Plasmacytoid dendritic cells and RNA-containing immune complexes drive expansion of peripheral B cell subsets with an SLE-like phenotype

PLoS One. 2017 Aug 28;12(8):e0183946. doi: 10.1371/journal.pone.0183946. eCollection 2017.

Abstract

Background: Hyperactive B cells and a continuous interferon (IFN)-α production by plasmacytoid dendritic cells (pDCs) play a key role in systemic lupus erythematosus (SLE). We asked whether the interaction between B cells and pDCs stimulated with RNA-containing immune complexes affects peripheral B cell subsets.

Methods: B cells and pDCs were isolated from blood of healthy individuals and stimulated with immune complexes consisting of SLE-IgG and U1snRNP (RNA-IC). Expression of cell surface molecules as well as IL-6 and IL-10 production were determined by flow cytometry and immunoassays. Gene expression profiles were determined by a NanoString nCounter expression array.

Results: We found a remarkable increase of double negative CD27-IgD- B cells, from 7% within fresh CD19+ B cells to 37% in the RNA-IC-stimulated co-cultures of B cells and pDCs, comparable to the frequency of double negative B cells in SLE patients. Gene expression analysis of the double negative CD27-IgD- and the CD27+IgD- memory B cells revealed that twenty-one genes were differentially expressed between the two B cell subsets (≥ 2-fold, p<0.001). The, IL21R, IL4R, CCL4, CCL3, CD83 and the IKAROS Family Zinc Finger 2 (IKZ2) showed higher expression in the double negative CD27-IgD- B cells.

Conclusion: The interactions between B cells and pDCs together with RNA-containing IC led to an expansion of B cells with similar phenotype as seen in SLE, suggesting that the pDC-B cell crosstalk contributes to the autoimmune feed-forward loop in SLE.

MeSH terms

  • Antigen-Antibody Complex / immunology*
  • Antigens, CD / immunology
  • B-Lymphocyte Subsets
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / metabolism
  • Coculture Techniques
  • Dendritic Cells / immunology*
  • Gene Expression Profiling
  • HeLa Cells
  • Humans
  • Immunologic Memory
  • Interleukin-10 / biosynthesis
  • Interleukin-6 / biosynthesis
  • Lupus Erythematosus, Systemic / immunology*
  • Phenotype
  • RNA / immunology*

Substances

  • Antigen-Antibody Complex
  • Antigens, CD
  • Interleukin-6
  • Interleukin-10
  • RNA

Grants and funding

This work was supported by The Swedish Rheumatism Association (https://www.reumatikerforbundet.org), R-569291 (MLE), R-653711 (LR); King Gustaf V's 80-years Foundation, FAI-2015-0140 (MLE) (LR); Agnes och Mac Rudbergs Stiftelse (OB); and The Swedish Research Council 521-2013-2830 (LR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.