[go: up one dir, main page]
More Web Proxy on the site http://driver.im/
Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Obesity in C57BL/6J mice is characterized by adipose tissue hypoxia and cytotoxic T-cell infiltration

International Journal of Obesity volume 32, pages 451–463 (2008)Cite this article

Abstract

Background:

Obesity is currently viewed as a state of chronic low-grade inflammation in which there is a pro-inflammatory alteration in the serum adipocytokine profile as well as an infiltration of white adipose tissue by activated macrophages. The etiology of this inflammation, however, is poorly understood.

Methods:

Hypothesizing that local hypoxia within expanding white adipose tissue depots may contribute to obesity-related inflammation, we compared body composition, serum inflammatory marker concentrations and the expression of several hypoxia-regulated genes in white adipose tissue derived from lean, dietary-induced obese (DIO) and ob/ob male C57BL/6J mice. We also examined white adipose tissue for the presence of hypoxia using both a pimonidazole-based antibody system and a fiberoptic sensor for real-time pO2 quantification in vivo. Finally, using cell-specific leukocyte antibodies, we performed immunohistochemistry and flow cytometric analyses to further characterize the cellular nature of adipose inflammation.

Results:

We determined that obesity in male C57BL/6J mice is associated with increased expression of HIF (hypoxia-inducible factor) isoforms and GLUT-1, and that white adipose tissue hypoxia was present in the obese mice. Immunohistochemistry revealed hypoxic areas to colocalize predominantly with F4/80+ macrophages. Interestingly, CD3+ T cells were present in large numbers within the adipose of both DIO and ob/ob obese mice, and flow cytometry revealed their adipose to possess significantly more CD8+ T cells than their lean cohort.

Conclusions:

White adipose hypoxia and cytotoxic T-cell invasion are features of obesity in C57BL/6J mice and are potential contributors to their local and generalized inflammatory state.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10

Similar content being viewed by others

References

  1. Mendez MA, Monteiro CA, Popkin BM . Overweight exceeds underweight among women in most developing countries. Am J Clin Nutr 2005; 81: 714–721.

    Article  CAS  PubMed  Google Scholar 

  2. Rigby NJ, Kumanyika S, James WP . Confronting the epidemic: the need for global solutions. J Public Health Policy 2004; 25: 418–434.

    Article  PubMed  Google Scholar 

  3. Pi-Sunyer FX . The obesity epidemic: pathophysiology and consequences of obesity. Obes Res 2002; 10 (Suppl 2): 97S–104S.

    Article  PubMed  Google Scholar 

  4. Mokdad AH, Bowman BA, Ford ES, Vinicor F, Marks JS, Koplan JP . The continuing epidemics of obesity and diabetes in the United States. JAMA 2001; 286: 1195–1200.

    Article  CAS  PubMed  Google Scholar 

  5. Mokdad AH, Ford ES, Bowman BA, Dietz WH, Vinicor F, Bales VS et al. Prevalence of obesity, diabetes, and obesity-related health risk factors, 2001. JAMA 2003; 289: 76–79.

    Article  PubMed  Google Scholar 

  6. Speiser PW, Rudolf MC, Anhalt H, Camacho-Hubner C, Chiarelli F, Eliakim A et al. Childhood obesity. J Clin Endocrinol Metab 2005; 90: 1871–1887.

    Article  CAS  PubMed  Google Scholar 

  7. Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante Jr AW . Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest 2003; 112: 1796–1808.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Berg AH, Scherer PE . Adipose tissue, inflammation, and cardiovascular disease. Circ Res 2005; 96: 939–949.

    Article  CAS  PubMed  Google Scholar 

  9. Browning JD, Horton JD . Molecular mediators of hepatic steatosis and liver injury. J Clin Invest 2004; 114: 147–152.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Wellen KE, Hotamisligil GS . Inflammation, stress, and diabetes. J Clin Invest 2005; 115: 1111–1119.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Pickup JC . Inflammation and activated innate immunity in the pathogenesis of type 2 diabetes. Diabetes Care 2004; 27: 813–823.

    Article  PubMed  Google Scholar 

  12. Hotamisligil GS, Shargill NS, Spiegelman BM . Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science 1993; 259: 87–91.

    Article  CAS  PubMed  Google Scholar 

  13. Bastard JP, Jardel C, Delattre J, Hainque B, Bruckert E, Oberlin F . Evidence for a link between adipose tissue interleukin-6 content and serum C-reactive protein concentrations in obese subjects. Circulation 1999; 99: 2221–2222.

    Article  CAS  PubMed  Google Scholar 

  14. Fried SK, Bunkin DA, Greenberg AS . Omental and subcutaneous adipose tissues of obese subjects release interleukin-6: depot difference and regulation by glucocorticoid. J Clin Endocrinol Metab 1998; 83: 847–850.

    CAS  PubMed  Google Scholar 

  15. Lin HZ, Yang SQ, Chuckaree C, Kuhajda F, Ronnet G, Diehl AM . Metformin reverses fatty liver disease in obese, leptin-deficient mice. Nat Med 2000; 6: 998–1003.

    Article  CAS  PubMed  Google Scholar 

  16. Cai D, Yuan M, Frantz DF, Melendez PA, Hansen L, Lee J et al. Local and systemic insulin resistance resulting from hepatic activation of IKK-beta and NF-kappaB. Nat Med 2005; 11: 183–190.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Brodmerkel CM, Huber R, Covington M, Diamond S, Hall L, Collins R et al. Discovery and pharmacological characterization of a novel rodent-active CCR2 antagonist, INCB3344. J Immunol 2005; 175: 5370–5378.

    Article  CAS  PubMed  Google Scholar 

  18. Skurk T, Hauner H . Obesity and impaired fibrinolysis: role of adipose production of plasminogen activator inhibitor-1. Int J Obes Relat Metab Disord 2004; 28: 1357–1364.

    Article  CAS  PubMed  Google Scholar 

  19. Arkan MC, Hevener AL, Greten FR, Maeda S, Li ZW, Long JM et al. IKK-beta links inflammation to obesity-induced insulin resistance. Nat Med 2005; 11: 191–198.

    Article  CAS  PubMed  Google Scholar 

  20. Hirosumi J, Tuncman G, Chang L, Gorgun CZ, Uysal KT, Maeda K et al. A central role for JNK in obesity and insulin resistance. Nature 2002; 420: 333–336.

    Article  CAS  PubMed  Google Scholar 

  21. Yuan M, Konstantopoulos N, Lee J, Hansen L, Li ZW, Karin M et al. Reversal of obesity- and diet-induced insulin resistance with salicylates or targeted disruption of Ikkbeta. Science 2001; 293: 1673–1677.

    Article  CAS  PubMed  Google Scholar 

  22. Curat CA, Miranville A, Sengenes C, Diehl M, Tonus C, Busse R et al. From blood monocytes to adipose tissue-resident macrophages: induction of diapedesis by human mature adipocytes. Diabetes 2004; 53: 1285–1292.

    Article  CAS  PubMed  Google Scholar 

  23. Xu H, Barnes GT, Yang Q, Tan G, Yang D, Chou CJ et al. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J Clin Invest 2003; 112: 1821–1830.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Semenza GL . HIF-1: mediator of physiological and pathophysiological responses to hypoxia. J Appl Physiol 2000; 88: 1474–1480.

    Article  CAS  PubMed  Google Scholar 

  25. Griffiths JR, Robinson SP . The OxyLite: a fibre-optic oxygen sensor. Br J Radiol 1999; 72: 627–630.

    Article  CAS  PubMed  Google Scholar 

  26. Alessi MC, Juhan-Vague I . Contribution of PAI-1 in cardiovascular pathology. Arch Mal Coeur Vaiss 2004; 97: 673–678.

    CAS  PubMed  Google Scholar 

  27. Hoogeveen RC, Morrison A, Boerwinkle E, Miles JS, Rhodes CE, Sharrett AR et al. Plasma MCP-1 level and risk for peripheral arterial disease and incident coronary heart disease: Atherosclerosis Risk in Communities study. Atherosclerosis 2005; 183: 301–307.

    Article  CAS  PubMed  Google Scholar 

  28. Silha JV, Krsek M, Skrha JV, Sucharda P, Nyomba BL, Murphy LJ . Plasma resistin, adiponectin and leptin levels in lean and obese subjects: correlations with insulin resistance. Eur J Endocrinol 2003; 149: 331–335.

    Article  CAS  PubMed  Google Scholar 

  29. Garg A . Adipose tissue dysfunction in obesity and lipodystrophy. Clin Cornerstone 2006; 8 (Suppl 4): S7–S13.

    Article  PubMed  Google Scholar 

  30. Gleadle JM, Ratcliffe PJ . Induction of hypoxia-inducible factor-1, erythropoietin, vascular endothelial growth factor, and glucose transporter-1 by hypoxia: evidence against a regulatory role for Src kinase. Blood 1997; 89: 503–509.

    CAS  PubMed  Google Scholar 

  31. Zhou JT, Cai ZM, Li NC, Na YQ . Expression of hypoxia inducible factor-1alpha and glucose transporter protein 1 in renal and bladder cancers and the clinical significance thereof. Zhonghua Yi Xue Za Zhi 2006; 86: 1970–1974.

    CAS  PubMed  Google Scholar 

  32. Yaromina A, Zips D, Thames HD, Eicheler W, Krause M, Rosner A et al. Pimonidazole labelling and response to fractionated irradiation of five human squamous cell carcinoma (hSCC) lines in nude mice: The need for a multivariate approach in biomarker studies. Radiother Oncol 2006; 81: 122–129.

    Article  CAS  PubMed  Google Scholar 

  33. Tanaka T, Kato H, Kojima I, Ohse T, Son D, Tawakami T et al. Hypoxia and expression of hypoxia-inducible factor in the aging kidney. J Gerontol A Biol Sci Med Sci 2006; 61: 795–805.

    Article  PubMed  Google Scholar 

  34. Troost EG, Laverman P, Kaanders JH, Philippens M, Lok J, Oyen WJ et al. Imaging hypoxia after oxygenation-modification: comparing [18F]FMISO autoradiography with pimonidazole immunohistochemistry in human xenograft tumors. Radiother Oncol 2006; 80: 157–164.

    Article  CAS  PubMed  Google Scholar 

  35. Nordsmark M, Loncaster J, Aquino-Parsons C, Chou SC, Gebski V, West C et al. The prognostic value of pimonidazole and tumour pO2 in human cervix carcinomas after radiation therapy: a prospective international multi-center study. Radiother Oncol 2006; 80: 123–131.

    Article  CAS  PubMed  Google Scholar 

  36. Jankovic B, Aquino-Parsons C, Raleigh JA, Stanbridge EJ, Durand RE, Banath JP et al. Comparison between pimonidazole binding, oxygen electrode measurements, and expression of endogenous hypoxia markers in cancer of the uterine cervix. Cytometry B Clin Cytom 2006; 70: 45–55.

    Article  CAS  PubMed  Google Scholar 

  37. Urano M, Chen Y, Humm J, Koutcher JA, Zanzonico P, Ling C . Measurements of tumor tissue oxygen tension using a time-resolved luminescence-based optical oxylite probe: comparison with a paired survival assay. Radiat Res 2002; 158: 167–173.

    Article  CAS  PubMed  Google Scholar 

  38. Braun RD, Lanzen JL, Snyder SA, Dewhirst MW . Comparison of tumor and normal tissue oxygen tension measurements using OxyLite or microelectrodes in rodents. Am J Physiol Heart Circ Physiol 2001; 280: H2533–H2544.

    Article  CAS  PubMed  Google Scholar 

  39. Wen B, Urano M, O'Donoghue JA, Ling CC . Measurements of partial oxygen pressure pO2 using the OxyLite system in R3327-AT tumors under isoflurane anesthesia. Radiat Res 2006; 166: 512–518.

    Article  CAS  PubMed  Google Scholar 

  40. Tilg H, Moschen AR . Adipocytokines: mediators linking adipose tissue, inflammation and immunity. Nat Rev Immunol 2006; 6: 772–783.

    Article  CAS  PubMed  Google Scholar 

  41. Loffreda S, Yang SQ, Lin HZ, Karp CL, Brengman ML, Wang DJ et al. Leptin regulates proinflammatory immune responses. FASEB J 1998; 12: 57–65.

    Article  CAS  PubMed  Google Scholar 

  42. Grosfeld A, Andre J, Hauguel-De Mouzon S, Berra E, Pouyssegur J, Guerre-Millo M . Hypoxia-inducible factor 1 transactivates the human leptin gene promoter. J Biol Chem 2002; 277: 42953–42957.

    Article  CAS  PubMed  Google Scholar 

  43. Tsatsanis C, Zacharioudaki V, Androulidaki A, Dermitzaki E, Charalampopoulos I, Minas V et al. Adiponectin induces TNF-alpha and IL-6 in macrophages and promotes tolerance to itself and other pro-inflammatory stimuli. Biochem Biophys Res Commun 2005; 335: 1254–1263.

    Article  CAS  PubMed  Google Scholar 

  44. Mertens I, Verrijken A, Michiels JJ, Van der Planken M, Ruige JB, Van Gaal LF . Among inflammation and coagulation markers, PAI-1 is a true component of the metabolic syndrome. Int J Obes (London) 2006; 30: 1308–1314.

    Article  CAS  Google Scholar 

  45. Tortoriello DV, McMinn JE, Chua SC . Increased expression of hypothalamic leptin receptor and adiponectin accompany resistance to dietary-induced obesity and infertility in female C57BL/6J mice. Int J Obes (London) 2006; 31: 395–402.

    Article  Google Scholar 

  46. Weisberg SP, Hunter D, Huber R, Lemieux J, Slaymaker S, Vaddi K et al. CCR2 modulates inflammatory and metabolic effects of high-fat feeding. J Clin Invest 2005; 116: 115–124.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Cancello R, Henegar C, Viguerie N, Taleb S, Poitou C, Rouault C et al. Reduction of macrophage infiltration and chemoattractant gene expression changes in white adipose tissue of morbidly obese subjects after surgery-induced weight loss. Diabetes 2005; 54: 2277–2286.

    Article  CAS  PubMed  Google Scholar 

  48. Kim YM, An JJ, Jin YJ, Rhee Y, Cha BS, Lee HC et al. Assessment of the anti-obesity effects of the TNP-470 analog, CKD-732. J Mol Endocrinol 2007; 38: 455–465.

    Article  CAS  PubMed  Google Scholar 

  49. Cinti S, Mitchell G, Barbatelli G, Murano I, Ceresi E, Faloia E et al. Adipocyte death defines macrophage localization and function in adipose tissue of obese mice and humans. J Lipid Res 2005; 46: 2347–2355.

    Article  CAS  PubMed  Google Scholar 

  50. Helmlinger G, Yuan F, Dellian M, Jain RK . Interstitial pH and pO2 gradients in solid tumors in vivo: high-resolution measurements reveal a lack of correlation. Nat Med 1997; 3: 177–182.

    Article  CAS  PubMed  Google Scholar 

  51. Brook CG, Lloyd JK, Wolf OH . Relation between age of onset of obesity and size and number of adipose cells. BMJ 1972; 2: 25–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Bianciardi P, Fantacci M, Caretti A, Ronchi R, Milano G, Morel S et al. Chronic in vivo hypoxia in various organs: hypoxia-inducible factor-1alpha and apoptosis. Biochem Biophys Res Commun 2006; 342: 875–880.

    Article  CAS  PubMed  Google Scholar 

  53. Haase VH . Hypoxia-inducible factors in the kidney. Am J Physiol Renal Physiol 2006; 291: F271–F281.

    Article  CAS  PubMed  Google Scholar 

  54. Papandreou I, Cairns RA, Fontana L, Lim AL, Denko NC . HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. Cell Metab 2006; 3: 187–197.

    Article  CAS  PubMed  Google Scholar 

  55. Burke B, Giannoudis A, Corke KP, Gill D, Wells M, Ziegler-Heitbrock L et al. Hypoxia-induced gene expression in human macrophages: implications for ischemic tissues and hypoxia-regulated gene therapy. Am J Pathol 2003; 163: 1233–1243.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Cramer T, Yamanishi Y, Clausen BE, Forster I, Pawlinski R, Mackman N et al. HIF-1alpha is essential for myeloid cell-mediated inflammation. Cell 2003; 112: 645–657.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Lewis JS, Landers RJ, Underwood JC, Harris AL, Lewis CE . Expression of vascular endothelial growth factor by macrophages is up-regulated in poorly vascularized areas of breast carcinomas. J Pathol 2000; 192: 150–158.

    Article  CAS  PubMed  Google Scholar 

  58. Leek RD, Landers RJ, Harris AL, Lewis CE . Necrosis correlates with high vascular density and focal macrophage infiltration in invasive carcinoma of the breast. Br J Cancer 1999; 79: 991–995.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Jarpe AJ, Hickman MR, Anderson JT, Winter WE, Peck AB . Flow cytometric enumeration of mononuclear cell populations infiltrating the islets of Langerhans in prediabetic NOD mice: development of a model of autoimmune insulitis for type I diabetes. Reg Immunol 1990; 3: 305–317.

    PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by a Women's Reproductive Health Research Scholarship, NIH 5 K12 HD001275-04 (DVT).

Author information

Authors and Affiliations

  1. Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Columbia University, New York, NY, USA

    M E Rausch, P Vardhana & D V Tortoriello

  2. Division of Molecular Genetics, Department of Pediatrics, Columbia University, New York, NY, USA

    S Weisberg & D V Tortoriello

Authors
  1. M E Rausch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Weisberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P Vardhana
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D V Tortoriello
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D V Tortoriello.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rausch, M., Weisberg, S., Vardhana, P. et al. Obesity in C57BL/6J mice is characterized by adipose tissue hypoxia and cytotoxic T-cell infiltration. Int J Obes 32, 451–463 (2008). https://doi.org/10.1038/sj.ijo.0803744

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.ijo.0803744

Keywords

Search

Advanced search

Quick links