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Ionizing radiation affects the expression of Toll-like receptors 2 and 4 in human monocytic cells through c-Jun N-terminal kinase activation.

Yoshino H, Chiba K, Saitoh T, Kashiwakura I - J. Radiat. Res. (2014)

Bottom Line: Pattern recognition receptors recognize pathogen-associated molecular patterns.X-irradiation caused increased expression of these TLRs in THP1 and decreased expression in macrophage-like cells.However, although TNF-α expression was not affected by X-irradiation in macrophage-like cells, the expression of LPS-inducible interferon-β was lower following X-irradiation of macrophage-like cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiological Life Sciences, Division of Medical Life Sciences, Hirosaki University Graduate School of Health Sciences, 66-1 Hon-cho, Hirosaki, Aomori 036-8564, Japan.

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Cell morphology and viable cell numbers of X-irradiated THP1 monocytes and macrophage-like cells. (A) Morphology of THP1 monocytes (left) and macrophage-like cells (right). (B) THP1 monocytes were exposed to X-irradiation and were cultured for 24–72 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. (C) Macrophage-like cells were exposed to X-irradiation and were cultured for 24 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. Data are presented as the mean ± SD of four independent experiments; single and double asterisks indicate P < 0.05 and P < 0.01 compared with non-irradiated control, respectively; n.s. = not significant.
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RRU040F1: Cell morphology and viable cell numbers of X-irradiated THP1 monocytes and macrophage-like cells. (A) Morphology of THP1 monocytes (left) and macrophage-like cells (right). (B) THP1 monocytes were exposed to X-irradiation and were cultured for 24–72 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. (C) Macrophage-like cells were exposed to X-irradiation and were cultured for 24 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. Data are presented as the mean ± SD of four independent experiments; single and double asterisks indicate P < 0.05 and P < 0.01 compared with non-irradiated control, respectively; n.s. = not significant.

Mentions: THP1-derived macrophages (macrophage-like cells) were prepared as previously described [14]. Briefly, THP1 monocytes (2.0 × 105 cells/ml) were plated in 60 mm dishes (IWAKI, Tokyo, Japan) with 4 ml of medium containing 100 ng/ml PMA, and were cultured for 48 h. Differentiation to macrophage-like cells was confirmed by observing morphological changes under a microscope, as shown in Fig. 1A. After 48-h culture, the medium containing PMA was replaced with fresh medium not containing PMA, and macrophage-like cells were then used in experiments. Adherent macrophage-like cells were harvested by trypsinization with 0.1% trypsin/ethylenediaminetetraacetate (Gibco). Viable cell numbers were estimated using trypan blue dye exclusion assays.Fig. 1.


Ionizing radiation affects the expression of Toll-like receptors 2 and 4 in human monocytic cells through c-Jun N-terminal kinase activation.

Yoshino H, Chiba K, Saitoh T, Kashiwakura I - J. Radiat. Res. (2014)

Cell morphology and viable cell numbers of X-irradiated THP1 monocytes and macrophage-like cells. (A) Morphology of THP1 monocytes (left) and macrophage-like cells (right). (B) THP1 monocytes were exposed to X-irradiation and were cultured for 24–72 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. (C) Macrophage-like cells were exposed to X-irradiation and were cultured for 24 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. Data are presented as the mean ± SD of four independent experiments; single and double asterisks indicate P < 0.05 and P < 0.01 compared with non-irradiated control, respectively; n.s. = not significant.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4202298&req=5

RRU040F1: Cell morphology and viable cell numbers of X-irradiated THP1 monocytes and macrophage-like cells. (A) Morphology of THP1 monocytes (left) and macrophage-like cells (right). (B) THP1 monocytes were exposed to X-irradiation and were cultured for 24–72 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. (C) Macrophage-like cells were exposed to X-irradiation and were cultured for 24 h. Cells were harvested and viable cell numbers were estimated using the trypan blue dye exclusion method. Data are presented as the mean ± SD of four independent experiments; single and double asterisks indicate P < 0.05 and P < 0.01 compared with non-irradiated control, respectively; n.s. = not significant.
Mentions: THP1-derived macrophages (macrophage-like cells) were prepared as previously described [14]. Briefly, THP1 monocytes (2.0 × 105 cells/ml) were plated in 60 mm dishes (IWAKI, Tokyo, Japan) with 4 ml of medium containing 100 ng/ml PMA, and were cultured for 48 h. Differentiation to macrophage-like cells was confirmed by observing morphological changes under a microscope, as shown in Fig. 1A. After 48-h culture, the medium containing PMA was replaced with fresh medium not containing PMA, and macrophage-like cells were then used in experiments. Adherent macrophage-like cells were harvested by trypsinization with 0.1% trypsin/ethylenediaminetetraacetate (Gibco). Viable cell numbers were estimated using trypan blue dye exclusion assays.Fig. 1.

Bottom Line: Pattern recognition receptors recognize pathogen-associated molecular patterns.X-irradiation caused increased expression of these TLRs in THP1 and decreased expression in macrophage-like cells.However, although TNF-α expression was not affected by X-irradiation in macrophage-like cells, the expression of LPS-inducible interferon-β was lower following X-irradiation of macrophage-like cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiological Life Sciences, Division of Medical Life Sciences, Hirosaki University Graduate School of Health Sciences, 66-1 Hon-cho, Hirosaki, Aomori 036-8564, Japan.

Show MeSH
Related in: MedlinePlus