Skip to main content
SpringerLink
Log in

Suppressive effect of CTB glycoprotein (75 kDa) on IL-4 expression in primary-cultured lymphocytes treated with di(2-ethylhexyl) phthalate

  • ORIGINAL ARTICLE
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

The purpose of this study is to determine the inhibitory effect of a glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein, 75 kDa) on di(2-ethylhexyl) phthalate (DEHP)-induced differentiation of T helper (Th) type 2 cells in T lymphocytes separated from mice. This experiment evaluated the activities of protein kinase C (PKC), mitogen-activated protein kinase (MAPK), transcription factors [signal transducer and activator of transcription (STAT)-6 and GATA-binding protein 3 (GATA3)], and Th2 cell-related cytokine [interleukin-4 (IL-4)] using immunoblotting and reverse transcription-polymerase chain reaction. Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits the translocation of PKC from cytosol to membrane and the phosphorylation of p44/42 MAPK in primary cultured T lymphocytes. We also found that the CTB glycoprotein (100 μg/ml) has suppressive effects on transcriptional activation of STAT6, GATA3, and on the expression level of IL-4 in DEHP-treated T lymphocytes. The phosphorylation of STAT6 and GATA3 were collectively blocked by treatment with PKC inhibitor (staurosporine) as well as p44/42 MAPK inhibitor (PD 98059), respectively. The results from these experiments indicated that the CTB glycoprotein inhibits IL-4 expression, not IL-10 expression, on the stage of Th2 cell differentiation induced by DEHP in T lymphocytes. Hence, we speculate that the CTB glycoprotein has a strong inhibitory ability for expressions of allergy-related cytokines indirectly caused by DEHP in Th2 cell differentiation of the primary cultured mouse T lymphocytes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Baier G (2007) PKC isotype functions in T lymphocytes. Ernst Schering Found Symp Proc 3:29–41

    Article  PubMed  Google Scholar 

  • Chalubinski M, Kowalski ML (2006) Endocrine disrupters—potential modulators of the immune system and allergic response. Allergy 61:1326–1335

    Article  PubMed  CAS  Google Scholar 

  • Christodoulopoulos P, Cameron L, Nakamura Y, Lemiere C, Muro S, Dugas M, Boulet LP, Laviolette M, Olivenstein R, Hamid Q (2001) Th2 cytokine-associated transcription factors in atopic and nonatopic asthma: evidence for differential signal transducer and activator of transcription 6 expression. J Allergy Clin Immunol 107:586–591

    Article  PubMed  CAS  Google Scholar 

  • Ema M, Miyawaki E (2001) Effects of monobutyl phthalate on reproductive function in pregnant and pseudopregnant rats. Reprod Toxicol 15:261–267

    Article  PubMed  CAS  Google Scholar 

  • Gourlay T, Samartzis I, Stefanou D, Taylor K (2003) Inflammatory response of rat and human neutrophils exposed to di-(2-ethyl-hexyl)-phthalate-plasticized polyvinyl chloride. Artif Organs 27:256–260

    Article  PubMed  CAS  Google Scholar 

  • Groux H, O’Garra A, Bigler M, Rouleau M, Antonenko S, de Vries JE, Roncarolo MG (1997) A CD4 + T-cell subset inhibits antigenspecific T-cell responses and prevents colitis. Nature 389:737–742

    Article  PubMed  CAS  Google Scholar 

  • Herrick CA, MacLeod H, Glusac E, Tigelaar RE, Bottomly K (2000) Th2 responses induced by epicutaneous or inhalational protein exposure are differentially dependent on IL-4. J Clin Invest 105:765–775

    Article  PubMed  CAS  Google Scholar 

  • Joo HY, Lim KT (2009) Glycoprotein isolated from Cudrania tricuspidata Bureau inhibits iNO and COX-2 expression through modulation of NF-κB in LPS-stimulated RAW 264.7 cells. Environ Toxicol Pharmacol 27:247–252

    Article  CAS  Google Scholar 

  • Kaplan MH, Schnidler U, Smiley ST, Grusby MJ (1996) Stat6 is required for mediating responses to IL-4 and for the development of Th2 cells. Immunity 4:313–319

    Article  PubMed  CAS  Google Scholar 

  • Kato Y, Negishi T, Furusako S, Mizuguchi K, Mochizuki H (2003) An orally active Th1/Th2 balance modulator, M50367, suppresses Th2 differentiation of naive Th cell in vitro. Cell Immunol 224:29–37

    Article  PubMed  CAS  Google Scholar 

  • Larsen ST, Lund RM, Nielsen GD, Thygesen P, Poulsen OM (2002) Adjuvant effect of di-n-butyl-, di-n-octyl-, di-iso-nonyl- and di-iso-decyl phthalate in a subcutaneous injection model using BALB/c mice. Pharmacol Toxicol 91:264–272

    Article  PubMed  CAS  Google Scholar 

  • Larsen ST, Hansen JS, Hansen EW, Clausen PA, Nielsen GD (2007) Airway inflammation and adjuvant effect after repeated airborne exposures to di-(2-ethylhexyl) phthalate and ovalbumin in BALB/c mice. Toxicology 235:119–129

    Article  PubMed  CAS  Google Scholar 

  • Lee MH, Park J, Chung SW, Kang BY, Kim SH, Kim TS (2004) Enhancement of interleukin-4 production in activated CD4 + T cells by diphthalate plasticizers via increased NFAT binding activity. Int Arch Allergy Immunol 134:213–222

    Article  PubMed  CAS  Google Scholar 

  • Li XM, Huang CK, Zhang TF, Teper AA, Srivastava K, Schofield BH, Sampson HA (2000) The Chinese herbal medicine formula MSSN-002 suppresses allergic airway hyperreactivity and modulates Th1/Th2 responses in a murine model of allergic asthma. J Allergy Clin Immunol 106:660–668

    Article  PubMed  CAS  Google Scholar 

  • Liblau RS, Singer SM, McDevitt HO (1995) Th1 and Th2 CD4 + T cells in the pathogenesis of organ-specific autoimmune diseases. Immunol Today 16:34–38

    Article  PubMed  CAS  Google Scholar 

  • Lovekamp-Swan T, Davis BJ (2003) Mechanisms of phthalate ester toxicity in the female reproductive system. Environ Health Perspect 111:139–145

    PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  PubMed  CAS  Google Scholar 

  • Murphy KM, Reiner SL (2002) The lineage decisions of helper T cells. Nat Rev Immunol 2:933–944

    Article  PubMed  CAS  Google Scholar 

  • Murphy KM, Ouyang W, Farrar JD, Yang J, Ranganath S, Asnagli H, Afkarian M, Murphy TL (2000) Signaling and transcription in T helper development. Annu Rev Immunol 18:451–494

    Article  PubMed  CAS  Google Scholar 

  • Nakamura Y, Christodoulopoulos P, Cameron L, Wright E, Lavigne F, Toda M, Muro S, Ray A, Eidelman DH, Minshall E, Hamid Q (2000) Upregulation of the transcription factor GATA-3 in upper airway mucosa after in vivo and in vitro allergen challenge. J Allergy Clin Immunol 105:1146–1152

    Article  PubMed  CAS  Google Scholar 

  • Neville DM Jr, Glossmann H (1974) Molecular weight determination of membrane protein and glycoprotein subunits by discontinuous gel electrophoresis in dodecyl sulfate. Methods Enzymol 32:92–102

    Article  PubMed  CAS  Google Scholar 

  • O’Garra A, Murphy K (1994) Role of cytokines in determining T-lymphocyte function. Curr Opin Immunol 6:458–466

    Article  PubMed  Google Scholar 

  • Oh PS, Lim KT (2006) Glycoprotein (90 kDa) isolated from Opuntia ficus-indica var. saboten MAKINO lowers plasma lipid level through scavenging of intracellular radicals in Triton WR-1339-induced mice. Biol Pharm Bull 29:1391–1396

    Article  PubMed  CAS  Google Scholar 

  • Oh PS, Lim KT (2008) Protective activity of 30 kDa phytoglycoprotein from glucose/glucose oxidase-induced cell death in primary cultured mouse thymocytes. Environ Toxicol Pharmacol 25:114–120

    Article  CAS  Google Scholar 

  • Oh PS, Lee SJ, Lim KT (2007) Glycoprotein isolated from Rhus verniciflua Stokes inhibits inflammation-related protein and nitric oxide production in LPS-stimulated RAW 264.7 cells. Biol Pharm Bull 30:111–116

    Article  PubMed  CAS  Google Scholar 

  • Ooi VE, Liu F (2000) Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Curr Med Chem 7:715–729

    PubMed  CAS  Google Scholar 

  • Park KH, Park YD, Han JM, Im KR, Lee BW, Jeong IY, Jeong TS, Lee WS (2006) Anti-atherosclerotic and anti-inflammatory activities of catecholic xanthones and flavonoids isolated from Cudrania tricuspidata. Bioorg Med Chem Lett 16:5580–5583

    Article  PubMed  CAS  Google Scholar 

  • Parronchi P, De Carli M, Manetti R, Simonelli C, Sampognaro S, Piccinni MP, Macchia D, Maggi E, Del Prete G, Romagnani S (1992) IL-4 and IFN (alpha and gamma) exert opposite regulatory effects on the development of cutolytic potential by Th1 or Th2 human T cell clones. J Immunol 149:2977–2983

    PubMed  CAS  Google Scholar 

  • Patton WF, Dhanak MR, Jacobson BS (1989) Differential partitioning of plasma membrane proteins into the triton X-100-insoluble cytoskeleton fraction during concanavalin A-induced receptor redistribution. J Cell Sci 92:85–91

    PubMed  CAS  Google Scholar 

  • Pierce A, Heyworth CM, Nicholls SE, Spooncer E, Dexter TM, Lord JM, Owen-Lynch PJ, Wark G, Whetton AD (1998) An activated protein kinase C alpha gives a differentiation signal for hematopoietic progenitor cells and mimicks macrophage colony-stimulating factorstimulated signaling events. J Cell Biol 140:1511–1518

    Article  PubMed  CAS  Google Scholar 

  • Schettler T (2006) Human exposure to phthalates via consumer products. Int J Androl 29:134–139

    Article  PubMed  CAS  Google Scholar 

  • Shimoda K, Deursen J, Sangster MY, Sarawar S, Carson RT, Tripp RA, Chu C, Quelle FW, Nosaka T, Vignali DAA, Doherty PC, Grosveld G, Paul WE, Ihle JN (1996) Lack of IL-4-induced Th2 response and IgE class switching in mice with disrupted Stat6 gene. Nature 380:630–633

    Article  PubMed  CAS  Google Scholar 

  • Spellberg B, Edwards JE (2001) Type 1/type 2 immunity in infectious diseases. Clin Infect Dis 32:76–102

    Article  PubMed  CAS  Google Scholar 

  • Verhagen J, Taylor A, Blaser K, Akdis M, Akdis CA (2005) T regulatory cells in allergen-specific immunotherapy. Int Rev Immunol 24:533–548

    Article  PubMed  CAS  Google Scholar 

  • Yamashita M, Kimura M, Kubo M, Shimizu C, Tada T, Perlmutter RM, Nakayama T (1999) T cell antigen receptor-mediated activation of the Ras/mitogen-activated protein kinase pathway controls interleukin 4 receptor function and type-2 helper T cell differentiation. Proc Natl Acad Sci U S A 96:1024–1029

    Article  PubMed  CAS  Google Scholar 

  • Yamashita M, Shinnakasu R, Asou H, Kimura M, Hasegawa A, Hashimoto K, Hatano N, Ogata M, Nakayama T (2005) Ras-ERK MAPK cascade regulates GATA3 stability and Th2 differentiation through ubiquitin-proteasome pathway. J Biol Chem 280:29409–29419

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Korea Research Foundation Grant funded by the Korean government (MOEHRD, Basic Research Promotion Fund; KRF-2008-521-C00167).

Author information

Authors and Affiliations

  1. Molecular Biochemistry Laboratory, Biotechnology Research Institute & Center for the Control of Animal Hazards Using Biotechnology (BK21), Chonnam National University, 300 Yongbong-Dong, Gwang-ju, 500-757, South Korea

    Phil-Sun Oh

  2. Molecular Biochemistry Laboratory, Biotechnology Research Institute, Chonnam National University, 300 Yongbong-Dong, Gwang-ju, 500-757, South Korea

    Kye-Taek Lim

Authors
  1. Phil-Sun Oh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kye-Taek Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kye-Taek Lim.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Oh, PS., Lim, KT. Suppressive effect of CTB glycoprotein (75 kDa) on IL-4 expression in primary-cultured lymphocytes treated with di(2-ethylhexyl) phthalate. Naunyn-Schmied Arch Pharmacol 380, 115–124 (2009). https://doi.org/10.1007/s00210-009-0423-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00210-009-0423-y

Keywords

Search

Navigation