Annals of Hematology

, Volume 96, Issue 4, pp 665–680 | Cite as

Comprehensive assessment of peripheral blood TCRβ repertoire in infectious mononucleosis and chronic active EBV infection patients

  • Shenglin Liu
  • Qian Zhang
  • Dongli Huang
  • Wenli Zhang
  • Fengluan Zhong
  • Jia Feng
  • Xueru Chen
  • Qingxiang Meng
  • Xiaofan Chen
  • Wei Zhang
  • Hongyu Zhang
Original Article

Abstract

Epstein-Barr virus (EBV) primary infection is usually asymptomatic, but it sometimes progresses to infectious mononucleosis (IM). Occasionally, some people develop chronic active EBV infection (CAEBV) with underlying immunodeficiency, which belongs to a continuous spectrum of EBV-associated lymphoproliferative disorders (EBV+ LPD) with heterogeneous clinical presentations and high mortality. It has been well established that T cell-mediated immune response plays a critical role in the disease evolution of EBV infection. Recently, high-throughput sequencing of the hypervariable complementarity-determining region 3 (CDR3) segments of the T cell receptor (T cell receptor β (TCRβ)) has emerged as a sensitive approach to assess the T cell repertoire. In this study, we fully characterized the diversity of peripheral blood TCRβ repertoire in IM (n = 6) and CAEBV patients (n = 5) and EBV-seropositive controls (n = 5). Compared with the healthy EBV-seropositive controls, both IM and CAEBV patients demonstrate a significant decrease in peripheral blood TCRβ repertoire diversity, basically, including narrowed repertoire breadth, highly expanded clones, and skewed CDR3 length distribution. However, there is no significant difference between IM and CAEBV patients. Furthermore, we observed some disease-related preferences in TRBV/TRBJ usage and combinations, as well as lots of T cell clones shared by different groups (unique or overlapped) involved in public T cell responses, which provide more detailed insights into the divergent disease evolution.

Keywords

Epstein-Barr virus (EBV) Infectious mononucleosis (IM) Chronic active EBV infection (CAEBV) T cell receptor (TCRβ) repertoire complementarity-determining region 3 (CDR3) 

Abbreviations

EBV

Epstein-Barr virus

IM

Infectious mononucleosis

CAEBV

Chronic active EBV infection

TCRβ

T cell receptor β

CDR3

Hypervariable complementarity-determining region 3

EBV+ LPD

EBV-associated lymphoproliferative disorders

CTLs

Cytotoxic T lymphocytes

Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (grant no. 81371737) and the Basic Research Program of Shenzhen Innovation Council of China (grant no. JCYJ20130402114702128).

Compliance with ethical standards

All procedures were performed according to the ethical standards of the Declaration of Helsinki involving human subjects in medical research. All participants gave written informed consent.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

277_2016_2911_Fig10_ESM.gif (27 kb)
Supplementary Figure 1.

Comparison of CDR3 length distribution in IM, CAEBV patients and controls. To the compare Gaussian fitting value (R2) between each group by Mann–Whitney U test. R2 represents the data more conform to normal distribution when closer to 1. P values less than 0.05 were considered significant. The median line and average value of each index (mean±SD) were presented. (GIF 27 kb)

277_2016_2911_MOESM1_ESM.tif (367 kb)
High resolution image (TIFF 367 kb)
277_2016_2911_Fig11_ESM.gif (99 kb)
Supplementary Figure 2.

The distribution of TRBV gene usage frequency. The X-axis depicts each TRBV gene and theY-axis depicts the percentage of each TRBV gene type. Green: controls (n=5); blue: IM (n=6); red: CAEBV (n=5). (GIF 99 kb)

277_2016_2911_MOESM2_ESM.tif (1.7 mb)
High resolution image (TIFF 1770 kb)
277_2016_2911_Fig12_ESM.gif (205 kb)
Supplementary Figure 3.

The distribution of TRBJ gene usage frequency. The X-axis depicts each TRBJ gene and the Y-axis depicts the percentage of each TRBJ subtype. Green: controls (n=5); blue: IM (n=6); red: CAEBV (n=5). (GIF 204 kb)

277_2016_2911_MOESM3_ESM.tif (2.6 mb)
High resolution image (TIFF 2701 kb)
277_2016_2911_Fig13_ESM.gif (180 kb)
Supplementary Figure 4.

The distribution of TRBV/TRBJ combination frequency. Each bubble represent aTRBV/TRBJ combination.The area of each bubble corresponds to the relative size ofTRBV-TRBJ combination frequency. Colors represent the different TRBV/TRBJ combination. (GIF 179 kb)

277_2016_2911_MOESM4_ESM.tif (4.8 mb)
High resolution image (TIFF 4910 kb)
277_2016_2911_Fig14_ESM.gif (596 kb)
Supplementary Figure 5.

Immunohistochemical staining of bone marrow (BM) or lymph node cells obtained CAEBV patients. (A) ZHY131# patient: bone marrow samples were stained with anti-CD56 antibody(positive: brown cytosol, open arrow) and were in situ hybridized of EBER(positive: purple, open arrow), respectively. (B) ZHY205# patient: bone marrow samples were stained with anti-CD3 antibody(positive: brown cytosol, open arrow) and were in situ hybridized of EBER(positive: purple, open arrow) , respectively. (C) ZHY365# patient: bone marrow samples were stained with anti-CD3 antibody(positive: brown cytosol, open arrow) and were in situ hybridized of EBER(positive: purple, open arrow) , respectively. (D) ZHY328# patient: bone marrow samples were stained with anti-CD3 antibody(positive: brown cytosol, open arrow) and were in situ hybridized of EBER(positive: purple, open arrow) , respectively. (E) ZHY328# patient: lymph node samples were double stained with anti-CD3 antibody and in situ hybridization of EBER(double positive: open arrow). (GIF 596 kb)

277_2016_2911_MOESM5_ESM.tif (8 mb)
High resolution image (TIFF 8217 kb)
277_2016_2911_MOESM6_ESM.doc (49 kb)
Supplementary Table 1 (DOC 49 kb)
277_2016_2911_MOESM7_ESM.doc (34 kb)
Supplementary Table 2 (DOC 34 kb)
277_2016_2911_MOESM8_ESM.doc (34 kb)
Supplementary Table 3 (DOC 34 kb)
277_2016_2911_MOESM9_ESM.doc (202 kb)
Supplementary Table 4 (DOC 201 kb)
277_2016_2911_MOESM10_ESM.doc (350 kb)
Supplementary Table 5 (DOC 350 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Shenglin Liu
    • 1
  • Qian Zhang
    • 2
  • Dongli Huang
    • 3
  • Wenli Zhang
    • 2
  • Fengluan Zhong
    • 2
  • Jia Feng
    • 2
  • Xueru Chen
    • 2
  • Qingxiang Meng
    • 2
  • Xiaofan Chen
    • 1
  • Wei Zhang
    • 1
  • Hongyu Zhang
    • 2
  1. 1.Biomedical Research InstituteShenzhen Peking University–The Hong Kong University of Science and Technology Medical CenterShenzhenChina
  2. 2.Department of HematologyPeking University Shenzhen HospitalShenzhenChina
  3. 3.Department of General SurgeryChangyi People’s HospitalChangyiChina

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