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. 2004 May;112(7):782–791. doi: 10.1289/ehp.112-1241993

Toxicogenomics of subchronic hexachlorobenzene exposure in Brown Norway rats.

Janine Ezendam 1, Frank Staedtler 1, Jeroen Pennings 1, Rob J Vandebriel 1, Raymond Pieters 1, Paolo Boffetta 1, Johannes H Harleman 1, Joseph G Vos 1
PMCID: PMC1241993  PMID: 15159207

Abstract

Hexachlorobenzene (HCB) is a persistent environmental pollutant with toxic effects in man and rat. Reported adverse effects are hepatic porphyria, neurotoxicity, and adverse effects on the reproductive and immune system. To obtain more insight into HCB-induced mechanisms of toxicity, we studied gene expression levels using DNA microarrays. For 4 weeks, Brown Norway rats were fed a diet supplemented with 0, 150, or 450 mg HCB/kg. Spleen, mesenteric lymph nodes (MLN), thymus, blood, liver, and kidney were collected and analyzed using the Affymetrix rat RGU-34A GeneChip microarray. Most significant (p < 0.001) changes, compared to the control group, occurred in spleen, followed by liver, kidney, blood, and MLN, but only a few genes were affected in thymus. This was to be expected, as the thymus is not a target organ of HCB. Transcriptome profiles confirmed known effects of HCB such as stimulatory effects on the immune system and induction of enzymes involved in drug metabolism, porphyria, and the reproductive system. In line with previous histopathological findings were increased transcript levels of markers for granulocytes and macrophages. New findings include the upregulation of genes encoding proinflammatory cytokines, antioxidants, acute phase proteins, mast cell markers, complements, chemokines, and cell adhesion molecules. Generally, gene expression data provide evidence that HCB induces a systemic inflammatory response, accompanied by oxidative stress and an acute phase response. In conclusion, this study confirms previously observed (immuno)toxicological effects of HCB but also reveals several new and mechanistically relevant gene products. Thus, transcriptome profiles can be used as markers for several of the processes that occur after HCB exposure.

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Selected References

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  1. Alvarez L., Randi A., Alvarez P., Piroli G., Chamson-Reig A., Lux-Lantos V., Kleiman de Pisarev D. Reproductive effects of hexachlorobenzene in female rats. J Appl Toxicol. 2000 Jan-Feb;20(1):81–87. doi: 10.1002/(sici)1099-1263(200001/02)20:1<81::aid-jat629>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  2. Bickers D. R. The dermatologic manifestations of human porphyria. Ann N Y Acad Sci. 1987;514:261–267. doi: 10.1111/j.1749-6632.1987.tb48781.x. [DOI] [PubMed] [Google Scholar]
  3. Bigby Timothy D. The yin and the yang of 5-lipoxygenase pathway activation. Mol Pharmacol. 2002 Aug;62(2):200–202. doi: 10.1124/mol.62.2.200. [DOI] [PubMed] [Google Scholar]
  4. Billi de Catabbi S., Sterin-Speziale N., Fernandez M. C., Minutolo C., Aldonatti C., San Martín de Viale L. Time course of hexachlorobenzene-induced alterations of lipid metabolism and their relation to porphyria. Int J Biochem Cell Biol. 1997 Feb;29(2):335–344. doi: 10.1016/s1357-2725(96)00096-9. [DOI] [PubMed] [Google Scholar]
  5. CAM C. [A new epidemic dermatosis of children]. Ann Dermatol Syphiligr (Paris) 1960 Jul-Aug;87:393–397. [PubMed] [Google Scholar]
  6. Chen Shangwu, Anderson Per O., Li Li, Sjögren Hans-Olov, Wang Ping, Li Su-Ling. Functional association of cytokine-induced SH2 protein and protein kinase C in activated T cells. Int Immunol. 2003 Mar;15(3):403–409. doi: 10.1093/intimm/dxg039. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Courtney K. D. Hexachlorobenzene (HCB): a review. Environ Res. 1979 Dec;20(2):225–266. doi: 10.1016/0013-9351(79)90001-x. [DOI] [PubMed] [Google Scholar]
  9. De Baets M., Stassen M. H. W. The role of antibodies in myasthenia gravis. J Neurol Sci. 2002 Oct 15;202(1-2):5–11. doi: 10.1016/s0022-510x(02)00200-9. [DOI] [PubMed] [Google Scholar]
  10. Dicou E., Perrot S., Menkes C. J., Masson C., Nerriere V. Nerve growth factor (NGF) autoantibodies and NGF in the synovial fluid: implications in spondylarthropathies. Autoimmunity. 1996;24(1):1–9. doi: 10.3109/08916939608995352. [DOI] [PubMed] [Google Scholar]
  11. Ezendam Janine, Hassing Ine, Bleumink Rob, Vos Joseph G., Pieters Raymond. Hexachlorobenzene-induced Immunopathology in Brown Norway rats is partly mediated by T cells. Toxicol Sci. 2003 Dec 2;78(1):88–95. doi: 10.1093/toxsci/kfh034. [DOI] [PubMed] [Google Scholar]
  12. Foster W. G., McMahon A., Younglai E. V., Jarrell J. F., Lecavalier P. Alterations in circulating ovarian steroids in hexachlorobenzene-exposed monkeys. Reprod Toxicol. 1995 Nov-Dec;9(6):541–548. doi: 10.1016/0890-6238(95)02004-7. [DOI] [PubMed] [Google Scholar]
  13. Franklin M. R., Phillips J. D., Kushner J. P. Cytochrome P450 induction, uroporphyrinogen decarboxylase depression, porphyrin accumulation and excretion, and gender influence in a 3-week rat model of porphyria cutanea tarda. Toxicol Appl Pharmacol. 1997 Dec;147(2):289–299. doi: 10.1006/taap.1997.8282. [DOI] [PubMed] [Google Scholar]
  14. Gocmen A., Peters H. A., Cripps D. J., Morris C. R., Dogramaci I. Porphyria turcica: hexachlorobenzene-induced porphyria. IARC Sci Publ. 1986;(77):567–573. [PubMed] [Google Scholar]
  15. Hahn M. E., Goldstein J. A., Linko P., Gasiewicz T. A. Interaction of hexachlorobenzene with the receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin in vitro and in vivo. Evidence that hexachlorobenzene is a weak Ah receptor agonist. Arch Biochem Biophys. 1989 Apr;270(1):344–355. doi: 10.1016/0003-9861(89)90037-4. [DOI] [PubMed] [Google Scholar]
  16. Hamadeh Hisham K., Bushel Pierre R., Jayadev Supriya, DiSorbo Olimpia, Bennett Lee, Li Leping, Tennant Raymond, Stoll Raymond, Barrett J. Carl, Paules Richard S. Prediction of compound signature using high density gene expression profiling. Toxicol Sci. 2002 Jun;67(2):232–240. doi: 10.1093/toxsci/67.2.232. [DOI] [PubMed] [Google Scholar]
  17. Hamadeh Hisham K., Bushel Pierre R., Jayadev Supriya, Martin Karla, DiSorbo Olimpia, Sieber Stella, Bennett Lee, Tennant Raymond, Stoll Raymond, Barrett J. Carl. Gene expression analysis reveals chemical-specific profiles. Toxicol Sci. 2002 Jun;67(2):219–231. doi: 10.1093/toxsci/67.2.219. [DOI] [PubMed] [Google Scholar]
  18. Hühn P., Pruijn G. J., van Venrooij W. J., Bachmann M. Characterization of the autoantigen La (SS-B) as a dsRNA unwinding enzyme. Nucleic Acids Res. 1997 Jan 15;25(2):410–416. doi: 10.1093/nar/25.2.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jarrell J. F., Gocmen A., Akyol D., Brant R. Hexachlorobenzene exposure and the proportion of male births in Turkey 1935-1990. Reprod Toxicol. 2002 Jan-Feb;16(1):65–70. doi: 10.1016/s0890-6238(01)00196-4. [DOI] [PubMed] [Google Scholar]
  20. Jarrell J., Gocmen A., Foster W., Brant R., Chan S., Sevcik M. Evaluation of reproductive outcomes in women inadvertently exposed to hexachlorobenzene in southeastern Turkey in the 1950s. Reprod Toxicol. 1998 Jul-Aug;12(4):469–476. doi: 10.1016/s0890-6238(98)00019-7. [DOI] [PubMed] [Google Scholar]
  21. Kennedy S. W., Wigfield D. C. Dose-response relationships in hexachlorobenzene-induced porphyria. Biochem Pharmacol. 1990 Sep 15;40(6):1381–1388. doi: 10.1016/0006-2952(90)90407-c. [DOI] [PubMed] [Google Scholar]
  22. Kleiman de Pisarev D. L., Rios de Molina M. C., San Martin de Viale L. C. Thyroid function and thyroxine metabolism in hexachlorobenzene-induced porphyria. Biochem Pharmacol. 1990 Mar 1;39(5):817–825. doi: 10.1016/0006-2952(90)90195-q. [DOI] [PubMed] [Google Scholar]
  23. Liu Guoying, Loraine Ann E., Shigeta Ron, Cline Melissa, Cheng Jill, Valmeekam Venu, Sun Shaw, Kulp David, Siani-Rose Michael A. NetAffx: Affymetrix probesets and annotations. Nucleic Acids Res. 2003 Jan 1;31(1):82–86. doi: 10.1093/nar/gkg121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lockhart D. J., Dong H., Byrne M. C., Follettie M. T., Gallo M. V., Chee M. S., Mittmann M., Wang C., Kobayashi M., Horton H. Expression monitoring by hybridization to high-density oligonucleotide arrays. Nat Biotechnol. 1996 Dec;14(13):1675–1680. doi: 10.1038/nbt1296-1675. [DOI] [PubMed] [Google Scholar]
  25. Machala M., Mátlová L., Svoboda I., Nezveda K. Induction effects of polychlorinated biphenyls, polycyclic aromatic hydrocarbons and other widespread aromatic environmental pollutants on microsomal monooxygenase activities in chick embryo liver. Arch Toxicol. 1996;70(6):362–367. doi: 10.1007/s002040050286. [DOI] [PubMed] [Google Scholar]
  26. Mantovani A., Allavena P., Vecchi A., Sozzani S. Chemokines and chemokine receptors during activation and deactivation of monocytes and dendritic cells and in amplification of Th1 versus Th2 responses. Int J Clin Lab Res. 1998;28(2):77–82. doi: 10.1007/s005990050023. [DOI] [PubMed] [Google Scholar]
  27. Michielsen C. C., van Loveren H., Vos J. G. The role of the immune system in hexachlorobenzene-induced toxicity. Environ Health Perspect. 1999 Oct;107 (Suppl 5):783–792. doi: 10.1289/ehp.99107s5783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Michielsen C. P., Bloksma N., Ultee A., van Mil F., Vos J. G. Hexachlorobenzene-induced immunomodulation and skin and lung lesions: a comparison between brown Norway, Lewis, and Wistar rats. Toxicol Appl Pharmacol. 1997 May;144(1):12–26. doi: 10.1006/taap.1997.8104. [DOI] [PubMed] [Google Scholar]
  29. Michielsen C., Zeamari S., Leusink-Muis A., Vos J., Bloksma N. The environmental pollutant hexachlorobenzene causes eosinophilic and granulomatous inflammation and in vitro airways hyperreactivity in the Brown Norway rat. Arch Toxicol. 2002 Mar 13;76(4):236–247. doi: 10.1007/s00204-002-0326-x. [DOI] [PubMed] [Google Scholar]
  30. Naka Tetsuji, Nishimoto Norihiro, Kishimoto Tadamitsu. The paradigm of IL-6: from basic science to medicine. Arthritis Res. 2002 May 9;4 (Suppl 3):S233–S242. doi: 10.1186/ar565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perretti Mauro, Gavins Felicity N. E. Annexin 1: an endogenous anti-inflammatory protein. News Physiol Sci. 2003 Apr;18:60–64. doi: 10.1152/nips.01424.2002. [DOI] [PubMed] [Google Scholar]
  32. Queiroz M. L., Bincoletto C., Perlingeiro R. C., Quadros M. R., Souza C. A. Immunoglobulin levels in workers exposed to hexachlorobenzene. Hum Exp Toxicol. 1998 Mar;17(3):172–175. doi: 10.1177/096032719801700308. [DOI] [PubMed] [Google Scholar]
  33. Queiroz M. L., Quadros M. R., Valadares M. C., Silveira J. P. Polymorphonuclear phagocytosis and killing in workers occupationally exposed to hexachlorobenzene. Immunopharmacol Immunotoxicol. 1998 Aug;20(3):447–454. doi: 10.3109/08923979809034826. [DOI] [PubMed] [Google Scholar]
  34. Renner G. Biotransformation of the fungicides hexachlorobenzene and pentachloronitrobenzene. Xenobiotica. 1981 Jul;11(7):435–446. doi: 10.3109/00498258109045854. [DOI] [PubMed] [Google Scholar]
  35. Roth Johannes, Vogl Thomas, Sorg Clemens, Sunderkötter Cord. Phagocyte-specific S100 proteins: a novel group of proinflammatory molecules. Trends Immunol. 2003 Apr;24(4):155–158. doi: 10.1016/s1471-4906(03)00062-0. [DOI] [PubMed] [Google Scholar]
  36. Schielen P., Schoo W., Tekstra J., Oostermeijer H. H., Seinen W., Bloksma N. Autoimmune effects of hexachlorobenzene in the rat. Toxicol Appl Pharmacol. 1993 Oct;122(2):233–243. doi: 10.1006/taap.1993.1192. [DOI] [PubMed] [Google Scholar]
  37. Schreck R., Albermann K., Baeuerle P. A. Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryotic cells (a review). Free Radic Res Commun. 1992;17(4):221–237. doi: 10.3109/10715769209079515. [DOI] [PubMed] [Google Scholar]
  38. Schulte Agnes, Althoff Jürgen, Ewe Sigrid, Richter-Reichhelm Hans-Bernhard, BGVV Group Investigators Two immunotoxicity ring studies according to OECD TG 407-comparison of data on cyclosporin A and hexachlorobenzene. Regul Toxicol Pharmacol. 2002 Aug;36(1):12–21. doi: 10.1006/rtph.2002.1568. [DOI] [PubMed] [Google Scholar]
  39. Stonard M. D., Poli G., De Matteis F. Stimulation of liver heme oxygenase in hexachlorobenzene-induced hepatic porphyria. Arch Toxicol. 1998 May;72(6):355–361. doi: 10.1007/s002040050514. [DOI] [PubMed] [Google Scholar]
  40. Thomas Russell S., Rank David R., Penn Sharron G., Zastrow Gina M., Hayes Kevin R., Hu Tianhua, Pande Kalyan, Lewis Mark, Jovanovich Stevan B., Bradfield Christopher A. Application of genomics to toxicology research. Environ Health Perspect. 2002 Dec;110 (Suppl 6):919–923. doi: 10.1289/ehp.02110s6919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Van Birgelen A. P., Van der Kolk J., Fase K. M., Bol I., Poiger H., Brouwer A., Van den Berg M. Subchronic dose-response study of 2,3,7,8-tetrachlorodibenzo-p-dioxin in female Sprague-Dawley rats. Toxicol Appl Pharmacol. 1995 May;132(1):1–13. doi: 10.1006/taap.1995.1080. [DOI] [PubMed] [Google Scholar]
  42. Vos J. G. Health effects of hexachlorobenzene and the TEF approach. Environ Health Perspect. 2000 Feb;108(2):A58–A58. doi: 10.1289/ehp.108-a58a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vos J. G. Immunotoxicity of hexachlorobenzene. IARC Sci Publ. 1986;(77):347–356. [PubMed] [Google Scholar]
  44. Vos J. G., van Logten M. J., Kreeftenberg J. G., Kruizinga W. Hexachlorobenzene-induced stimulation of the humoral immune response in rats. Ann N Y Acad Sci. 1979 May 31;320:535–550. [PubMed] [Google Scholar]
  45. van Birgelen A. P. Hexachlorobenzene as a possible major contributor to the dioxin activity of human milk. Environ Health Perspect. 1998 Nov;106(11):683–688. doi: 10.1289/ehp.106-1533492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. van Ommen B., van Bladeren P. J. Possible reactive intermediates in the oxidative biotransformation of hexachlorobenzene. Drug Metabol Drug Interact. 1989;7(2-3):213–243. doi: 10.1515/dmdi.1989.7.2-3.213. [DOI] [PubMed] [Google Scholar]

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