NR4A Expression by Human Marginal Zone B-Cells.

Kim Doyon-Laliberté,Matheus Aranguren,Michelle Byrns,Michel Roger,John Stagg,Meriam Memmi,Josiane Chagnon-Choquet,Johanne Poudrier,Pavel Chrobak

doi:10.3390/antib8040050

Kim Doyon-Laliberté, Matheus Aranguren + Show 7 more

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https://doi.org/10.3390/antib8040050

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Abstract

We have previously characterized a human blood CD19+CD1c+IgM+CD27+CD21loCD10+ innate-like B-cell population, which presents features shared by both transitional immature and marginal zone (MZ) B-cells, named herein “precursor-like” MZ B-cells. B-cells with similar attributes have been associated with regulatory potential (Breg). In order to clarify this issue and better characterize this population, we have proceeded to RNA-Seq transcriptome profiling of mature MZ and precursor-like MZ B-cells taken from the blood of healthy donors. We report that ex vivo mature MZ and precursor-like MZ B-cells express transcripts for the immunoregulatory marker CD83 and nuclear receptors NR4A1, 2, and 3, known to be associated with T-cell regulatory (Treg) maintenance and function. Breg associated markers such as CD39 and CD73 were also expressed by both populations. We also show that human blood and tonsillar precursor-like MZ B-cells were the main B-cell population to express elevated levels of CD83 and NR4A1-3 proteins ex vivo and without stimulation. Sorted tonsillar precursor-like MZ B-cells exerted regulatory activity on autologous activated CD4+ T-cells, and this was affected by a CD83 blocking reagent. We believe these observations shed light on the Breg potential of MZ populations, and identify NR4A1-3 as potential Breg markers, which as for Tregs, may be involved in stabilization of a regulatory status. Since expression and activity of these molecules can be modulated therapeutically, our findings may be useful in strategies aiming at modulation of Breg responses.

Highlights

We have previously reported a relatively small population (1%–2%) of B-cells expressing the surface markers CD19+ CD1c+ IgM+ CD27+ CD21lo CD10+ that is present in the blood of healthy human individuals [1,2,3]
We report that the been associated with regulatory potential (Breg) associated CD39 and CD73 enzymes [12,13] are co-expressed by both populations in human blood and tonsils
For means of NR4A1 protein detection by flow-cytometry and because of experimental suitability, we have used the PE-conjugated human REA clonal antibody directed against murine NR4A1, which cross-reacts with human NR4A1

Summary

Introduction

We have previously reported a relatively small population (1%–2%) of B-cells expressing the surface markers CD19+ CD1c+ IgM+ CD27+ CD21lo CD10+ that is present in the blood of healthy human individuals [1,2,3]. These B-cells harbored markers of transitional immature and of innate marginal zone (MZ) B-cells, and were named “precursor-like” MZ B-cells. We report gene expression of nuclear receptors NR4A1, 2, and 3 as well as the immunoregulatory surface molecule CD83 [9,10], which NR4As can directly regulate [11], by both mature MZ and precursor-like MZ B-cell populations from ex vivo human blood. We believe these observations shed light on the Breg potential of MZ populations, and identify NR4A1-3 as potential Breg markers, which as for Tregs, may be involved in stabilization of a regulatory status

Ethics Statement

Functional Assays

Statistical Analyses

Results and Discussion

Reduced following co-culture withwith precursor-like

Conclusions