Baselet, B.

Benotmane, R.

Dettmering, T.

Durante, M.

Erbeldinger, N.

Fournier, C.

Liebig, M.

Lowe, D.

Raj, K.

Tapio, S.

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Cytokine release and adhesion of immune cells to cardiac endothelial cells ∗N. Erbeldinger1,5, M. Liebig1,5, T. Dettmering1, D. Lowe2, B. Baselet3, R. Benotmane3, S. Tapio4, K.Raj2, M. Durante1,5, and C. Fournier11GSI, Darmstadt, Germany; 2Public Health England, Oxford, UK; 3SCK-CEN, Belgium;4HelmholtzZentrumMu¨nchen, Germany; 5TU Darmstadt, Darmstadt, GermanyIntroductionThe cardiovascular system is nowadays well establishedas an organ of risk for accidental and therapeutic irradia-tion. Exposure of the cardiovascular system even with lowdoses (up to 2 Gy) can lead to an increased risk for the de-velopment of cardiovascular diseases (CVD) [1,2]. The un-derlying mechanism of developing CVD is putatively basedon changed cytokine release of vascular cells (endothelialcells and smooth muscle cells), although their interactivepattern is still not understood [3]. In order to investigatethe effect of low dosed X-irradiation on the first steps ofCVD, we measured cytokine release and adhesion of im-mune cells after irradiation of cardiac endothelial cells,mimicking the inner layer of an in vivo blood vessel. Fol-lowing we discuss the release of transforming growth factorβ1 (TGF-β1), a cytokine which can have both profibroticand anti-inflammatory effects, and its relevance on the ad-hesion of peripheral lymphocytes (PBL) to an endothelialmonolayer.Material and MethodsHuman coronary artery endothelial cells (HCAEC) wereimmortalized by est2 expression. For analysis of cytokinerelease, cells were cultured for 3 days prior irradiation. Af-ter irradiation with 0.05, 0.1, 0.5 and 2 Gy X-rays, mediumsupernatants were collected after 4 h, 24 h, 1 week and 2weeks and frozen at −80◦C. Medium was replaced afterirradiation/24 h before each time point. TGF-β1 concen-trations were quantified using ELISA (eBioscience). Forfunctional adhesion assays, FITC-tagged PBL were puri-fied from donor blood and incubated with an EC monolayer4 h/24 h after irradiation. The adhered PBL were micro-scopically quantified. Data were analyzed using a custompipeline written in the R language.Results and DiscussionIn Figure 1A the release of TGF-β1 after irradiation ofan EC monolayer is shown 4 h after irradiation. In compar-ison to the unirradiated control, there is a trend towards anincreased release after low doses (0.05 to 0.5 Gy), but nochange after high doses (2 Gy). In comparison to those re-sults, Figure 1B illustrates the relative adhesion of PBL toan EC monolayer. After irradiation with low doses (0.05 −∗The research leading to these results has received funding fromthe Euratom Seventh Framework Programme under grant agreement no.295823 (”PROCARDIO”).0.1 Gy), there is no change in adhesion of PBL detectable,whereas for high doses (0.5 − 2 Gy) increased adhesiontakes place.Figure 1: TGF-β1 release and adhesion of PBL 4 h afterirradiation of an EC monolayer. N=2, n=6.Cytokine release is well described to mediate attractionof immune cells to damaged tissue, i.e. after irradiation[4]. TGF-β1 has pleiotropic properties, since it is a knownanti-inflammatory and profibrotic cytokine. The presentedresults 4 h after irradiation show correlation between nochanges in relative adhesion of immune cells and increasedTGF-β1 release for low doses (0.05 − 0.1 Gy). After irra-diation with the highest dose X-ray (2 Gy), no changes inTGF-β1 release can be detected in parallel to an increasedrelative adhesion. This implies dependence between ad-hesion of immune cells and TGF-β1 release, but cannotexplain the increased TGF-β1 release and in parallel in-creased adhesion after 0.5 Gy. This points toward an in-fluence of TGF-β1 on the adhesion of immune cells, butshows no exclusive interference between both mechanisms.This is in good agreement of the current dogma, that stimu-lation of the immune system is a complex process, orches-trated by multiple cytokines and other factors.References[1] Boerma and Hauer-Jensen. Cardiol Res Pract. 2010 Oct4;2011. pii: 858262.[2] Adams et al. Semin Radiat Oncol. 2003 Jul; 13(3):34656.[3] Schultz-Hector et al. Int J Radiat Oncol Biol Phys 2007 Jan1; 67(1):108.[4] Libby et al. Nature. 2011 May 19;473(7347):317-25. doi:10.1038/nature10146.APPA-HEALTH-35 GSI SCIENTIFIC REPORT 2014328 DOI:10.15120/GR-2015-1-APPA-HEALTH-35

Cytokine release and adhesion of immune cells to cardiac endothelial cells

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Cytokine release and adhesion of immune cells to cardiac endothelial cells

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