Differential expression of TNFR1 (CD120a) and TNFR2 (CD120b) on subpopulations of human monocytes

Daniëlle Hijdra,Jan C Grutters,Anke Me Claessen,Adriane Dm Vorselaars,Ger T Rijkers

doi:10.1186/1476-9255-9-38

Abstract

BackgroundThree subpopulations of monocytes can be distinguished in human blood: classical (CD14++CD16−), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++). CD16 expressing monocytes are expanded in patients with sarcoidosis and in various other inflammatory diseases. In sarcoidosis, it is unclear whether either intermediate, nonclassical or both CD16 expressing monocytes are responsible for this increase. Data relating to the monocyte subpopulations is receiving increasing attention, but the expression of TNF receptors on these subpopulations has not been studied thus far. The aim of this study was to determine frequencies of monocyte subpopulations and their expression of TNFR1 and TNFR2 in both sarcoidosis patients and healthy controls.MethodsPeripheral blood cells of sarcoidosis patients and healthy controls were stained for the markers HLA-DR, CD14, CD16, CD120a and CD120b. Cells were measured on a FACSCalibur and analyzed with FlowJo. We used Student’s t-test and a parametric One-way ANOVA for statistical analysis.ResultsSarcoidosis patients had a significant higher frequency of intermediate monocytes than healthy controls. Significant differences in TNF receptor expression were found between the monocyte subpopulations, both in sarcoidosis patients as well as in healthy controls: intermediates expressed more TNFR1 than classicals and nonclassicals and nonclassicals expressed more TNFR2 than intermediates, whereas intermediates showed higher expression than classicals.ConclusionsIn both sarcoidosis patients and healthy controls intermediate monocytes show the highest expression level of TNFR1 among monocyte subpopulations and nonclassical monocytes show the highest expression level of TNFR2. These findings, as wells as the higher frequency of intermediate monocytes in sarcoidosis patients, provide evidence for the existence of two functionally-distinct CD16 expressing monocyte subpopulations.

Highlights

Three subpopulations of monocytes can be distinguished in human blood: classical (CD14++CD16−), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++)
Monocytes subpopulations The frequency of classical, intermediate, and nonclassical monocyte subpopulations was analyzed in 38 sarcoidosis patients and 13 healthy controls, based on HLA-DR, CD14 and CD16 expression pattern
Classical monocytes accounted for 60–90% of Side scatter (SSC) x HLA-DR+ gated monocytes, intermediate monocytes for about 2–5% and nonclassical monocytes for 5–25%, both in sarcoidosis patients as well as in healthy controls

Summary

Introduction

Three subpopulations of monocytes can be distinguished in human blood: classical (CD14++CD16−), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++). An increase in the relative number of CD16 expressing monocytes was found in newly diagnosed sarcoidosis patients, suggesting an activated state of the monocytes This activated state might be an indicator for disease activity [6,7]. These investigations have not discriminated between the two CD16 expressing monocyte subpopulations It has been shown in patients with rheumatoid arthritis [8] and severe asthma [9] that intermediate monocytes are responsible for the increase in CD16 expressing monocytes. This has not been investigated in patients with sarcoidosis

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