Role of Ca/CaN/NFAT Signaling in IL-4 Expression by Splenic Lymphocytes Exposed to Phthalate (2-ethylhexyl) Ester in Spleen Lymphocytes

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2014 Jan 15

PMID 24420859

Citations 4

Authors

Xiucong Pei

Zhiwen Duan

Mingyue Ma

Yuming Zhang

Li Guo

Affiliations

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Abstract

The aims of present study were to investigate the effect of phthalate (2-ethylhexyl) ester (DEHP) and mono-(2-ethylhexyl) phthalate (MEHP) on Th1/Th2 balance signaling for interleukin 4 (IL-4) expression in splenic lymphocytes, and contribution of MEHP to any hypothesized changes in vitro. Primary splenic lymphocytes were exposed to DEHP/MEHP. ELISA and Western blotting were used to detect proteins. Confocal-microscopy was used to examine nuclear translocation. Nuclear factor of activated T cells (NFAT) DNA binding activity was examined by electrophoretic mobility-shift assay. DEHP significantly increased IL-4 and interferon gamma (IFN-γ) level, and reduced Th1/Th2 ratio (reflected by IFN-γ/IL-4) with 5 μg/L Concanavalin A (ConA) treatment. While MEHP reduced Th1/Th2 ratio (represented by IFN-γ/IL-6). IL-4 mRNA was significantly increased by DEHP but not by MEHP after PMA and Ion treatment. DEHP significantly inhibited NFATp protein in cytosol and nucleus. DEHP augmented nuclear translocation of NFATc in transfected EL4 cells and NFAT DNA-binding activity. DEHP-mediated enhancement of calcium-dependent phosphatase calcineurin (CaN) protein, and NFAT and IL-4 expression were abrogated by calcium antagonist verapamil and CaN inhibitor tarcolimus. Ca(2+)/calmodulin antagonist chlorpromazine significantly suppressed IL-4 and CaN production with no NFAT mRNA change. Our study suggests that DEHP and MEHP impact Th1/Th2 balance by modulating different cytokines. DEHP-affected IL-4 expression through Ca/CaN/NFAT signaling pathway, but no effect was discovered for MEHP.

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References

Li X, Liang Y, Su Y, Deng H, Li X, Guo J . Adverse effects of di-(2-ethylhexyl) phthalate on Leydig cell regeneration in the adult rat testis. Toxicol Lett. 2012; 215(2):84-91. DOI: 10.1016/j.toxlet.2012.10.001. View

Gachet S, Ghysdael J . Calcineurin/NFAT signaling in lymphoid malignancies. Gen Physiol Biophys. 2010; 28 Spec No Focus:F47-54. View

Butala J, David R, Gans G, McKee R, Guo T, Peachee V . Phthalate treatment does not influence levels of IgE or Th2 cytokines in B6C3F1 mice. Toxicology. 2004; 201(1-3):77-85. DOI: 10.1016/j.tox.2004.04.004. View

Khoo M, Li J, Singh M, Yang Y, Kannankeril P, Wu Y . Death, cardiac dysfunction, and arrhythmias are increased by calmodulin kinase II in calcineurin cardiomyopathy. Circulation. 2006; 114(13):1352-9. DOI: 10.1161/CIRCULATIONAHA.106.644583. View

Casolaro V, Georas S, Song Z, Ono S . Biology and genetics of atopic disease. Curr Opin Immunol. 1996; 8(6):796-803. DOI: 10.1016/s0952-7915(96)80007-0. View

Luo C, Burgeon E, Rao A . Mechanisms of transactivation by nuclear factor of activated T cells-1. J Exp Med. 1996; 184(1):141-7. PMC: 2192690. DOI: 10.1084/jem.184.1.141. View

Bornehag C, Sundell J, Weschler C, Sigsgaard T, Lundgren B, Hasselgren M . The association between asthma and allergic symptoms in children and phthalates in house dust: a nested case-control study. Environ Health Perspect. 2004; 112(14):1393-7. PMC: 1247566. DOI: 10.1289/ehp.7187. View

Umetsu D . Revising the immunological theories of asthma and allergy. Lancet. 2005; 365(9454):98-100. DOI: 10.1016/S0140-6736(05)17714-9. View

Guo L, Urban J, Zhu J, Paul W . Elevating calcium in Th2 cells activates multiple pathways to induce IL-4 transcription and mRNA stabilization. J Immunol. 2008; 181(6):3984-93. PMC: 2744309. DOI: 10.4049/jimmunol.181.6.3984. View

10.

Robinson D, Hamid Q, Ying S, Tsicopoulos A, Barkans J, Bentley A . Predominant TH2-like bronchoalveolar T-lymphocyte population in atopic asthma. N Engl J Med. 1992; 326(5):298-304. DOI: 10.1056/NEJM199201303260504. View

11.

van der Pluijm I, Garinis G, Brandt R, Gorgels T, Wijnhoven S, Diderich K . Impaired genome maintenance suppresses the growth hormone--insulin-like growth factor 1 axis in mice with Cockayne syndrome. PLoS Biol. 2007; 5(1):e2. PMC: 1698505. DOI: 10.1371/journal.pbio.0050002. View

12.

Oie L, Hersoug L, Madsen J . Residential exposure to plasticizers and its possible role in the pathogenesis of asthma. Environ Health Perspect. 1997; 105(9):972-8. PMC: 1470354. DOI: 10.1289/ehp.97105972. View

13.

Glue C, Millner A, Bodtger U, Jinquan T, Poulsen L . In vitro effects of monophthalates on cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells from allergic and non-allergic donors. Toxicol In Vitro. 2002; 16(6):657-62. DOI: 10.1016/s0887-2333(02)00082-6. View

14.

Piccinni M, Maggi E, Romagnani S . Environmental factors favoring the allergen-specific Th2 response in allergic subjects. Ann N Y Acad Sci. 2001; 917:844-52. DOI: 10.1111/j.1749-6632.2000.tb05450.x. View

15.

Timmerman L, Healy J, Ho S, Chen L, Goodnow C, Crabtree G . Redundant expression but selective utilization of nuclear factor of activated T cells family members. J Immunol. 1997; 159(6):2735-40. View

16.

Palleschi S, Rossi B, Diana L, Silvestroni L . Di(2-ethylhexyl)phthalate stimulates Ca(2+) entry, chemotaxis and ROS production in human granulocytes. Toxicol Lett. 2009; 187(1):52-7. DOI: 10.1016/j.toxlet.2009.01.031. View

17.

Zarain-Herzberg A, Fragoso-Medina J, Estrada-Aviles R . Calcium-regulated transcriptional pathways in the normal and pathologic heart. IUBMB Life. 2011; 63(10):847-55. DOI: 10.1002/iub.545. View

18.

Davis A, Murphy C, Saraceni-Richards C, Rosenstein M, Wiegers T, Mattingly C . Comparative Toxicogenomics Database: a knowledgebase and discovery tool for chemical-gene-disease networks. Nucleic Acids Res. 2008; 37(Database issue):D786-92. PMC: 2686584. DOI: 10.1093/nar/gkn580. View

19.

Su E, Chang J, Cheng C, Sung T, Hsu W . Prediction of nuclear proteins using nuclear translocation signals proposed by probabilistic latent semantic indexing. BMC Bioinformatics. 2013; 13 Suppl 17:S13. PMC: 3521467. DOI: 10.1186/1471-2105-13-S17-S13. View

20.

Ranger A, Gerstenfeld L, Wang J, Kon T, Bae H, Gravallese E . The nuclear factor of activated T cells (NFAT) transcription factor NFATp (NFATc2) is a repressor of chondrogenesis. J Exp Med. 2000; 191(1):9-22. PMC: 2195796. DOI: 10.1084/jem.191.1.9. View