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Cell J. 2015;16(4):528-37. doi: 10.22074/cellj.2015.497. Epub 2015 Jan 13.

Effect of Interleukin-29 on Interferon-α Secretion by Peripheral Blood Mononuclear Cells.

Cell journal

Chi Hyun Cho, Soo Young Yoon, Chang Kyu Lee, Chae Seung Lim, Yunjung Cho

Affiliations

  1. Department of Laboratory Medicine, College of Medicine, Korea University, Seoul, Korea.

PMID: 25685743 PMCID: PMC4297491 DOI: 10.22074/cellj.2015.497

Abstract

OBJECTIVE: The effect of interleukin (IL)-29, a new therapeutic agent similar to type I interferons (IFNs), on IFN-α secretion of human plasmacytoid dendritic cells (pDCs) has not been studied. Therefore, in this study, we aimed to clarify the effect of IL-29 on IFN-α secretion of pDCs using human peripheral blood mononuclear cells (PBMCs) in the presence of cytosine-phosphate-guanosinemotif-containing oligodeoxy nucleotides (CpG).

MATERIALS AND METHODS: In this experimental and prospective study, PBMCs were ob- tained from 11 healthy volunteers and divided into four culture conditions: I. control, II. CpG treatment, III. IL-29 treatment and IV. CpG plus IL-29 treatment. The amount of IFN-α secretion was measured from each culture supernatant by flow cytometry using the flowcytomix apparatus (eBioscience, Vienna, Austria). Fractional IFN-α production of the cultured PBMCs was measured by intracellular staining using the cytomics FC 500 system (Beckman Coulter, Brea, CA, USA) with CXP Software.

RESULTS: The mean ± standard deviation (SD) of supernatant IFN-α secretion per pDC/μL was 5.7 ± 9.3 pg/mL/count/µL for condition I, 1071.5 ± 1026.6 pg/mL/count/µL for condition II, 14.1 ± 21.1 pg/mL/count/µL for condition III, and 1913.9 ± 1525.9 pg/mL/count/µL for condition IV. There were statistically significant differences between conditions I and II as well as betweenconditions II and IV. Intracellular IFN-α production was only detectable in the pDC fraction from one culture; the production amount was similar between the cells treated with CpG and those treated with CpG plus IL-29. Natural killer (NK) cell production of IFN-α was observed in two out of three cultures and one culture showed IFN- α production in the monocyte fraction.

CONCLUSION: IL-29 alone did not show any effect on IFN-α secretion of PBMCs. However, the addition of CpG along with IL-29 enhanced IFN-α secretion of PBMCs. Given that pDCs are the major secretors of IFN-α in peripheral blood, this result has suggested the possibility that IL-29 has an enhancing effect in human pDC IFN-α secretion. Although the supernatant IFN-α secretion was not directly correlated with pDCs's intracellular IFN-α production in this study, prolonged incubation of pDC and other PB subsets with CpG or IL-29 for over 4 hours could be applied in future studies. These studies would help to elucidate the mechanism of action of IL-29 in human pDCs associated with viral infections.

Keywords: CpG; IFN-α; IL-29; Peripheral Blood; Plasmacytoid Dendritic Cells

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