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Peripheral blood mononuclear cells - Can they provide a clue to the pathogenesis of Graves’ Orbitopathy?

Abstract

Purpose

Graves’ disease (GD) is an autoimmune disorder affecting primarily the thyroid gland. The most common extrathyroidal manifestation of GD is known as Graves’ orbitopathy (GO). Bone marrow-derived fibrocytes represent a subset of monocytes in peripheral blood mononuclear cells (PBMCs), infiltrate the orbital tissues, and contribute to the pathogenesis of GO. Hence objectives of the study included whether the concentration of fibrocytes in peripheral blood was higher in GO, whether TSHR m RNA expression and TSHR surface expression in peripheral blood were higher in GO in comparison to Graves’ Disease (GD) and Control subjects.

Methods

The percentage of circulating fibrocytes (FC) along with TSHR on its cell surface (CD 34+, CD 45+, CXCR4+, Collagen 1+, TSHR+) were assessed by flow cytometry of 50 patients with GD and GO and 15 healthy donors (Control). TSHR mRNA expression was measured by q RT PCR.

Result

The concentration of circulating fibrocytes was significantly higher in GO compared to GD and control [GO 17% vs GD 3% vs control 0.7% (p < 0.05)]. Moreover, these fibrocytes express a significantly higher level of TSHR in GO. This was corroborated by the measure of TSH mRNA; in GD it was 2.3-fold higher and in GO it was 3.9 fold higher than in control, in GO this transcript level was 1.7fold higher than GD (p < 0.05). TSHR+ fibrocytes were significantly positively correlated with CAS (p = 0.004) and negatively correlated with age (p = 0.01) and duration of disease (p = 0.01) in GO.

Conclusion

This study sheds further light on the pathogenesis of GO.

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Data availability

The datasets used and analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

GO:

Graves’ Orbitopathy

TSHR:

Thyroid Stimulating Hormone Receptor

GD:

Graves’ Disease

PBMC:

Peripheral Blood mononuclear cells

FC:

Fibrocytes

CAS:

Clinical Activity Score

mRNA:

Messenger Ribonucleic Acid

EUGOGO:

European Group of Graves’ Orbitopathy

MRI:

Magnetic resonance imaging

CLIA:

Chemiluminescence Immune Assay

ECLIA:

Electrochemiluminescence Immune Assay

TRAb:

Thyroid Stimulating Hormone Receptor Antibody

qRT-PCR:

quantitative Real Time Polymerase Chain Reaction

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

CI:

Confidence Interval.

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Acknowledgements

We are grateful to all the study participants and the members of our research team for their kind support and cooperation. We are thankful to Prof Mitali Chatterjee [Nodal Officer, Multi-Disciplinary Research Unit (MRU) IPGME&R] for her guidance in flowcytometry.

Author contribution

M.B.: Conceptualization, Investigation, Visualization, writing original draft, T.S.: Analysis of RT PCR data, A.K.: Clinical data acquisition from patients, P.B.: Data curation, validation, writing- review and editing, M.D.: Initial Conceptualization, Project Administration, S.C.: Conceptualization, project administration, supervision, data curation, validation, writing- review and editing.

Funding

We are also thankful to Department of Higher Education, Science Technology and Biotechnology (DHESTBT, West Bengal, Sanction No. 298 (Sanc.)/ST/P/S&T/9G-24/2016 dated on 28.03.2018) and Endocrine Society of India (Sanction No. R2017002 dated on 26.08.2017) for their financial support in this study.

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Correspondence to Subhankar Chowdhury.

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The authors declare no competing interests.

Ethics approval

The study has the approval of institutional ethics committee [Institute of Post Graduate Medical Education and Research and SSKM Hospital, Kolkata, West Bengal, India. (Memo No. Inst/IEC/2016/445)].

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Patients gave their informed written consent to participate.

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Basak, M., Sanyal, T., Kar, A. et al. Peripheral blood mononuclear cells - Can they provide a clue to the pathogenesis of Graves’ Orbitopathy?. Endocrine 75, 447–455 (2022). https://doi.org/10.1007/s12020-021-02865-0

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Keywords

  • Fibrocytes
  • peripheral blood mononuclear cells
  • TSHR
  • Graves Orbitopathy
  • Collagen1