentrations (neat to 1:243, serial 1:3 dilutions). This permitted calculation of the degree to which CLL cells overlapped with normal B cells in fluorescence intensity for each marker across a range of signal intensities. The signal intensities were calculated using internal positive and negative controls and plotted against the proportion of cases with suboptimal separation of CLL cells from normal B cells, where suboptimal separation was defined as an increase in overlap of 10% or more compared with the lowest overlap for each dilution series.</p></caption><graphic xlink:href="leu2015313f3"></graphic></fig><fig id="fig4"><label>Figure 4</label><caption><p>ClonoSEQ high-throughput sequencing (HTS) shows good linearity to one CLL cell in one million leukocytes. Analysis of three CLL cases diluted into leukocytes from leucodepletion filters in serial 1:10. Each CLL clone was tagged with two sequences, one productive and one non-productive. The plot shows the CLL sequence as a percentage of nucleated genomes. Each case has a different marker shape (square for case 1, circle for case 2 and diamond for case 3) with no fill for the productive sequence data, gray fill for the non-productive sequence and black fill for the average. For log-transformed data above the limit of detection, linearity=1.12, correlation coefficient Pearson <italic>R</italic>=0.98.</p></caption><graphic xlink:href="leu2015313f4"></graphic></fig><fig id="fig5"><label>Figure 5</label><caption><p>Comparison of the six-marker core MRD flow assay with the 4-CLR 4-tube MRD flow assay and ClonoSEQ HTS. Data from serial dilution studies (<italic>n</italic>=18) and from patient samples after FCR-based therapy (<italic>n</italic>=12) were analyzed using the harmonized 4-CLR ERIC panel, the six-marker core panel and ClonoSEQ high-throughput sequencing. (<bold>a</bold>) Comparison of the six-marker core MRD flow assay with the ERIC 4-tube 4-CLR panel: For log-transformed data above the LOQ, linearity=0.99, correlation coefficient Pearson <italic>R</italic>=1.00, average difference=−0.044 log, 95% limit of agreement ±0.17 log. (<bold>b</bold>) Comparison of the six-marker core MRD flow assay with ClonoSEQ HTS: For log-transformed data above the LOQ, linearity=0.89, correlation coefficient Pearson <italic>R</italic>=0.75, average difference=−0.12 log, 95% limit of agreement ±1.3 log.</p></caption><graphic xlink:href="leu2015313f5"></graphic></fig><table-wrap id="tbl1"><label>Table 1</label><caption><title>Target values for markers used in CLL MRD analysis</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th align="left" valign="top" charoff="50"><italic>Antigen</italic></th><th align="center" valign="top" charoff="50"><italic>Typical expression in CLL</italic><xref ref-type="fn" rid="t1-fn1">a</xref></th><th colspan="2" align="center" valign="top" charoff="50"><italic>Control populations in normal peripheral blood</italic><hr></hr></th><th align="center" valign="top" charoff="50"><italic>Minimum</italic><xref ref-type="fn" rid="t1-fn2">b</xref><italic>relative fluorescence intensity (preferred)</italic></th></tr><tr><th align="left" valign="top" charoff="50"> </th><th align="center" valign="top" charoff="50"> </th><th align="center" valign="top" charoff="50"><italic>Positive</italic></th><th align="center" valign="top" charoff="50"><italic>Negative</italic></th><th align="center" valign="top" charoff="50"> </th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">CD5</td><td align="center" valign="top" charoff="50">Positive (>20%)</td><td align="center" valign="top" charoff="50">CD3+ T cells</td><td align="center" valign="top" charoff="50">CD19+ B cells</td><td align="center" valign="top" charoff="50">>30 (>65)</td></tr><tr><td align="left" valign="top" charoff="50">CD20</td><td align="center" valign="top" charoff="50">Weak</td><td align="center" valign="top" charoff="50">CD19+ B cells</td><td align="center" valign="top" charoff="50">CD3+ T-cells</td><td align="center" valign="top" charoff="50">>10 (>20)</td></tr><tr><td align="left" valign="top" charoff="50">CD43</td><td align="center" valign="top" charoff="50">Positive (>20%)</td><td align="center" valign="top" charoff="50">CD3+ T cells</td><td align="center" valign="top" charoff="50">CD20+ B cells</td><td align="center" valign="top" charoff="50">>15 (>40)</td></tr><tr><td align="left" valign="top" charoff="50">CD79b</td><td align="center" valign="top" charoff="50">Weak</td><td align="center" valign="top" charoff="50">CD20+ B cells</td><td align="center" valign="top" charoff="50">CD3+ T-cells</td><td align="center" valign="top" charoff="50">>15 (>30)</td></tr><tr><td align="left" valign="top" charoff="50">CD81</td><td align="center" valign="top" charoff="50">Weak</td><td align="center" valign="top" charoff="50">CD3+ T cells</td><td align="center" valign="top" charoff="50">Granulocytes</td><td align="center" valign="top" charoff="50">>12 (>20)</td></tr></tbody></table><table-wrap-foot><fn id="t1-fn1"><label>a</label><p>For the typical expression pattern, 'positive' indicates percentage of cells positive compared with control, and 'weak' indicates at least 20% reduction in fluorescence intensity relative to the median expression observed with a reference population of polyclonal B cells using the same antibody.</p></fn><fn id="t1-fn2"><label>b</label><p>The minimum relative fluorescence intensity would provide separation of CLL cells from normal B cells in >95% of cases (see <xref ref-type="fig" rid="fig3">Figure 3</xref>) with a preferred relative fluorescence intensity being the level at which >99% of cases have optimal separation of CLL cells from normal B cells.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tbl2"><label>Table 2</label><caption><title>Harmonized methods for residual disease detection using ERIC-harmonized approaches</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="left"></col><col align="left"></col><col align="left"></col></colgroup><thead valign="bottom"><tr><th align="left" valign="top" charoff="50"><italic>Application</italic></th><th align="left" valign="top" charoff="50"><italic>Assay</italic></th><th align="left" valign="top" charoff="50"><italic>Advantages</italic></th><th align="left" valign="top" charoff="50"><italic>Disadvantages</italic></th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">Trials aiming for disease control rather than eradication, e.g. continuous BCR pathway inhibition</td><td align="left" valign="top" charoff="50">Clonality assessment</td><td align="left" valign="top" charoff="50">•Relatively inexpensive and simple</td><td align="left" valign="top" charoff="50">•Requires capacity to reflex to full MRD assay if CLL cells <1.0% and/or B cells polyclonal</td></tr><tr><td align="left" valign="top" charoff="50">Trials focusing on achieving <0.010% MRD, i.e. with broadly similar responses rates to FCR,</td><td align="left" valign="top" charoff="50">4-color
4-tube</td><td align="left" valign="top" charoff="50">•Published outcome data.
•Does not require pretreatment phenotype for typical CLL</td><td align="left" valign="top" charoff="50">•Limit of detection >0.0050%
•More material required to achieve higher detection limits</td></tr><tr><td align="left" valign="top" charoff="50">requiring an MRD assessment that is published and has been used in previous clinical trials</td><td align="left" valign="top" charoff="50">6-color
2-tube</td><td align="left" valign="top" charoff="50">•Harmonized with 4-color assay
•Does not require pretreatment phenotype for typical CLL</td><td align="left" valign="top" charoff="50">•Intermediate LOD/LOQ
•Intermediate amount of material required to achieve higher detection limits</td></tr><tr><td align="left" valign="top" charoff="50">Trials aiming for significant improvements in disease depletion compared with FCR</td><td align="left" valign="top" charoff="50">6-color core panel for ⩾6-color assays
1-tube</td><td align="left" valign="top" charoff="50">•Permits flexibility for individual laboratory requirements
•LOD 0.0010% (10-5), LOQ 0.0025%
•Allows simultaneous analysis of additional markers</td><td align="left" valign="top" charoff="50">•Knowledge of pretreatment phenotype preferable</td></tr><tr><td align="left" valign="top" charoff="50"> </td><td align="left" valign="top" charoff="50">High throughput sequencing</td><td align="left" valign="top" charoff="50">•LOD 0.00010% (10-6)
•Objective analysis, does not necessarily require expert interpretation</td><td align="left" valign="top" charoff="50">•Further development work on standardization of the quantification</td></tr></tbody></table><table-wrap-foot><fn id="t2-fn1"><p>Abbreviations: LOD, limit of detection; LOQ, limit of quantification.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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