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The clinical significance of PNH-phenotype cells accounting for < 0.01% of total granulocytes detected by the Clinical and Laboratory Standards Institute methods in patients with bone marrow failure

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Abstract

Small populations of glycosylphosphatidylinositol-anchored protein-deficient (GPI[-]) cells accounting for up to 0.01% of total granulocytes can be accurately detected by a high-sensitivity flow cytometry (FCM) assay established by the Clinical and Laboratory Standards Institute (CLSI method) and have a prognostic value in bone marrow failure (BMF); however, the significance of GPI(-) granulocytes accounting for 0.001–0.009% of granulocytes remains unclear. To clarify this issue, we examined the peripheral blood of 21 BMF patients in whom minor (around 0.01%) populations of GPI(-) granulocytes had been previously detected by a different high-resolution FCM method (OPTIMA method, which defines ≥ 0.003% GPI(-) granulocytes as an abnormal increase) using both the CLSI and OPTIMA methods simultaneously. These two methods detected an “abnormal increase” in GPI(-) granulocytes in 10 patients (48%) and 17 patients (81%), respectively. CLSI detected 0.002–0.005% (median, 0.004%) GPI(-) granulocytes in 7 patients who were deemed positive for PNH-type cells according to the OPTIMA method, which detected 0.003–0.012% (median 0.006%) GPI(-) granulocytes. The clone sizes of GPI(-) cells detected by each assay were positively correlated (r = 0.994, p < 0.001). Of the seven patients who were judged positive for PNH-type cells by OPTIMA alone, five received immunosuppressive therapy, and all of them achieved a partial or complete response. GPI(-) granulocytes detected in BMF patients by the CLSI method should thus be considered significant, even at percentages of < 0.01%.

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Acknowledgments

We thank Rie Ohmi (Kanazawa University) and Takanobu Kimura in Japan PNH Study Group for their excellent technical support, and all of the participating physicians and registered patients who took part in this study.

Funding

This study received research fund from Alexion.

Author information

Authors and Affiliations

  1. Department of Hematology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, 920-8641, Japan

    Kohei Hosokawa, Ken Ishiyama, Mai Anh Thi Nguyen & Shinji Nakao

  2. Japan PNH Study Group, Tokyo, Japan

    Kohei Hosokawa, Ken Ishiyama, Toshiyuki Ikemoto, Chiharu Sugimori, Hideyoshi Noji, Tsutomu Shichishima, Naoshi Obara, Shigeru Chiba, Haruhiko Ninomiya, Mai Anh Thi Nguyen, Yukari Shirasugi, Yoshihiko Nakamura, Kiyoshi Ando, Yasutaka Ueda, Yuji Yonemura, Tatsuya Kawaguchi, Jun-ichi Nishimura, Yuzuru Kanakura & Shinji Nakao

  3. Department of Clinical Laboratory, Shiga University of Medical Science Hospital, Ohtsu, Japan

    Toshiyuki Ikemoto

  4. Department of Hematology, Ishikawa Prefectural Central Hospital, Kanazawa, Ishikawa, Japan

    Chiharu Sugimori

  5. Department of Hematology, Fukushima Medical University, Fukushima, Japan

    Hideyoshi Noji & Tsutomu Shichishima

  6. Department of Hematology, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan

    Naoshi Obara & Shigeru Chiba

  7. Department of Medical Sciences, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan

    Haruhiko Ninomiya

  8. Department of Hematology and Oncology, Tokai University, Isehara, Japan

    Yukari Shirasugi, Yoshihiko Nakamura & Kiyoshi Ando

  9. Department of Hematology and Oncology, Osaka University, Osaka, Japan

    Yasutaka Ueda, Jun-ichi Nishimura & Yuzuru Kanakura

  10. Department of Hematology and Infectious Diseases, Kumamoto University, Kumamoto, Japan

    Yuji Yonemura & Tatsuya Kawaguchi

Authors
  1. Kohei Hosokawa
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  2. Ken Ishiyama
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  3. Toshiyuki Ikemoto
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  4. Chiharu Sugimori
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  5. Hideyoshi Noji
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  6. Tsutomu Shichishima
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  7. Naoshi Obara
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  8. Shigeru Chiba
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  9. Haruhiko Ninomiya
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  10. Mai Anh Thi Nguyen
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  11. Yukari Shirasugi
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  12. Yoshihiko Nakamura
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  14. Yasutaka Ueda
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  17. Jun-ichi Nishimura
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  18. Yuzuru Kanakura
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  19. Shinji Nakao
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Contributions

KH, KI, TI, CS, HN, TS, NO, MATN, YS, YN, YU, YY, and TK performed the research; KH, KI, TS, SC, HN, KA, YY, and TK collected patients’ samples. KH, CS, JN, YK, and SN designed the study; KH and SN wrote the paper.

Corresponding author

Correspondence to Shinji Nakao.

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Conflict of interest

Dr. Ninomiya, Dr. Shichishima, Dr. Yonemura, Dr. Kawaguchi, and Dr. Nakao report personal fees from Alexion Pharma G.K. outside the submitted work; Dr. Ninomiya, Dr. Chiba, Dr. Ueda, Dr. Yonemura, and Dr. Nakao report grants from Alexion Pharma G.K. outside the submitted work; Dr. Kanakura and Dr. Nishimura report grants and personal fees from Alexion Pharma G.K. during the conduct of the study; Dr. Shirasugi reports honoraria from Novartis; Dr. Hosokawa, Dr. Ishiyama, Dr. Sugimori, Dr. Noji, Dr. Obara, Dr. Thi Nguyen, Dr. Nakamura, and Dr. Ando have nothing to declare.

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Hosokawa, K., Ishiyama, K., Ikemoto, T. et al. The clinical significance of PNH-phenotype cells accounting for < 0.01% of total granulocytes detected by the Clinical and Laboratory Standards Institute methods in patients with bone marrow failure. Ann Hematol 100, 1975–1982 (2021). https://doi.org/10.1007/s00277-020-04314-w

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  • DOI: https://doi.org/10.1007/s00277-020-04314-w

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