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Granulocytes Negatively Regulate Secretion of Transforming Growth Factor β1 by Bone Marrow Mononuclear Cells via Secretion of Erythropoietin Receptors in the Milieu

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Abstract

In my previous study, I demonstrated that bone marrow-derived mononuclear cells (BM MNCs) secrete copious amounts of Transforming Growth Factor β1 (TGFβ1) in response to erythropoietin (EPO). In this study, I investigated the principal cell type involved in the process. I found that a large percentage of various marrow cells, but not their mature counterparts present in the peripheral blood, express EPO-receptors (EPO-R). Cell depletion experiments showed that depletion of Glycophorin positive erythroblasts and CD41+ megakaryocytes – the prime suspects – did not affect the EPO-mediated TGFβ1 secretion by the BM MNCs. However, individual depletion of CD2+ T lymphocytes, CD14+ monocyte/macrophages, and CD19+ B cells affected the TGFβ1 secretion by EPO-primed MNCs: depletion of CD2+ cells had the most striking effect. Unexpectedly, and most interestingly, depletion of CD15+ granulocytes led to a significant increase in the TGFβ1 secretion by both naïve and EPO-primed BM MNCs, suggesting that these cells negatively regulate the process. Mechanistically, I show that the CD15+ cells exert this regulatory effect via secretion of both full-length and soluble EPO-R in the milieu. Overall my results, for the first time, unravel an in-built regulatory mechanism prevailing in the BM microenvironment that regulates the secretion of TGFβ1 by controlling EPO-EPO-R interaction.

My data could be relevant in understanding the pathophysiology of several conditions associated with deregulated production of TGFβ1 in the marrow compartment.

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Abbreviations

BM:

Bone marrow

PBL:

Peripheral blood-derived leukocytes

EPO:

Erythropoietin

EPO-R:

Erythropoietin Receptor

IMDM:

Iscove's Modified Dulbecco's Medium

FBS:

Fetal Bovine Serum

MNC:

Mononuclear Cells

TGFβ1:

Transforming Growth Factor beta 1

2-ME:

2-Mercaptoethanol

EMST:

EPO-mediated secretion of TGFβ1

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Acknowledgements

The author wishes to thank National Centre for Cell Science (DBT-NCCS), Pune, India, and Symbiosis Centre for Research & Innovation, Symbiosis International (Deemed University), Pune, India, for funding and providing infrastructural support, respectively; Dr. Charudatta Apte, Sahyadri Hospital, Pune, India for providing iliac crest samples; Dr. L.C. Padhy, ex-scientist, TIFR, Mumbai, India, for insightful discussions and generous sharing of reagents; and flow cytometry facility of DBT-NCCS for sample acquisition. The author also wishes to thank the anonymous reviewer for his/her excellent critique.

Funding

This study was funded by an intramural grant by National Centre for Cell Science, (DBT-NCCS), Pune, India.

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Authors and Affiliations

  1. Symbiosis Centre for Stem Cell Research (SCSCR), Symbiosis School of Biological Sciences, Symbiosis International University Symbiosis Knowledge park, Lavale, Pune, 412115, India

    Vaijayanti Kale

  2. National Centre for Cell Science, Ganeshkhind, Pune, 411007, India

    Vaijayanti Kale

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Vaijayanti Kale: Conceptualization, Methodology, Investigation, Validation, Formal Analysis and investigation, Resources, Writing – original draft preparation and Reviewing & Editing, Project Administration, Funding Acquisition.

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Correspondence to Vaijayanti Kale.

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The author has no relevant financial or non-financial interests to disclose

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The study was approved by the Institutional Ethics Committee (IEC) and Institutional Committee for Stem Cell Research (IC-SCR) of National Centre for Cell Science (NCCS), Pune and certify that the study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

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Kale, V. Granulocytes Negatively Regulate Secretion of Transforming Growth Factor β1 by Bone Marrow Mononuclear Cells via Secretion of Erythropoietin Receptors in the Milieu. Stem Cell Rev and Rep 18, 1408–1416 (2022). https://doi.org/10.1007/s12015-021-10292-x

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