Abstract
Within the last 25 years, flow cytometry and fluorescence-activated cell sorting have emerged as both routine diagnostic tools in clinical medicine and as advanced analytic tools critical in performing scientific research. This chapter aims at summarizing the use of flow cytometry in benign and malignant hematology and the monitoring of inherited and acquired immunodeficiency states. Numerous figures are provided from our laboratories at Massachusetts General Hospital that illustrate examples of these conditions. The chapter also describes novel flow cytometry-based imaging techniques, the combination of flow cytometry and mass spectrography, new software tools, and some future directions and applications of advanced instrumentation for flow cytometry.
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References
Spitzer TR, Dey BR, Chen YB, Attar E, Ballen KK (2012) The expanding frontier of hematopoietic cell transplantation. Cytometry B Clin Cytom 82:271–279
Preffer FI (2012) Issue highlights. Cytometry B Clin Cytom 82:343–344, November 2012
Rosko A, Lazarus HM (2012) Refining hematopoietic cell transplant: a concise review. Cytometry B Clin Cytom 82:266–267
Krause DS, Scadden DT, Preffer FI (2013) The hematopoietic stem cell niche—home for friend and foe? Cytometry B Clin Cytom 84: 7–20
Herbert DJ, Miller DT, Bruce Bagwell C (2012) Automated analysis of flow cytometric data for CD34+ stem cell enumeration using a probability state model. Cytometry B Clin Cytom 82:313–318
Whitby A, Whitby L, Fletcher M et al (2012) ISHAGE protocol: are we doing it correctly? Cytometry B Clin Cytom 82:9–17
Schnizlein-Bick CT, Spritzler J, Wilkening CL, Nicholson JKA, O’Gorman MRG (2000) Evaluation of TruCount absolute-count tubes for determining CD4 and CD8 cell numbers in human immunodeficiency virus-positive adults. Clin Diagn Lab Immunol 7:336–343
Warnatz K, Schlesier M (2008) Flow cytometric phenotyping of common variable immunodeficiency. Cytometry B Clin Cytom 74:261–271
van de Loosdrecht AA, Ireland R, Kern W et al (2013) Rationale for the clinical application of flow cytometry in patients with myelodysplastic syndromes: position paper of an International Consortium and the European LeukemiaNet Working Group. Leuk Lymphoma 54(3): 472–475
Bellos F, Alpermann T, Gouberman E et al (2012) Evaluation of flow cytometric assessment of myeloid nuclear differentiation antigen expression as a diagnostic marker for myelodysplastic syndromes in a series of 269 patients. Cytometry B Clin Cytom 82:295–304
Monaghan SA, Surti U, Doty K, Craig FE (2012) Altered neutrophil maturation patterns that limit identification of myelodysplastic syndromes. Cytometry B Clin Cytom 82:217–228
Della Porta MG, Lanza F, Del Vecchio L (2011) Flow cytometry immunophenotyping for the evaluation of bone marrow dysplasia. Cytometry B Clin Cytom 80:201–211
Kern W, Bacher U, Schnittger S, Alpermann T, Haferlach C, Haferlach T (2013) Multiparameter flow cytometry reveals myelodysplasia-related aberrant antigen expression in myelodysplastic/myeloproliferative neoplasms. Cytometry B Clin Cytom 84(3):194–197. doi:10.1002/cyto.b.21068
Sandes AF, Kerbauy DMB, Matarraz S, Chauffaille MD, López A, Orfao A, Yamamoto M (2013) Combined flow cytometric assessment of CD45, HLA-DR, CD34, and CD117 expression is a useful approach for reliable quantification of blast cells in myelodysplastic syndromes. Cytometry B Clin Cytom. doi:10.1002/cyto.b.21087
Swerdlow S, Campo E, Lee Harris N et al (2008) WHO classification of tumors of haematopoietic and lymphoid tissues, 4th edn. IARC, Lyon
Hevessy Z, Hudak R, Kiss-Sziraki V et al (2011) Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay. Clin Chem Lab Med 50:689–692
Lapidot T, Sirard C, Vormoor J et al (1994) A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature 367:645–648
Bonnet D, Dick JE (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 3:730–737
Wang JCY, Lapidot T, Cashman JD et al (1998) High level engraftment of NOD/SCID mice by primitive normal and leukemic hematopoietic cells from patients with chronic myeloid leukemia in chronic phase. Blood 91: 2406–2414
Eisterer W, Jiang X, Christ O et al (2005) Different subsets of primary chronic myeloid leukemia stem cells engraft immunodeficient mice and produce a model of the human disease. Leukemia 19:435–441
Castor A, Nilsson L, Astrand-Grundstrom I et al (2005) Distinct patterns of hematopoietic stem cell involvement in acute lymphoblastic leukemia. Nat Med 11:630–637
Medd PG, Clark N, Leyden K et al (2011) A novel scoring system combining expression of CD23, CD20, and CD38 with platelet count predicts for the presence of the t(11;14) translocation of mantle cell lymphoma. Cytometry B Clin Cytom 80:230–237
Campbell SM, Peters SB, Zirwas MJ, Wong HK (2010) Immunophenotypic diagnosis of primary cutaneous lymphomas. J Clin Aesthet Dermatol 3:21–25
Fromm JR (2011) Flow cytometric analysis of CD123 is useful for immunophenotyping classical Hodgkin lymphoma. Cytometry B Clin Cytom 80:91–99
Cannizzo E, Bellio E, Sohani AR et al (2010) Multiparameter immunophenotyping by flow cytometry in multiple myeloma: the diagnostic utility of defining ranges of normal antigenic expression in comparison to histology. Cytometry B Clin Cytom 78:231–238
Paiva B, Almeida J, Pérez-Andrés M et al (2010) Utility of flow cytometry immunophenotyping in multiple myeloma and other clonal plasma cell-related disorders. Cytometry B Clin Cytom 78:239–252
Frébet E, Abraham J, Geneviève F et al (2011) A GEIL flow cytometry consensus proposal for quantification of plasma cells: application to differential diagnosis between MGUS and myeloma. Cytometry B Clin Cytom 80:176–185
Drain S, Catherwood MA, Bjourson AJ, Drake MB, Kettle PJ, Alexander HD (2012) Neither P-gp SNP variants, P-gp expression nor functional P-gp activity predicts MDR in a preliminary study of plasma cell myeloma. Cytometry B Clin Cytom 82:229–237
Peceliunas V, Janiulioniene A, Matuzeviciene R, Griskevicius L (2011) Six color flow cytometry detects plasma cells expressing aberrant immunophenotype in bone marrow of healthy donors. Cytometry B Clin Cytom 80:318–323
Sutherland DR, Keeney M, Illingworth A (2012) Practical guidelines for the high-sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry. Cytometry B Clin Cytom 82: 195–208
Marinov I, Kohoutova M, Tkacova V et al (2013) Intra- and interlaboratory variability of paroxysmal nocturnal hemoglobinuria testing by flow cytometry following the 2012 Practical Guidelines for high-sensitivity paroxysmal nocturnal hemoglobinuria testing. Cytometry B Clin Cytom 84(4):229–236
Miller DT, Hunsberger BC, Bagwell CB (2012) Automated analysis of GPI-deficient leukocyte flow cytometric data using GenStoneTM. Cytometry B Clin Cytom 82:319–324
Dalal BI, Khare NS (2013) Flow cytometric testing for paroxysmal nocturnal hemoglobinuria: CD64 is better for gating monocytes than CD33. Cytometry B Clin Cytom 84:33–36
Wong L, Davis BH (2013) Monochromatic gating method by flow cytometry for high purity monocyte analysis. Cytometry B Clin Cytom 84:119–124
Borowitz MJ, Craig FE, Digiuseppe JA et al (2010) Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytometry B Clin Cytom 78:211–230
El-Nahas HA, Salem DA, El-Henawy AA, El-Nimr HI, Abdel-Ghaffar HA, El-Meadawy HA (2013) Giardia diagnostic methods in human fecal samples: a comparative study. Cytometry B Clin Cytom 84:44–49
Pastoret C, Le Priol J, Fest T, Roussel M (2013) Evaluation of FMH QuikQuant for the detection and quantification of fetomaternal hemorrhage. Cytometry B Clin Cytom 84: 37–43
Song JY, Filie AC, Venzon D, Stetler-Stevenson M, Yuan CM (2012) Flow cytometry increases the sensitivity of detection of leukemia and lymphoma cells in bronchoalveolar lavage specimens. Cytometry B Clin Cytom 82: 305–312
Bozzi F, Conca E, Manenti G, Negri T et al (2011) High CD133 expression levels in gastrointestinal stromal tumors. Cytometry B Clin Cytom 80:238–247
Stacchini A, Demurtas A, Aliberti S (2012) Flow cytometric detection of liposomal cytarabine in cerebrospinal fluid of patients treated with intrathecal chemotherapy. Cytometry B Clin Cytom 82:280–282
Preffer FI, Dombkowski D (2009) Advances in complex multiparameter flow cytometry technology: applications in stem cell research. Cytometry B Clin Cytom 76:295–314
Bodenmiller B, Zunder ER, Finck R et al (2012) Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators. Nat Biotechnol 30:858–867
Bandura DR, Baranov VI, Ornatsky OI et al (2009) Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry. Anal Chem 81:6813–6822
Bendall SC, Simonds EF, Qiu P et al (2010) Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 332:687–696
Wang L, Abbasi F, Ornatsky O et al (2012) Human CD4+ lymphocytes for antigen quantification: characterization using conventional flow cytometry and mass cytometry. Cytometry 81:567–575
Samsel L, Dagur P, Raghavachari N, Seamon C, Kato G, McCoy JP Jr (2013) Flow cytometry for morphologic and phenotypic characterization of rare circulating endothelial cells. Cytometry B Clin Cytom (in press). doi: 10.1002/cyto.b.21088
Grimwade L, Gudgin E, Bloxham D, Scott MA, Erber WN (2011) PML protein analysis using imaging flow cytometry. J Clin Pathol 64:447–450
Mirabelli P, Scalia G, Pascariello C et al (2012) ImageStream promyelocytic leukemia protein immunolocalization: in search of promyelocytic leukemia cells. Cytometry 81:232–237
Acknowledgements
The authors would like to thank Laura Dillon, David Dombkowski, Abby Kelliher, and Scott Mordecai for their gracious help and specific contributions to this chapter.
Disclaimers The authors have no conflicts of interest to disclaim.
Source of support: K08 CA138916-02 to DSK and grants 1S10OD012027-01A1 and 1S10RR020936-01 to FIP.
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Krause, D.S., DeLelys, M.E., Preffer, F.I. (2014). Flow Cytometry for Hematopoietic Cells. In: Beksaç, M. (eds) Bone Marrow and Stem Cell Transplantation. Methods in Molecular Biology, vol 1109. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-9437-9_2
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DOI: https://doi.org/10.1007/978-1-4614-9437-9_2
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