Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Jun;156(3):405-12.
doi: 10.1111/j.1365-2249.2009.03911.x.

Combination use of immune complexes and a Ca2(+) channel blocker azelnidipine enhances interleukin-12 p40 secretion without T helper type 17 cytokine secretion in human monocyte-derived dendritic cells

T Abe  1 , I Fuse, M Narita, M Takahashi, Y Aizawa
Affiliations

Combination use of immune complexes and a Ca2(+) channel blocker azelnidipine enhances interleukin-12 p40 secretion without T helper type 17 cytokine secretion in human monocyte-derived dendritic cells

T Abe et al. Clin Exp Immunol. 2009 Jun.

Abstract

Immune complexes (ICs) improve the capacity of priming specific CD8(+) cytotoxic T cell responses of dendritic cells (DCs). ICs induce phosphorylation of mitogen-activated protein kinases (MAPK) and calcium influx, although the precise regulating mechanism still remains unclear. In the present study, we investigated the effect of a Ca2(+) channel blocker on the phosphorylation of p38 MAPK and extracellular signal-regulated kinase (ERK) in immature monocyte-derived DCs stimulated with lipopolysaccharide (LPS) or LPS-ICs, and the production of interleukin (IL)-12 family members (p40, p70, IL-23), T helper type 17 (Th17) cytokines (IL-6 and IL-23), tumour necrosis factor (TNF)-alpha and IL-10 were also investigated. In comparison with LPS stimulation, LPS-ICs stimulation enhanced p38 MAPK phosphorylation significantly, which was associated with an increase in IL-12 p40 monomer/homodimer secretion. LPS-ICs also enhanced TNF-alpha and IL-6 secretion, but suppressed IL-23 secretion. The use of azelnidipine (Aze), a long-acting L-type Ca2(+) channel blocker with a high lipid solubility, suppressed p38 MAPK phosphorylation stimulated with LPS or LPS-ICs, but surprisingly enhanced IL-12 p40 monomer/homodimer secretion stimulated with LPS-ICs. This IL-12 p40 secretion-enhancing effect was not accompanied by IL-10 or IL-23 production, but was associated with ERK phosphorylation. The use of Aze did not affect IL-12 p70 production. These results suggest that the use of Aze enhances ICs-mediated IL-12 p40 secretion without additional IL-23 secretion. Therefore, the use of Aze and ICs could be a new therapeutic approach to immunomolecular therapy, as it does not cause Th17 differentiation which induces autoimmunity or reduces anti-tumour immunity.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Interleukin (IL)-12 p40 monomer and/or homodimer and p70 production from immature monocyte-derived dendritic cells (iMDDCs) stimulated with immunoglobulin (Ig), lipopolysaccharide (LPS) or LPS-immune complexes (ICs) in the presence or absence of azelnidipine (Aze). iMDDCs were pretreated with vehicle alone or Aze (1 µM) for 1 h and stimulated with vehicle alone, Ig, LPS or LPS-ICs for 12 h at 37°C. IL-12 p40 monomer and/or homodimer (a) and IL-12 p70 (b) production were measured in the supernatants of DCs with each enzyme-linked immunosorbent assay kit. Vertical bars represent mean ± standard error of the mean (a: n = 4, b: n = 3).
Fig. 2
Fig. 2
Effect of azelnidipine (Aze) on p38 mitogen-activated protein kinases (MAPK) phosphorylation in immature monocyte-derived dendritic cells (iMDDCs) stimulated with immunoglobulin (Ig), lipopolysaccharide (LPS) or LPS-immune complexes (ICs). iMDDCs were pretreated with vehicle alone or Aze (1 µM) for 1 h, and stimulated with Ig, LPS or LPS-ICs at the indicated time-intervals at 37°C, and whole lysates were prepared. Samples were analysed by Western blot with antibodies against the phosphorylated form of p38 MAPK, after which the membranes were reprobed with antibodies specific to total p38 MAPK. Representative results of three experiments are shown. The densitometric data from three independent experiments are expressed as mean phosphorylation ratio ± standard error of the mean. The ratio of phosphorylated protein to total protein band intensity was calculated.
Fig. 3
Fig. 3
Effect of azelnidipine (Aze) on extracellular signal-regulated kinase (ERK) phosphorylation in immature monocyte-derived dendritic cells (iMDDCs) stimulated with immunoglobulin (Ig), lipopolysaccharide (LPS) or LPS-immune complexes (ICs). iMDDCs were pretreated with vehicle alone or Aze (1 µM) for 1 h, and stimulated with Ig, LPS or LPS-ICs at the indicated time-intervals at 37°C, and whole lysates were prepared. Samples were analysed by Western blot with antibodies against the phosphorylated form of ERK, after which the membranes were reprobed with antibodies specific to total ERK. Representative results of three experiments are shown. The densitometric data from three independent experiments are expressed as mean phosphorylation ratio ± standard error of the mean. The ratio of phosphorylated protein to total protein band intensity was calculated.
Fig. 4
Fig. 4
Effect of PD98059 or SB203580 on interleukin (IL)-12 p40 monomer and/or homodimer secretion from immature monocyte-derived dendritic cells (iMDDCs) stimulated with lipopolysaccharide-immune complexes (LPS-ICs). iMDDCs were pretreated with vehicle alone, PD98059 (40 µM) or SB203580 (50 µM) for 1 h, and then stimulated with LPS-ICs for 12 h at 37°C. IL-12 p40 monomer and/or homodimer production was measured in the supernatants of DCs with IL-12 p40 enzyme-linked immunosorbent assay kit. Vertical bars represent mean ± standard error of the mean (n = 3).
Fig. 5
Fig. 5
Interleukin (IL)-23 secretion from immature monocyte-derived dendritic cells (iMDDCs) stimulated with immunoglobulin (Ig), lipopolysaccharide (LPS) or LPS-immune complexes (ICs) in the presence or absence of azelnidipine (Aze). iMDDCs were pretreated with vehicle alone or Aze (1 µM) for 1 h, and then stimulated with Ig, LPS or LPS-ICs for 12 h at 37°C. IL-23 production was measured in the supernatants of DCs with each enzyme-linked immunosorbent assay kit. Vertical bars represent mean ± standard error of the mean (n = 3).
Fig. 6
Fig. 6
Interleukin (IL)-10, tumour necrosis factor (TNF)-α and IL-6 secretion from immature monocyte-derived dendritic cells (iMDDCs) stimulated with immunoglobulin (Ig), lipopolysaccharide (LPS) or LPS-immune complexes (ICs) in the presence or absence of azelnidipine (Aze). iMDDCs were pretreated with vehicle alone or Aze (1 µM) for 1 h, and then stimulated with Ig, LPS or LPS-ICs for 12 h at 37°C. IL-10 (a), TNF-α (b) and IL-6 (c) production were measured in the supernatants of DCs with each enzyme-linked immunosorbent assay kit. Vertical bars represent mean ± standard error of the mean (n = 3).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Banchereau J, Steinman RM. Dendritic cells and the control of immunity. Nature. 1998;392:245–52. - PubMed
    1. Macatonia SE, Hosken NA, Litton M, et al. Dendritic cells produce IL-12 and direct the development of Th1 cells from naive CD4+ T cells. J Immunol. 1995;154:5071–9. - PubMed
    1. Trinchieri G, Scott P. Interleukin-12: a proinflammatory cytokine with immunoregulatory functions. Res Immunol. 1995;146:423–31. - PubMed
    1. Schuurhuis DH, Laban S, Toes RE, et al. Immature dendritic cells acquire CD8(+) cytotoxic T lymphocyte priming capacity upon activation by T helper cell-independent or -dependent stimuli. J Exp Med. 2000;192:145–50. - PMC - PubMed
    1. Sallusto F, Cella M, Danieli C, Lanzavecchia A. Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J Exp Med. 1995;182:389–400. - PMC - PubMed

MeSH terms