Background/Aims Regulatory T cells (Tregs) and natural killer T (NKT) cells are two distinct lymphocyte subsets that independently regulate hepatic adaptive and innate immunity, respectively. In the current study, we examine the interaction between Tregs and NKT cells to understand the mechanisms of cross immune regulation by these cells. Methods The frequency and function of Tregs were evaluated in wild type and NKT cell deficient (CD1dko) mice. In vitro lymphocyte proliferation and apoptosis assays were performed with NKT cells co-cultured with Tregs. The ability of Tregs to inhibit NKT cells in vivo was examined by adoptive transfer of Tregs in a model of NKT cell mediated hepatitis. Results CD1dko mice have a significant reduction in hepatic Tregs. Although, the Tregs from CD1dko mice remain functional and can suppress conventional T cells, their ability to suppress activation induced NKT cell proliferation and to promote NKT cell apoptosis is greatly diminished. These effects are CD1d dependent and require cell to cell contact. Adoptive transfer of Tregs inhibits NKT cell-mediated liver injury. Conclusions NKT cells promote Tregs, and Tregs inhibit NKT cells in a CD1d dependent manner requiring cell to cell contact. These cross-talk immune regulations provide a linkage between innate and adaptive immunity.
References
[1]
Qian S, Demetris AJ, Murase N, Rao AS, Fung JJ, et al. (1994) Murine liver allograft transplantation: tolerance and donor cell chimerism. Hepatology 19: 916–924.
[2]
Sakaguchi S, Powrie F (2007) Emerging Challenges in Regulatory T Cell Function and Biology. Science 317: 627–629.
[3]
Taniguchi M, Harada M, Kojo S, Nakayama T, Wakao H (2003) The regulatory role of Valpha14 NKT cells in innate and acquired immune response. Annu Rev Immunol 21: 483–513.
[4]
Jiang S, Lechler RI (2003) Regulatory T cells in the control of transplantation tolerance and autoimmunity. Am J Transplant 3: 516–524.
[5]
Cabrera R, Tu Z, Xu Y, Firpi RJ, Rosen HR, et al. (2004) An immunomodulatory role for CD4(+)CD25(+) regulatory T lymphocytes in hepatitis C virus infection. Hepatology 40: 1062–1071.
[6]
Longhi MS, Ma Y, Bogdanos DP, Cheeseman P, Mieli-Vergani G, et al. (2004) Impairment of CD4(+)CD25(+) regulatory T-cells in autoimmune liver disease. J Hepatol 41: 31–37.
[7]
Unitt E, Rushbrook SM, Marshall A, Davies S, Gibbs P, et al. (2005) Compromised lymphocytes infiltrate hepatocellular carcinoma: the role of T-regulatory cells. Hepatology 41: 722–730.
[8]
Hori S, Nomura T, Sakaguchi S (2003) Control of Regulatory T Cell Development by the Transcription Factor Foxp3. Science 299: 1057–1061.
[9]
Stoop JN, van der Molen RG, Baan CC, van der Laan LJ, Kuipers EJ, et al. (2005) Regulatory T cells contribute to the impaired immune response in patients with chronic hepatitis B virus infection. Hepatology 41: 771–778.
[10]
Lan RY, Cheng C, Lian ZX, Tsuneyama K, Yang GX, et al. (2006) Liver-targeted and peripheral blood alterations of regulatory T cells in primary biliary cirrhosis. Hepatology 43: 729–737.
[11]
Demirkiran A, Kok A, Kwekkeboom J, Kusters JG, Metselaar HJ, et al. (2006) Low circulating regulatory T-cell levels after acute rejection in liver transplantation. Liver Transpl 12: 277–284.
[12]
Kawano T, Cui J, Koezuka Y, Toura I, Kaneko Y, et al. (1997) CD1d-restricted and TCR-mediated activation of valpha14 NKT cells by glycosylceramides. Science 278: 1626–1629.
[13]
Godfrey DI, Stankovic S, Baxter AG (2010) Raising the NKT cell family. Nat Immunol 11: 197–206.
[14]
Godfrey DI, MacDonald HR, Kronenberg M, Smyth MJ, Van Kaer L (2004) NKT cells: what's in a name? Nat Rev Immunol 4: 231–237.
[15]
Klugewitz K, Adams DH, Emoto M, Eulenburg K, Hamann A (2004) The composition of intrahepatic lymphocytes: shaped by selective recruitment? Trends Immunol 25: 590–594.
[16]
Kronenberg M (2005) Toward an understanding of NKT cell biology: progress and paradoxes. Annu Rev Immunol 23: 877–900.
[17]
Ma X, Hua J, Li Z (2008) Probiotics improve high fat diet-induced hepatic steatosis and insulin resistance by increasing hepatic NKT cells. J Hepatol 49: 821–830.
[18]
Li Z, Soloski MJ, Diehl AM (2005) Dietary factors alter hepatic innate immune system in mice with nonalcoholic fatty liver disease. Hepatology 42: 880–885.
[19]
La Cava A, Van Kaer L, Fu Dong S (2006) CD4+CD25+ Tregs and NKT cells: regulators regulating regulators. Trends Immunol 27: 322–327.
[20]
Jiang S, Game DS, Davies D, Lombardi G, Lechler RI (2005) Activated CD1d-restricted natural killer T cells secrete IL-2: innate help for CD4+CD25+ regulatory T cells? Eur J Immunol 35: 1193–1200.
[21]
Pillai AB, George TI, Dutt S, Strober S (2009) Host natural killer T cells induce an interleukin-4-dependent expansion of donor CD4+CD25+Foxp3+ T regulatory cells that protects against graft-versus-host disease. Blood 113: 4458–4467.
[22]
Santodomingo-Garzon T, Han J, Le T, Yang Y, Swain MG (2009) Natural killer T cells regulate the homing of chemokine CXC receptor 3-positive regulatory T cells to the liver in mice. Hepatology 49: 1267–1276.
[23]
Azuma T, Takahashi T, Kunisato A, Kitamura T, Hirai H (2003) Human CD4+ CD25+ Regulatory T Cells Suppress NKT Cell Functions. Cancer Res 63: 4516–4520.
[24]
Hallett WH, Ames E, Alvarez M, Barao I, Taylor PA, et al. (2008) Combination therapy using IL-2 and anti-CD25 results in augmented natural killer cell-mediated antitumor responses. Biol Blood Marrow Transplant 14: 1088–1099.
[25]
Tupin E, Kronenberg M (2006) Activation of natural killer T cells by glycolipids. Methods Enzymol 417: 185–201.
[26]
Zinchuk V, Zinchuk O (2008) Quantitative colocalization analysis of confocal fluorescence microscopy images. Curr Protoc Cell Biol Chapter 4: Unit 4 19:
[27]
Yang SQ, Lin HZ, Diehl AM (2001) Fatty liver vulnerability to lipopolysaccaride despite NF-kB induction and caspase 3 inhibition. Am J Physiol 281: G382–G392.
[28]
Chomczynski P, Sacchi N (1987) Single step method of RNA isolation by acid guanidine thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–161.
[29]
Bergmeyer HU, Scheibe P, Wahlefeld AW (1978) Optimization of methods for aspartate aminotransferase and alanine aminotransferase. Clin Chem 24: 58–73.
[30]
Li W, Ji F, Zhang Y, Wang Y, Yang N, et al. (2008) Cooperation of invariant NKT cells and CD4+CD25+ T regulatory cells in prevention of autoimmune diabetes in non-obese diabetic mice treated with alpha-galactosylceramide. Acta Biochim Biophys Sin (Shanghai) 40: 381–390.
[31]
Vela-Ojeda J, Montiel-Cervantes L, Granados-Lara P, Reyes-Maldonado E, Garcia-Latorre E, et al. (2010) Role of CD4+CD25+(high)Foxp3+CD62L+ Regulatory T Cells and Invariant NKT Cells in Human Allogeneic Hematopoietic Stem Cell Transplantation. Stem Cells Dev.
[32]
Nador RG, Hongo D, Yao Z, Strober S (2010) The Changed Balance of Regulatory and Naive T Cells Promotes Tolerance after TLI and Anti-T-Cell Antibody Conditioning. American Journal of Transplantation 10: 262–272.
[33]
Grossman WJ, Verbsky JW, Barchet W, Colonna M, Atkinson JP, et al. (2004) Human T regulatory cells can use the perforin pathway to cause autologous target cell death. Immunity 21: 589–601.
[34]
Takeda K, Hayakawa Y, Van-Kaer L, Matsuda H, Yagita H, et al. (2000) Critical contribution of liver natural killer T cells to a murine model of hepatitis. Proc Natl Acad Sci U S A 97: 5498–5503.
[35]
Yasser O, Toshihiko K, Tetsuya N, Kazuyoshi T, Luc？Van K, et al. (2000) Activation of hepatic NKT cells and subsequent liver injury following administration of α-galactosylceramide. European Journal of Immunology 30: 1919–1928.
[36]
Ma X, Hua J, Mohamood AR, Hamad AR, Ravi R, et al. (2007) A high-fat diet and regulatory T cells influence susceptibility to endotoxin-induced liver injury. Hepatology 46: 1519–1529.
[37]
Bendelac A, Savage PB, Teyton L (2007) The biology of NKT cells. Annu Rev Immunol 25: 297–336.
[38]
Godfrey DI, Kronenberg M (2004) Going both ways: Immune regulation via CD1d-dependent NKT cells. J Clin Invest 114: 1379–1388.
[39]
Coquet JM, Chakravarti S, Kyparissoudis K, McNab FW, Pitt LA, et al. (2008) Diverse cytokine production by NKT cell subsets and identification of an IL-17-producing CD4-NK1.1- NKT cell population. Proc Natl Acad Sci U S A 105: 11287–11292.
[40]
Sakaguchi S (2004) Naturally arising CD4+ regulatory t cells for immunologic self-tolerance and negative control of immune responses. Annu Rev Immunol 22: 531–562.
[41]
Longhi MS, Hussain MJ, Mitry RR, Arora SK, Mieli-Vergani G, et al. (2006) Functional study of CD4+CD25+ regulatory T cells in health and autoimmune hepatitis. J Immunol 176: 4484–4491.
[42]
Kita H, Naidenko OV, Kronenberg M, Ansari AA, Rogers P, et al. (2002) Quantitation and phenotypic analysis of natural killer T cells in primary biliary cirrhosis using a human CD1d tetramer. Gastroenterology 123: 1031–1043.
[43]
Onishi Y, Fehervari Z, Yamaguchi T, Sakaguchi S (2008) Foxp3+ natural regulatory T cells preferentially form aggregates on dendritic cells in vitro and actively inhibit their maturation. Proc Natl Acad Sci U S A 105: 10113–10118.
