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Recruitment of CD8+ T cells into bone marrow might explain the suppression of megakaryocyte apoptosis through high expression of CX3CR1+ in prolonged isolated thrombocytopenia after allogeneic hematopoietic stem cell transplantation

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Abstract

Prolonged isolated thrombocytopenia is a common complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT), which is associated with a poor prognosis. This study aimed to investigate the pathogenesis of prolonged isolated thrombocytopenia (PT). We analysed the expression of CX3CR1 on CD4 and CD8 T cells in bone marrow (BM) and peripheral blood (PB) at +90 days from allo-HSCT recipients with or without PT by flow cytometry analyses. We then determined the megakaryocytes ploidy distributions, apoptosis rate and Fas expression of recipients with or without PT in vitro directly or after depleting CD8+ T cells or adding purified autologous CD8+ T cells to CD8+ T-dep MNCs. We found that the percentage of CD8+ T cells in BM was higher in the patients with PT than in the controls. The elevated expression of the CX3CR1 was associated with PT. There was a marked increase in the percentage of low ploidy megakaryocytes in the recipients with PT. The depletion of CD8+ T cells increased the apoptosis of megakaryocytes and decreased the expression of Fas, which could be corrected by re-adding purified autologous CD8+ T cells. The increase of CD8+ T cells and CD8+/CX3CR1+ T cells in BM at +90 days were independent risk factors for PT according to multivariate analysis. Our data implied that the recruitment of CD8+ T cells into BM might explain the suppression of megakaryocyte apoptosis through the elevated expression of CX3CR1+ in PT after allo-HSCT. CX3CR1 might be a novel treatment target in recipients with PT.

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Acknowledgments

The authors thank Drs. Ming Hou and Jun Peng at the Haematological Oncology Centre at Qilu Hospital of Shandong University in China for their excellent technical support and Dr. X. Long Zheng at the Department of Pathology and Laboratory Medicine at The Children’s Hospital of Philadelphia and The University of Pennsylvania for his critical reading of the manuscript.

This work was supported by The National Key Technology Support Program (No. 2012BAI38B03), National Natural Science Foundation of China (No. 81270643 and No. 81470343) and Seeding Grant for Medicine and Engineering Sciences of Peking University (No. BMU20140389). This work was also supported in part by Beijing Natural Science Foundation (No. 7132194) and Specialized Research Fund for the Doctoral Program of Higher Education (No. 20120001110026). All these funding are received by Xiao-Hui Zhang. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

Authorship

Contribution: XJH conceived of and designed the experiments, reviewed the data analysis and reviewed the manuscript; XHZ helped with the design of the study and wrote the manuscript; GXW performed the laboratory testing, performed the statistical analyses and reviewed the manuscript; YRL, LPX and KYL helped with the design of the study and reviewed the manuscript; HHZ, WH, HC, YHC, FR W, JZ W, YC and Y W contributed the patient information and reviewed the manuscript; RF, HXF, MW, ML, YZ and TZ participated in collecting and analysing the data.

Conflict of interest

The authors report no potential conflicts of interest.

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Authors and Affiliations

  1. Peking University People’s Hospital, Peking University Institute of Hematology, Beijing, 100044, China

    Xiao-Hui Zhang, Guo-Xiang Wang, Hong-Hu Zhu, Yan-Rong Liu, Lan-Ping Xu, Wei Han, Huan Chen, Yu-Hong Chen, Feng-Rong Wang, Jing-Zhi Wang, Yu Wang, Ting Zhao, Yao Chen, Ru Feng, Hai-Xia Fu, Min Wang, Yi Zhou, Meng Lv, Kai-Yan Liu & Xiao-Jun Huang

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  1. Xiao-Hui Zhang
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Correspondence to Xiao-Hui Zhang.

Additional information

Xiao-Hui Zhang and Guo-Xiang Wang share the first authorship.

Xiao-Hui Zhang and Xiao-Jun Huang share the co-corresponding author.

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Supplementary Fig. 1

Flow cytometric analysis of lymphocytes from PT or controls. Flow cytometric analysis of lymphocytes from the BM and PB of the recipients with PT (n = 89) or controls (n = 94). CD3+ T cells in the PB (A) of the recipients with PT or controls; CD19+ B cells in the BM (B) and PB (C) of recipients with PT or controls. The results are presented as a percentage and expressed as the mean ± SD. NS, not significant (DOC 229 kb)

Supplementary Fig. 2

Flow cytometric analysis of T lymphocytes from PB of PT or controls. Flow cytometric analysis of T lymphocytes from the PB of the recipients with PT (n = 89) or the controls (n = 94). The results are presented as a percentage and expressed as the mean ± SD. NS, not significant (DOC 157 kb)

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Zhang, XH., Wang, GX., Zhu, HH. et al. Recruitment of CD8+ T cells into bone marrow might explain the suppression of megakaryocyte apoptosis through high expression of CX3CR1+ in prolonged isolated thrombocytopenia after allogeneic hematopoietic stem cell transplantation. Ann Hematol 94, 1689–1698 (2015). https://doi.org/10.1007/s00277-015-2436-6

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  • DOI: https://doi.org/10.1007/s00277-015-2436-6

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