Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter to the Editor
  • Published:

Detection of circulating mast cells in advanced systemic mastocytosis

Leukemia volume 30, pages 1953–1956 (2016)Cite this article

Subjects

Mast cells are tissue-resident cells that express the stem cell factor receptor CD117 (KIT), the high-affinity IgE receptor (FcɛRI), and tryptase, but lack expression of CD34. Early precursors of mast cells are found in the CD34+ cell fraction in the bone marrow.1 These precursors enter the blood circulation as lineage-negative CD34hi CD117int/hi FcɛRI+ progenitors and mature when reaching the peripheral tissues.2, 3 Mast cells are not found in peripheral blood of healthy individuals, but circulating cells with a mast cell-like phenotype have been reported in systemic mastocytosis patients.4, 5, 6, 7 However, until now, it has not been clear if circulating mast cells are present only in the mast cell leukemia and aleukemic mast cell leukemia subtypes.7, 8, 9 Here, we identify, isolate and characterize circulating CD34− CD117hi FcɛRIhi mast cells in systemic mastocytosis patients with advanced disease but without any form of mast cell leukemia.

Peripheral blood from systemic mastocytosis patients (Table 1 and Supplementary Table 1) was collected in EDTA tubes. Red blood cells were lysed using ammonium chloride potassium buffer (0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA pH 7.2–7.4) or BD Pharm Lyse lysing buffer (BD Biosciences, Franklin Lakes, NJ, USA). Platelets were removed by centrifugation. The cells were stained with antibodies against CD4 (RPA-T4), CD8 (RPA-T8), CD14 (M5E2), CD19 (HIB19), CD25 (M-A251), CD34 (581), CD117 (104D2) and FcɛRIα (AER-37) (all from BD Biosciences or Biolegend, San Diego, CA, USA) in phosphate-buffered saline with 2% heat-inactivated fetal calf serum. Fluorescence-activated cell sorting and analysis were performed on a FACS Aria I, Fortessa or Canto II (BD Biosciences). At least 960 000 events were recorded per sample. A fraction of the cells was cytocentrifuged for morphological assessment, and the rest frozen and stored at −80 °C until DNA mutation analysis was performed. The thawed cells were lysed by incubation with recombinant Proteinase K (Thermo Fisher Scientific, MA, USA) for 90 min at 55 °C, followed by heat inactivation of the enzyme at 95 °C for 10 min. The KIT region was pre-amplified using Dreamtaq Polymerase (Thermo Fisher Scientific) for 50 rounds with the following oligos: 5′-GGTTTTCTTTTCTCCTCCAACC-3′ and 5′-TGCTGTGACCTTCAATGACC-3′ (TAG Copenhagen, Denmark). The pre-amplified DNA was purified using the MinElute PCR Purification Kit (Qiagen, Hilden, Germany). Samples were analyzed for the c.2447A>T mutation by quantitative real-time PCR as previously described10 and the results were confirmed using Sanger sequencing. Cord blood-derived mast cells were used as negative controls, and the cell line HMC-1.2 was used as positive control for the mutation assay. The study was approved by the local Ethics Committee of Stockholm. All participants in the study gave informed consent, documented in their medical records.

Table 1 Demographics of systemic mastocytosis patients
Full size table

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1

References

  1. Kirshenbaum AS, Goff JP, Semere T, Foster B, Scott LM, Metcalfe DD . Demonstration that human mast cells arise from a progenitor cell population that is CD34+, c-kit+, and expresses aminopeptidase N (CD13). Blood 1999; 94: 2333–2342.

    CAS  PubMed  Google Scholar 

  2. Dahlin JS, Hallgren J . Mast cell progenitors: origin, development and migration to tissues. Mol Immunol 2015; 63: 9–17.

    Article  CAS  Google Scholar 

  3. Dahlin JS, Malinovschi A, Öhrvik H, Sandelin M, Janson C, Alving K et al. Lin− CD34hi CD117int/hi FcɛRI+ cells in human blood constitute a rare population of mast cell progenitors. Blood 2016; 127: 383–391.

    Article  CAS  Google Scholar 

  4. Travis WD, Li CY, Hoagland HC, Travis LB, Banks PM . Mast cell leukemia: report of a case and review of the literature. Mayo Clin Proc 1986; 61: 957–966.

    Article  CAS  Google Scholar 

  5. Castells MC, Friend DS, Bunnell CA, Hu X, Kraus M, Osteen RT et al. The presence of membrane-bound stem cell factor on highly immature nonmetachromatic mast cells in the peripheral blood of a patient with aggressive systemic mastocytosis. J Allergy Clin Immunol 1996; 98: 831–840.

    Article  CAS  Google Scholar 

  6. Georgin-Lavialle S, Lhermitte L, Baude C, Barete S, Bruneau J, Launay JM et al. Blood CD34−c-Kit+ cell rate correlates with aggressive forms of systemic mastocytosis and behaves like a mast cell precursor. Blood 2011; 118: 5246–5249.

    Article  CAS  Google Scholar 

  7. Pardanani A, Lasho T, Chen D, Kimlinger TK, Finke C, Zblewski D et al. Aberrant expression of CD123 (interleukin-3 receptor-alpha) on neoplastic mast cells. Leukemia 2015; 29: 1605–1608.

    Article  CAS  Google Scholar 

  8. Valent P, Horny HP, Escribano L, Longley BJ, Li CY, Schwartz LB et al. Diagnostic criteria and classification of mastocytosis: a consensus proposal. Leuk Res 2001; 25: 603–625.

    Article  CAS  Google Scholar 

  9. Arredondo AR, Gotlib J, Shier L, Medeiros B, Wong K, Cherry A et al. Myelomastocytic leukemia versus mast cell leukemia versus systemic mastocytosis associated with acute myeloid leukemia: a diagnostic challenge. Am J Hematol 2010; 85: 600–606.

    Article  Google Scholar 

  10. Kristensen T, Vestergaard H, Moller MB . Improved detection of the KIT D816V mutation in patients with systemic mastocytosis using a quantitative and highly sensitive real-time qPCR assay. J Mol Diagn 2011; 13: 180–188.

    Article  CAS  Google Scholar 

  11. Sperr WR, Escribano L, Jordan JH, Schernthaner GH, Kundi M, Horny HP et al. Morphologic properties of neoplastic mast cells: delineation of stages of maturation and implication for cytological grading of mastocytosis. Leuk Res 2001; 25: 529–536.

    Article  CAS  Google Scholar 

  12. Escribano L, Orfao A, Diaz-Agustin B, Villarrubia J, Cervero C, Lopez A et al. Indolent systemic mast cell disease in adults: immunophenotypic characterization of bone marrow mast cells and its diagnostic implications. Blood 1998; 91: 2731–2736.

    CAS  PubMed  Google Scholar 

  13. Akin C, Kirshenbaum AS, Semere T, Worobec AS, Scott LM, Metcalfe DD . Analysis of the surface expression of c-kit and occurrence of the c-kit Asp816Val activating mutation in T cells, B cells, and myelomonocytic cells in patients with mastocytosis. Exp Hematol 2000; 28: 140–147.

    Article  CAS  Google Scholar 

  14. Torrey E, Simpson K, Wilbur S, Munoz P, Skikne B . Malignant mastocytosis with circulating mast cells. Am J Hematol 1990; 34: 283–286.

    Article  CAS  Google Scholar 

  15. Valent P, Sotlar K, Sperr WR, Escribano L, Yavuz S, Reiter A et al. Refined diagnostic criteria and classification of mast cell leukemia (MCL) and myelomastocytic leukemia (MML): a consensus proposal. Ann Oncol 2014; 25: 1691–1700.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Swedish Research Council—Medicine and Health, the Swedish Cancer Society, the Cancer and Allergy Foundation, Ollie and Elof Ericsson’s Foundation, Hans von Kantzow’s Foundation, the Swedish Medical Society, Åke Wiberg Foundation, Marianne and Marcus Wallenberg Foundation, the Konsul Th C. Bergh Foundation, Stockholm Cancer Association and through the regional agreement on medical training and clinical research (ALF) between Stockholm County Council and Karolinska Institutet.

Author contributions

GN, JSU, JSD and JG designed the study. JSD and JG performed the experiments. GN, JSU, JSD and JG analyzed the data. JSU, BS, TG and HH diagnosed the patients, contributed to sample collection and critically reviewed the manuscript.

Author information

Authors and Affiliations

  1. Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

    J S Dahlin, J Grootens, T Gülen & G Nilsson

  2. Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden

    J S Ungerstedt

  3. Hematology Center, Karolinska University Hospital, Stockholm, Sweden

    J S Ungerstedt

  4. Department of Laboratory Medicine, Division of Pathology, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

    B Sander

  5. Department of Respiratory Diseases and Allergy, Karolinska University Hospital Huddinge, Stockholm, Sweden

    T Gülen

  6. Department of Medical Sciences, Uppsala University, Uppsala, Sweden

    H Hägglund

  7. Section of Hematology, Uppsala University Hospital, Uppsala, Sweden

    H Hägglund

Authors
  1. J S Dahlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J S Ungerstedt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Grootens
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B Sander
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T Gülen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H Hägglund
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. G Nilsson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G Nilsson.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Leukemia website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dahlin, J., Ungerstedt, J., Grootens, J. et al. Detection of circulating mast cells in advanced systemic mastocytosis. Leukemia 30, 1953–1956 (2016). https://doi.org/10.1038/leu.2016.105

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2016.105

Search

Advanced search

Quick links