ДИНАМИКА ЭКСПРЕССИИ ЦИТОКИНА IL-12 В МАКРОФАГАХ ЧЕЛОВЕКА ПОСЛЕ ИХ ОБРАБОТКИ ДИОКСИНОМ

Интерлейкин-12 (IL-12) является важнейшим провоспалительным цитокином, синтезируемым макрофагами, однако данные о влиянии диоксина на его экспрессию до сих пор фрагментарны. Наличие выявленных нами ранее потенциальных DRE (dioxin responsive elements) в регуляторных районах генов IL12A и IL12B, кодирующих субъединицы цитокина IL-12p35 и IL-12p40 соответственно, предполагает возможность прямой активации этих генов через связывание DRE с комплексом диоксин /AhR/ARNT.

В настоящей работе связывающая способность этих DRE доказана с помощью гель-шифт анализа. Исследование динамики экспрессии генов IL12A и IL12B на модели макрофагоподобных клеток человека линии U937 не выявило влияния диоксина на уровень экспрессии гена IL12A. В то же время выявлена кратковременная активация, а затем падение экспрессии гена IL12B. Наблюдаемая динамика может объясняться прямой активацией экспрессии диоксин-содержащим комплексом и последующим подавлением экспрессии в результате оксидативного стресса, вызываемого диоксином. Таким образом, известный факт влияния диоксина на иммунную систему может быть связан в том числе и с различным его влиянием на динамику экспрессии генов, кодирующих субъединицы IL-12.

1. Alsharif N.Z., Lawson T., Stohs S.J. Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin is mediated by the aryl hydrocarbon (Ah) receptor complex // Toxicology. 1994. V. 92. No. 1/3. P. 39–51.

2. Babik J.M., Adams E., Tone Y., Fairchild P.J., Tone M., Waldmann H. Expression of murine IL-12 is regulated by translational control of the p35 subunit // J. Immunol. 1999. V. 162. No. 7. P. 4069–4078.

3. Biswas S.K., Chittezhath M., Shalova I.N., Lim J.Y. Macrophage polarization and plasticity in health and disease // Immunol. Res. 2012. V. 53. No. 1/3. P. 11–24.

4. Boutros P.C., Moffat I.D., Franc M.A., Tuomisto J., Okey A.B. Dioxin-responsive AHRE-II gene battery: identification by phylogenetic footprinting // J. Bichem. Biophys. Res. Commun. 2004. V. 321. No. 3. P. 707–715.

5. Ciftci O., Tanyildizi S., Godekmerdan A. Protective effect of curcumin on immune system and body weight gain on rats intoxicated with 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) // Immunopharmacol. Immunotoxicol. 2010. V. 32. No. 1. P. 99–104.

6. Connor K.T., Aylward L.L. Human response to dioxin: aryl hydrocarbon receptor (AhR) molecular structure, function, and dose-response data for enzyme induction indicate an impaired human AhR // J. Toxicol. Environ. Health B Crit. Rev. 2006. V. 9. No. 2. P. 147–171.

7. Denison M.S., Nagy S.R. Activation of the aryl hydrocarbon receptor by structurally diverse exogenous and endogenous chemicals // Annu. Rev. Pharmacol. Toxicol. 2003. V. 43. P. 309–334.

8. Fujii-Kuriyama Y., Kawajiri K. Molecular mechanisms of the physiological functions of the aryl hydrocarbon (dioxin) receptor, a multifunctional regulator that senses and responds to environmental stimuli // Proc. Jpn. Acad. Ser. B Phys. Biol. Sci. 2010. V. 86. No. 1. P. 40–53.

9. Fujimaki H., Nohara K., Kobayashi T., Suzuki K., Eguchi-Kasai K., Tsukumo S., Kijima M., Tohyama C. Effect of a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin on immune function in male NC/Nga mice // Toxicol. Sci. 2002. V. 66. No. 1. P. 117–24.

10. Fujita H., Samejima H., Kitagawa N., Mitsuhashi T., Washio T., Yonemoto J., Tomita M., Takahashi T., Kosaki K. Genome-wide screening of dioxin-responsive genes in fetal brain: bioinformatic and experimental approaches // Cong. Anom. 2006. V. 46. P. 135–143.

11. Furman D.P., Oshchepkova E.A., Oshchepkov D.Yu., Shamanina M.Yu., Mordvinov V.A. Promoters of the genes encoding the transcription factors regulating the cytokine gene expression in macrophages contain putative binding sites for aryl hydrocarbon receptor // Comput. Biol. Chem. 2009. V. 33. No. 6. P. 465–468.

12. Gee K., Guzzo C., Che Mat N.F., Ma W., Kumar A. The IL-12 family of cytokines in infection, infl ammation and autoimmune disorders // Infl amm. Allergy Drug Targets. 2009. V. 8. No. 1. P. 40–52.

13. Gilchrist M., Thorsson V., Li B., Rust A.G., Korb M., Roach J.C., Kennedy K., Hai T., Bolouri H., Aderem A. Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4 // Nature. 2006. V. 441. No. 7090. P. 173–178.

14. Gordon S., Taylor P.R. Monocyte and macrophage heterogeneity // Nat. Rev. Immunol. 2005. V. 5. No. 12. P. 953–964.

15. Hamza T., Barnett J.B., Li B. Interleukin 12 a key immunoregulatory cytokine in infection applications // Int. J. Mol. Sci. 2010. V. 26. No. 11(3). P. 789–806.

16. Jalah R., Rosati M., Ganneru B., Pilkington G.R., Valentin A., Kulkarni V., Bergamaschi C., Chowdhury B., Zhang G.M., Beach R.K., Alicea C., Broderick K.E., Sardesai N.Y., Pavlakis G.N., Felber B.K. The p40 subunit of interleukin (IL)-12 promotes stabilization and export of the p35 subunit: implications for improved IL-12 cytokine production // J. Biol. Chem. 2013. V. 288. No. 9. P. 6763–6776.

17. Jin M.H., Hong C.H., Lee H.Y., Kang H.J., Han S.W. Enhanced TGF-beta1 is involved in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced oxidative stress in C57BL/6 mouse testis // Toxicol. Lett. 2008. V. 178. No. 3. P. 202–209.

18. Kerkvliet N.I. TCDD: an environmental immunotoxicant reveals a novel pathway of immunoregulation – a 30-year odyssey // Toxicol. Pathol. 2012. V. 40. No. 2. P. 138–142.

19. Knerr S., Schrenk D. Carcinogenicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in experimental models // Mol. Nutr. Food Res. 2006. V. 50. No. 10. P. 897–907.

20. Komura K., Hayashi S., Makino I., Poellinger L., Tanaka H. Aryl hydrocarbon receptor/dioxin receptor in human monocytes and macrophages // Mol. Cell Biochem. 2001. V. 226. No. 1/2. P. 107–118.

21. Mandal Prabir K. Dioxin: a review of its environmental effects and its aryl hydrocarbon receptor biology // J. Comp. Physiol. (B). 2005. V. 175. P. 221–230.

22. Marshall N.B., Kerkvliet N.I. Dioxin and immune regulation: emerging role of aryl hydrocarbon receptor in the generation of regulatory T cells // Ann. N.Y. Acad. Sci. 2010. V. 1183. P. 25–37.

23. Meyer L.R., Zweig A.S., Hinrichs A.S., Karolchik D., Kuhn R.M., Wong M., Sloan C.A., Rosenbloom K.R., Roe G., Rhead B., Raney B.J., Pohl A., Malladi V.S., Li C.H., Lee B.T., Learned K., Kirkup V., Hsu F., Heitner S., Harte R.A., Haeussler M., Guruvadoo L., Goldman M., Giardine B.M., Fujita P.A., Dreszer T.R., Diekhans M., Cline M.S., Clawson H., Barber G.P., Haussler D., Kent W.J. The UCSC Genome Browser database: extensions and updates 2013 // Nucl. Acids Res. 2013. V. 41. P. 64–69.

24. Neff-LaFord H., Teske S., Bushnell T.P., Lawrence B.P. Aryl hydrocarbon receptor activation during infl uenza virus infection unveils a novel pathway of IFN-gamma production by phagocytic cells // J. Immunol. 2007. V. 179. No. 1. P. 247–255.

25. Nguyen N.T., Hanieh H., Nakahama T., Kishimoto T. The roles of aryl hydrocarbon receptor in immune responses // Int. Immunol. 2013. V. 25. No. 6. P. 335–343.

26. Oshchepkov D.Y., Vityaev E.E., Grigorovich D.A., Ignatieva E.V., Khlebodarova T.M. SITECON: a tool for detecting conservative conformational and physicochemical properties in transcription factor binding site alignments and for site recognition // Nucl. Acids Res. 2004. V. 32. P. 208–212.

27. Raney B.J., Cline M.S., Rosenbloom K.R., Dreszer T.R., Learned K., Barber G.P., Meyer L.R., Sloan C.A., Malladi V. S., Roskin K.M., Suh B.B., Hinrichs A.S., Clawson H., Zweig A.S., Kirkup V., Fujita P.A., Rhead B., Smith K.E., Pohl A., Kuhn R.M., Karolchik D., Haussler D., Kent W.J. ENCODE whole-genome data in the UCSC genome browser (2011 update) // Nucl. Acids Res. 2011. V. 39. P. 871–875.

28. Schulz V.J., Smit J.J., Pieters R.H. The aryl hydrocarbon receptor and food allergy // Vet. Q. 2013. V. 33. No. 2. P. 94–107.

29. Sciullo E.M., Dong B., Vogel C.F., Matsumura F. Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early infl ammatory responses in U937 human macrophages // Chemosphere. 2009. V. 74. No. 11. P. 1531–1537.

30. Simones T., Shepherd D.M. Consequences of AhR activation in steady-state dendritic cells // Toxicol. Sci. 2011. V. 119. No. 2. P. 293–307.

31. Steenland K., Bertazzi P., Baccarelli A., Kogevinas M. Dioxin revisited: developments since the 1997 IARC classifi cation of dioxin as a human carcinogen // Environ. Health Perspect. 2004. V. 112. No. 13. P. 1265–1268.

32. Stohs S.J. Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) // Free Radic. Biol. Med. 1990. V. 9. No. 1. P. 79–90.

33. Sun Y.V., Boverhof D.R., Burgoon L.D., Fielden M.R., Zacharewski T.R. Comparative analysis of dioxin response elements in human, mouse and rat genomic sequences // Nucl. Acids Res. 2004. V. 32. P. 4512–4523.

34. Walker N.J., Portier C.J., Lax S.F., Crofts F.G., Li Y., Lucier G.W., Sutter T.R. Characterization of the dose-response of CYP1B1, CYP1A1, and CYP1A2 in the liver of female Sprague-Dawley rats following chronic exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin // Toxicol. Appl. Pharmacol. 1999. V. 154. P. 279–286.

35. Watford W.T., Moriguchi M., Morinobu A., O’Shea J.J. The biology of IL-12: coordinating innate and adaptive immune responses // Cytokine Growth Factor Rev. 2003. V. 14. No. 5. P. 361–368.

36. Zheng S., Abraham C. NF-κB1 inhibits NOD2-induced cytokine secretion through ATF3-dependent mechanisms // Mol. Cell. Biol. 2013. V. 33. No. 24. P. 4857–4871.

37. Zodrow J.M., Stegeman J.J., Tanguay R.L. Histological analysis of acute toxicity of 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD) in zebrafi sh // Aquat. Toxicol. 2004. V. 66. P. 25–38.