Investigating the impact of Echinococcus multilocularis vesicular fluid on human cells from healthy blood donors

J Immunol Methods. 2015 Feb:417:52-59. doi: 10.1016/j.jim.2014.12.006. Epub 2014 Dec 13.

Abstract

Context and objectives: Alveolar echinococcosis (AE) is a severe chronic helminthic disease that mimics slow-growing liver cancer. Previous studies using murine models suggest that Echinococcus multilocularis (Em) metacestodes have developed mechanisms which impair the natural inflammatory host response. The aim of this study was to investigate in vitro the impact of Em vesicular fluid (VF) on monocytes, monocytes derived dendritic cells and lymphocytes from healthy blood donors.

Methods: First, assays were performed to investigate whether or not Em-VF influences monocyte-derived dendritic cell (MoDC) differentiation and maturation. Monocytes during differentiation and immature MoDCs were exposed to Em-VF. The effect of Em-VF was assessed using flow cytometry (CD86, CD83, CD80) and immune assays (IL-10 and TGFβ). Second, assays were performed to investigate the interaction between Em-VF, peripheral blood monocyte cells (PBMC) and Toll-like Receptor (TLR) agonists (LPS, PolyIC, R848 and CpG). PBMC were stimulated by each of the TLR agonists with and without Em-VF. The subsequent TGFβ production was assessed.

Results: Exposure to Em-VF had bearing on both differentiation and maturation of MoDC, but only partially. A decrease in the expression of co-stimulatory molecules was observed; however, levels of immune-regulatory cytokines were stable. PBMC exposed simultaneously to Em-VF and LPS induced a significant increase of TGFβ (p<0.05, Wilcoxon signed-rank test). Further experiments showed that TGFβ production was lymphocyte-dependent.

Conclusion: The assays performed confirmed that Em-VF influences the host immune response. However, only minor changes were observed when investigating the Em-VF impact on cells from healthy blood donors. Assays with TLR agonists suggested that co-stimulation with LPS reinforces the response of healthy blood donors exposed to Em-VF.

Keywords: Dendritic cells; Echinococcus multilocularis; Lymphocytes; Monocytes; TGFß; Vesicular fluid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / biosynthesis
  • B7-1 Antigen / biosynthesis
  • B7-2 Antigen / biosynthesis
  • CD83 Antigen
  • Cell Differentiation
  • Dendritic Cells / cytology*
  • Dendritic Cells / immunology
  • Echinococcosis
  • Echinococcosis, Hepatic / immunology*
  • Echinococcosis, Hepatic / parasitology
  • Echinococcus multilocularis / immunology*
  • Humans
  • Immunoglobulins / biosynthesis
  • Inflammation
  • Interleukin-10 / biosynthesis
  • Lipopolysaccharides
  • Lymphocyte Activation / immunology
  • Lymphocytes / immunology
  • Membrane Glycoproteins / biosynthesis
  • Monocytes / cytology*
  • Monocytes / immunology
  • Toll-Like Receptor 2 / immunology
  • Toll-Like Receptor 4 / immunology
  • Transforming Growth Factor beta / biosynthesis

Substances

  • Antigens, CD
  • B7-1 Antigen
  • B7-2 Antigen
  • CD86 protein, human
  • IL10 protein, human
  • Immunoglobulins
  • Lipopolysaccharides
  • Membrane Glycoproteins
  • TLR2 protein, human
  • TLR4 protein, human
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Transforming Growth Factor beta
  • Interleukin-10

Supplementary concepts

  • Alveolar echinococcosis