Skip to main content
SpringerLink
Log in

Lack of direct immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on human peripheral blood lymphocyte subsets in vitro

  • Original Investigations
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

A wide variety of immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in experimental animals has been documented. In contrast, the impact of dioxin on the human immune system remains controversial, although adverse health effects have been reported in humans after occupational or accidental exposure to dioxin. Recently, Neubert et al. (1991) found that a dose-dependent decrease of peripheral blood lymphocyte (PBL) subpopulations in humans and non-human primates, including helper-inducer/memory cells (CD4+CD29+) and B cells (CD20+) occurred in pokeweed mitogen (PWM) stimulated cultures at concentrations as low as 10−12–10−14 M TCDD. Therefore, the direct effects of dioxin on human PBL subpopulations have been studied, in order to determine their usefulness as sensitive biomarkers for human dioxin exposure. Lymphocyte cultures from healthy individuals were treated with 10−7 M–10−14 M TCDD in the absence and presence of stimulation with pokeweed mitogen (PWM) or anti-CD3 monoclonal antibody (moAb; OKT3) for 3 days. Cytochrome P450 (CYP1A1) enzyme induction, one of the best studied direct biological effects of TCDD on numerous cell types, was assayed in parallel by ethoxyresorufin-O-deethylase (EROD) activity. Percentages of the different lymphocytes subsets, including CD2 (T cells); CD4; CD45 RA (suppressor-inducer/virgin T cells); CD4 CD29; CD8; CD19 (B cells) as well as interleukin 2 (IL-2) receptor (CD25) and class II antigen (HLA-DR) expression, were anlayzed by flow cytometry. DNA synthesis was determined by3H-thymidine uptake after 3 days of culture. In the present study, all stimulated lymphocyte cultures showed a dose-dependent significant increase of CYP1A1 activity at dioxin concentrations of 10−7 and 10−9 M. No enzyme activity could be detected at lower concentrations of TCDD. On the other hand, neither alteration in surface marker distribution nor suppression of lymphocyte proliferation could be demonstrated in mitogen-activated cells following any concentration of TCDD treatment. These data suggest that the inducibility of CYP1A1 enzyme activity is not correlated with direct immunotoxic effects in vitro in human PBL. In contrast to a previous report by Neubert et al. (1991), lymphoproliferation and phenotypes of human PBL are resistant to dioxin exposure in vitro and therefore appeared not to be useful as sensitive biomarkers in human exposure studies.

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

Similar content being viewed by others

References

  • Bekesi JG, Holland JF, Anderson HA et al. (1978) Lymphocyte function of Michigan dairy farmers exposed to polybrominated biphenyls. Science 199: 1207–1209

    Article  PubMed  CAS  Google Scholar 

  • Birnbaum LS (1994) The mechanism of dioxin toxicity: relationship to risk assessment. Environ Health Perspect (in press)

  • Boyum A (1968) Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Lab Invest 21: 77–89

    Article  CAS  Google Scholar 

  • Chang K-J, Hsieh K-H, Lee T-P, Tang S-Y, Tung T-C (1981) Immunologic evaluation of patients with polychlorinated biphenyl poisoning: determination of lymphocyte subpopulations. Toxicol Appl Pharmacol 61: 58–63

    Article  PubMed  CAS  Google Scholar 

  • Chang K-J, Hsieh K-H, Tang S-Y, Tung T-C, Lee T-P (1982) Immunologic evaluation of patients with polychlorinated biphenyl poisoning: evaluation of delayed-type skin hypersensitivity response and its relation to clinical studies. J Toxicol Environ Health 9: 217–223

    Article  PubMed  CAS  Google Scholar 

  • Clark G, Poland A, Glover E, McCoy Z, Lucier G (1992a) Expression of the Ah receptor in human peripheral blood lymphocytes. In: Organohalogen compounds, volume 10: Toxicology, epidemiology, risk assessment and management. Finnish Institute of Occupational Health, Helsinki, poster presentation at the 12th International Symposium on Dioxins and Related Compounds, Tampere, Finland

  • Clark G, Tritscher A, Bell D, Lucier G (1992b) Integrated approach for evaluating species and interindividual differences in responsiveness to dioxins and structural analogs. Environ Perspect 98: 125–132

    Article  CAS  Google Scholar 

  • Cook JC, Greenlee WF (1989) Characterization of a specific binding protein for 2,3,7,8-tetrachlorodibenzo-p-dioxin in human thymic epithelial cells. Mol Pharmacol 35: 713–719

    PubMed  CAS  Google Scholar 

  • Cook JC, Dold KM, Greenlee WF (1987) An in vitro model for studying the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin to human thymus. Toxicol Appl Pharmacol 89: 256–268

    Article  PubMed  CAS  Google Scholar 

  • Dean JH, Cornacoff JB, Luster MI (1990) Toxicity to the immune system: a review. In: Hadden JW, Szentivanyi A (eds) Immunopharmacology review, vol. 1. Plenum, New York, pp 377–408

    Google Scholar 

  • Dencker L, Hassouin E, D’Argy R, Alm G (1985) Fetal thymus organ culture as an in vitro model for the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin and its congeners. Mol Pharmacol 27: 133–140

    PubMed  CAS  Google Scholar 

  • Gaido KW, Leonard LS, Maness SC, Andersen ME (1991) TCDD-dependent regulation of gene expression in human keratinocytes. CIIT Activities 11: 1–10

    Google Scholar 

  • Hoffman RE, Stehr-Green PA, Webb KB, Evans RG, Knutsen AP, Schramm WF, Staake JF, Gibson BB, Steinberg KK (1986) Health effects of longterm exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. JAMA 255: 2031–2038

    Article  PubMed  CAS  Google Scholar 

  • Holsapple MP, Dooley RK, McNerney PJ, McCay JA (1986) Direct suppression of antibody responses by chlorinated dibenzodioxins in cultured spleen cells from (C57BL/6xC3H)F1 and DBA/2 mice. Immunopharmacology 12: 175–186

    Article  PubMed  CAS  Google Scholar 

  • Holsapple MP, Morris DL, Wood SC, Snyder NK (1991) 2,3,7,8-Tetrachlorodibenzo-p-dioxin-induced changes in immunocompetence: possible mechanisms. Annu Rev Pharmacol Toxicol 31: 73–100

    PubMed  CAS  Google Scholar 

  • Hong R, Taylor K, Abonour R (1989) Immune abnormalities with chronic TCDD exposure in Rhesus. Chem 18: 313–320

    Google Scholar 

  • Jennings AM, Wild G, Ward JD, Ward AM (1988) Immunological abnormalities 17 years after accidental exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. BMJ 45: 701–704

    CAS  Google Scholar 

  • Johnson ES (1992) Human exposure to 2,3,7,8-TCDD and risk of cancer. Crit Rev Toxicol 21: 451–463

    Article  CAS  Google Scholar 

  • Kawamoto T, Matsumura F (1989) Effects of TCDD on the EGF receptor of XB mouse keratinizing epithelial cells. J Biochem Toxicol 4: 173–183

    Article  PubMed  CAS  Google Scholar 

  • Kervliet NI, Brauner JA (1990) Flow cytometric analysis of lymphocyte subpopulations in the spleen and thymus of mice exposed to an acute immunosuppressive dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Environ Res 52: 146–154

    Article  Google Scholar 

  • Knutsen AP (1984) Immunologic effects of TCDD exposure in humans. Bull Environ Contam Toxicol 33: 673–681

    Article  PubMed  CAS  Google Scholar 

  • Kochman S, Cazabat JBA, Lavaud F, Lorton C, Rappe C (1986) Phenotypical dissection of immunoregulatory T cell subsets in human after furan exposure. Chem 15: 9–12

    Google Scholar 

  • Kouri RE, Ratrie III H, Atlas SA, Niwa A, Nebert DW (1974) Aryl hydrocarbon hydroxylase induction in human lymphocyte cultures bt 2,3,7,8-tetrachlordibenzo-p-dioxin. Life Sci 15: 1585–1595

    Article  PubMed  CAS  Google Scholar 

  • Krishan A (1975) Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol 66: 188–193

    Article  PubMed  CAS  Google Scholar 

  • Lorenzen A, Okey AB (1991) Detection and characterization of Ah receptor in tissues and cells from human tonsils. Toxicol Pharmacol 107: 203–214

    CAS  Google Scholar 

  • Lundberg K, Dencker L, Grönvik K-O (1990) Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treatment in vivo on thymocyte functions in mice after activation in vivo. Int J Immunopharmacol 12: 459–466

    Article  PubMed  CAS  Google Scholar 

  • Lundberg K, Dencker L, Grönvik K-O (1992) 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) inhibits the activation of antigen-specific T-cells in mice. Int J Immunopharmacol 14: 699–705

    Article  PubMed  CAS  Google Scholar 

  • Luster MI, Boorman GA, Dean JH, Harris MW, Luebke RW, Padarathsingh ML, Moore JA (1980) Examination of bone marrow, immunologic parameters and host susceptibility following prepostnatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Int J Immunopharmacol 2: 301–310

    Article  PubMed  CAS  Google Scholar 

  • Nebert DW (1989) The Ah locus: genetic differences in toxicity, cancer, mutation, and birth defects. CRC Crit Rev Toxicol 20: 153–174

    Article  CAS  Google Scholar 

  • Neubert R, Jacob-Müller U, Stahlmann R, Neubert D (1991) Polyhalogenated dibenzo-p-dioxins and dibenzofurans and the immune system. 1. Effects on peripheral lymphocyte subpopulations of a non-human primate (Callithrix jacchus) after treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Arch Toxicol 64: 345–359

    Article  Google Scholar 

  • Neubert R, Jacob-Müller U, Helge H, Stahlmann R, Neubert D (1991) Polyhalogenated dibenzo-p-dioxins and dibenzofurans and the immune system. 2. In vitro effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on lymphocytes of venous blood from man and a non-human primate (Callithrix jacchus). Arch Toxicol 65: 213–219

    Article  PubMed  CAS  Google Scholar 

  • Neubert R, Golor G, Stahlmann, R, Helge H, Neubert D (1992) Polyhalogenated dibenzo-p-dioxins and dibenzofurans and the immune system. 4. Effects of multiple-dose treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on peripheral lymphocyte subpopulations of a non-human primate (Callithrix jacchus). Arch Toxicol 66: 250–259

    Article  PubMed  CAS  Google Scholar 

  • Pohl RJ, Fouts JR (1980) A rapid method for assaying the metabolism of 7-ethoxyresorufin by microsomal subcellular fractions. Anal Biochem 107: 150–155

    Article  PubMed  CAS  Google Scholar 

  • Poiger H, Schlatter Ch (1983) Animal toxicology of chlorinated dibenzo-p-dioxins. Chem 12: 453–462

    Article  CAS  Google Scholar 

  • Puhvel SM, Sakamoto M (1988) Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on murine skin. J Invest Dermatol 30: 354–358

    Article  Google Scholar 

  • Silbergeld EK (1991) Dioxin: a case study in chloracne. In: Marzulli FN, Maibach HI (eds.) Dermatotoxicology. Hemisphere, New York, pp 667–686

    Google Scholar 

  • Silva JA, Kaufman CA, Simon DG et al. (1979) Lymphocyte function in humans exposed to polybrominated biphenyls. J Retic Soc 26: 341–347

    CAS  Google Scholar 

  • Taylor MJ, Lucier GW, Mahler JF, Thompson M, Lockhart AC, Clark GC (1992) Toxicol Appl Pharmacol 117: 126–132

    Article  PubMed  CAS  Google Scholar 

  • Tognoni G, Bonaccorsi A (1982) Epidemiological problems with TCDD (a critical review). Drug Metab Rev 13: 447–469

    Article  PubMed  CAS  Google Scholar 

  • Tomar RS, Kerkvliet NI (1991) Reduced T-helper cell function in mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Toxicol Lett 57: 55–64

    Article  PubMed  CAS  Google Scholar 

  • Tucker AN, Vore SJ, Luster MI (1986) Suppression of B cell differentiation by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Mol Pharmacol 29: 372–377

    PubMed  CAS  Google Scholar 

  • Whitlock Jr JP (1990) Genetic and molecular aspects of 2,3,7,8-tetrachlorodibenzo-p-dioxin action. Annu Rev Pharmacol Toxicol 30: 251–277

    PubMed  CAS  Google Scholar 

  • Wood SC, Karras JG, Holsapple MP (1992) Integration of the human lymphocyte into immunotoxicological investigations. Fundam Appl Toxicol 18: 450–459

    Article  PubMed  CAS  Google Scholar 

  • Wu Y-C, Hsieh R-P, Lü Y-C (1984) Altered distribution of lymphocyte subpopulations and augmentation of lymphocyte proliferation in chronic PCB poisoned patients. Chin J Microbiol Immunol 17: 177–187

    CAS  Google Scholar 

  • Zola H (1987) The surface antigens of human B lymphocytes. Immunol T Today 8: 308–315

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Center for Environmental Medicine and Lung Biology, University of North Carolina, Chapel Hill, NC, USA

    Dagmar S. Lang

  2. TRC-Environmental Corporation, Inc., Chapel Hill, NC, USA

    Susannne Becker

  3. Laboratory of Biochemical Risk Analysis, National Institute of Environmental Health Sciences, RTP, NC, USA

    George C. Clark

  4. Health Effects Research Laboratory, US Environmental Protection Agency, RTP, NC, USA

    Robert B. Devlin & Hillel S. Koren

Authors
  1. Dagmar S. Lang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susannne Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. George C. Clark
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Robert B. Devlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hillel S. Koren
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The research described in this article has been reviewed by the Health Effects Research Laboratory, US Environmental Protection Agency, and apporved for puplication. Approval, does not signify that the contents reflect the views and policies of the Agency, nor does mention of trade names or commercial products consitute endorsement of recommendation for use.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lang, D.S., Becker, S., Clark, G.C. et al. Lack of direct immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on human peripheral blood lymphocyte subsets in vitro. Arch Toxicol 68, 296–302 (1994). https://doi.org/10.1007/s002040050072

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s002040050072

Key words

Search

Navigation