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01.12.2017 | Research article | Ausgabe 1/2017 Open Access

Arthritis Research & Therapy 1/2017

Microparticles from patients with systemic lupus erythematosus induce production of reactive oxygen species and degranulation of polymorphonuclear leukocytes

Zeitschrift:
Arthritis Research & Therapy > Ausgabe 1/2017
Autoren:
Line Kjær Winberg, Søren Jacobsen, Claus H. Nielsen
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​s13075-017-1437-3) contains supplementary material, which is available to authorized users.

Abstract

Background

The interaction of circulating microparticles (MPs) with immune cells in systemic lupus erythematosus (SLE) is sparsely investigated. We examined the ability of MPs from SLE patients to induce production of reactive oxygen species (ROS) and degranulation of polymorphonuclear leukocytes (PMNs).

Methods

Plasma MPs, leukocytes and sera isolated from 20 SLE patients and 10 healthy controls were mixed in different combinations, with or without lipopolysaccharide (LPS), and incubated for 30 min. Dihydrorhodamine 123 was used to measure ROS production by flow cytometry. The ability of immunoglobulin G (IgG) isolated from five SLE patients to increase MP-induced production of ROS by PMNs was tested. Cell supernatants were analysed for content of primary, secondary and tertiary granule components by Luminex assays.

Results

MPs from SLE patients promoted ROS production by PMNs, and enhanced LPS-induced ROS production and release of primary granules by PMNs, when added to samples of autologous leukocytes and serum. In a similar autologous setting, MPs from healthy controls enhanced LPS-induced ROS production by PMNs. When leukocytes from a healthy control were stimulated with autologous MPs in the presence of various sera, SLE patient serum promoted ROS production and release of primary and secondary granules by PMNs. A role for antibodies in this respect was indicated by the observation that supplementation of normal serum with IgG from SLE patients promoted MP-induced ROS production by healthy PMNs. Moreover, when various MPs were incubated with leukocytes and serum from a healthy control, patient-derived MPs induced more ROS production by PMNs than did healthy control-derived MPs.

Conclusions

SLE patients display increased ROS production and degranulation by PMNs in response to MPs, which partly depends on serum components, including antibodies, MP properties and hyper-responsiveness of the PMNs per se.
Zusatzmaterial
Additional file 3: Is a figure showing MP-induced ROS production by PMNs in the presence of SLE sera. Leukocytes from a healthy blood group 0 donor were suspended in a medium containing 25% v/v of serum from 20 SLE patients (see Table 1, frozen samples). DHR was used as probe for H2O2, and the cells were stimulated with autologous MPs in combination with LPS for 30 min at 37 °C before flow cytometry. Correlation between the resulting median fluorescence intensity (MFI) after subtraction of background fluorescence (unstimulated cells) and levels of circulating anti-dsDNA antibodies. (JPG 175 kb)
Literatur
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