Proliferation and Directional Differentiation of iNKT Cells Derived from DBA/1 Mice Thymus
-
Dongzhi Chen
,
Zhao Li
,
Rui Liang
,
Huifang Liu
,
Yuanyuan Wang
,
Hongyun Shi
,
Ming Meng
Abstract
The rates of invariant natural killer T (iNKT) cells in vivo are very low, and the amounts of cells obtained directly from the body are hard enough to fulfill their potential in clinical application.
To overcome this problem, we subcutaneously injected alpha-galactosylceramide (α-GalCer) into DBA/1 mice and thymic single cells were isolated and cultured in vitro. Fluorescence-activated cell sorting was used to detect the iNKT cells and their subsets in the thymus after the injection of α-GalCer by different methods. In addition, in vitro changes of single-cell suspensions and their cytokines in culture supernatants were assessed.
Compared with the α-GalCer multiple subcutaneous injection group, the rates of iNKT cells in the α-GalCer single subcutaneous injection group were markedly higher at each time point, while the highest levels of iNKT1 and iNKT2 cells were observed on day 4 and 8, respectively. In α-GalCer single subcutaneous injection for 8 days and thymic mononuclear cell cultured for 14 days group, the expansion rate of iNKT cells was significantly faster than the other groups, while it reached a peak for iNKT1 cells. Interferon-gamma was consistent with the development of iNKT1 cells, however no difference was found between the cultured iNKT cells in vitro and the natural iNKT cells in vivo in terms of cytokine production.
Herein, we introduced a method in which antigenic stimulation in vivo and directed induction in vitro yielded high levels of iNKT cells with specific functions.
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17. Cohen NR, Garg S, Brenner MB. Antigen presentation by CD1 lipids, T cells, and NKT cells in microbial immunity. Adv Immunol. 2009;102(8):1–94.
18. Kohlgruber AC, Donado CA, LaMarche NM, Brenner MB, Brennan PJ. Activation strategies for invariant natural killer T cells. Immunogenetics. 2016;68(12):649–63.
19. Kim E Y, Lynch L, Brennan PJ, Cohen NR, Brenner MB. The transcriptional programs of iNKT cells. Semin Immunol.2015;27(4):26–32.
20. Kumar A, Suryadevara N, Hill TM, Bezbradica JS, Van Kaer L, Joyce S. Natural Killer T Cells: An Ecological Evolutionary Developmental Biology Perspective. Front Immunol. 2017;8:1858.
21. Crosby CM, Kronenberg M. Tissue-specific functions of invariant natural killer T cells.Nat Rev Immunol. 2018;18(9):559-74.
22. Van Kaer, Wu L. Therapeutic Potential of Invariant Natural Killer T Cells in Autoimmunity. Front Immunol.2018;9:519.
23. Govindarajan S, Elewaut D, Drennan M. An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen. J Vis Exp. 2015;29(105):e53256.
24. Tuttle KD, Gapin L. Characterization of Thymic Development of Natural Killer T Cell Subsets by Multiparameter Flow Cytometry. Methods Mol Biol. 2018;1799(114);121-33.
25. Carr T, Krishnamoorthy V, Yu S, Xue HH, Kee BL, Verykokakis M. The transcription factor lymphoid enhancer factor 1 controls invariant natural killer T cell expansion and Th2-type effector differentiation. J Exp Med. 2015;212(5):793-807.
26. Das R, Sant’Angelo DB, Nichols KE. Transcriptional control of invariant NKT cell development. Immunol Rev. 2010;238(12):195-215.
27. Wong C H, Jenne C N, Lee W Y, Léger C, Kubes P. Functional innervation of hepatic iNKT cells is immunosuppressive following stroke. Science. 2011;334(42):101-5.
28. Nieuwenhuis EE, Matsumoto T, Lindenbergh D, Willemsen R, Kaser A, Simons-Oosterhuis Y, et al. Cd1d-dependent regulation of bacterial colonization in the intestine of mice. J ClinInvest. 2009;119(14):1241-50.
29. Paget C, Ivanov S, Fontaine J, Renneson J, Blanc F, Pichavant M, Dumoutier L, et al. Interleukin-22 is produced by invariant natural killer T lymphocytes during influenza A virus infection: potential role in protection against lung epithelial damages. J Biol Chem. 2012; 287(11):8816-29.
30. De Santo C, Salio M, Masri SH, Lee LY, Dong T, Speak AO, et al. Invariant NKT cells reduce the immunosuppressive activity of influenza A virus-induced myeloid-derived suppressor cells in mice and humans. J Clin Invest. 2008;118(21):4036–48.
31. Matsuda JL, Mallevaey T, Scott-Browne J, Gapin L. CD1d-restricted iNKT cells, the Swiss-Army knife' of the immune system. Curr Opin Immunol. 2008; 20(2):358-68.
32. Parekh VV, Wilson MT, Olivares-Villagómez D, Singh AK, Wu L, Wang CR, et al. Glycolipid antigen induces long-term natural killer T cell anergy in mice. J Clin Invest. 2005;115(9):2572-83.
33. Sharma A1, Berga-Bolanos R, Sultana DA, Sen JM. IL-4 and IL-4 Receptor Expression Is Dispensable for the Development and Function of Natural Killer T Cells. Plos One. 2013;(8):e71872.
34. Iizuka A, Ikarashi Y, Yoshida M, Heike Y, Takeda K, Quinn G, Wakasugi H, et al.Interleukin (IL)-4 promotes T helper type 2-biased natural killer T (NKT) cell expansion, which is regulated by NKT cell-derived interferon-gamma and IL-4. Insect Science. 2010;123(1):100-7.
35. Liu XF, Wu YH, Wei SN, Wang N, Li PF, Li YZ, et al. Establishment and characterization of a kidney cell line from kelp grouper Epinephelusmoara. Fish PhysiolBiochem. 2018;44(1):87-93.
36. Yao X, Zhao J, Kong Q, Xie X, Wang J, Sun B, et al. Exogenous IL-9 Ameliorates Experimental Autoimmune Myasthenia Gravis Symptoms in Rats. Immunol. Invest.2018;47(7):712-24.
37. Chiba A1, Cohen N, Brigl M, Brennan PJ, Besra GS, Brenner MB. Rapid and reliable generation of invariant natural killer T-cell lines in vitro. Immunology. 2010;128(3):324-33.
38. Molling JW, Moreno M, van der Vliet HJ, von Blomberg BM, van den Eertwegh AJ, Scheper RJ,et al. Generation and sustained expansion of mouse spleen invariant NKT cell lines with preserved cytokine releasing capacity. J Immunol Methods. 2007;322(1-2):70-81.
2. Zhu S, Zhang H, Bai L. NKT cells in liver diseases. Frontiers of Medicine, (7):1-13,2018
3. Krijgsman D, Hokland M, Kuppen PJK. The Role of Natural Killer T Cells in Cancer-A Phenotypical and Functional Approach. Front Immunol. 2018;9:367-9.
4. Gumperz JE. The Ins and Outs of CD1 Molecules: Bringing Lipids Under Immunological Surveillance. Traffic. 2006;7(1):2-13.
5. Clancy-Thompson E, Chen GZ, Tyler PM. Monoclonal Invariant NKT (iNKT) Cell Mice Reveal a Role for Both Tissue of Origin and the TCR in Development of iNKT Functional Subsets. J Immunol. 2017;199(11):159-71.
6. Pellicci DG, Uldrich AP. Unappreciated diversity within the pool of CD1d-restricted T cells. Semin Cell Dev Biol. 2018;84(4):42-7.
7. Bennstein SB. Unraveling Natural Killer T-Cells Development. Front Immunol. 2018;8(1):1950-9.
8. Lee Y J, Wang H, Starrett G J, Phuong V, Jameson SC, Hogquist KA. Tissue-Specific Distribution of iNKT Cells Impacts Their Cytokine Response. Immunity.2015;43(5):566-78.
9. White A J, Lucas B, Jenkinson WE, Anderson G. Invariant NKT Cells and Control of the Thymus Medulla. J Immunol. 2018;200:333-9.
10. Cameron G and Godfrey DI: Differential surface phenotype and context-dependent reactivity of functionally diverse NKT cells. Immunol Cell Biol.2018;10:1111.
11. Pellicci DG, Hammond KJ, Uldrich AP, Baxter AG, Smyth MJ, Godfrey DI. A natural killer T (NKT) cell developmental pathway involving a thymus-dependent NK1.1(-) CD4(+) CD1d-dependent precursor stage. J Exp Med. 2002;7:835–44.
12. LynchL, MicheletX, ZhangS. Regulatory iNKT cells lack expression of the transcription factor PLZF and control the homeostasis of T(reg) cells and macrophages in adipose tissue [J]. Nat Immunol. 2015;16(1):85–95.
13. Gao B, Jeong WI, Tian Z.Liver: An organ with predominant innate immunity. Hepatology. 2010;47(2):729-36.
14. Gray EE, Friend S, Suzuki K, Phan TG, Cyster JG. Subcapsular sinus macrophage fragmentation and CD169+ bleb acquisition by closely associated IL-17-committed innate-like lymphocytes. PLoS One. 2012;7(6):e38258.
15. Kwon DI, Lee YJ. Lineage Differentiation Program of Invariant Natural Killer T Cells. Immune Netw. 2017;17(6):365-77.
16. Brennan P J, Brigl M, Brenner MB. Invariant natural killer T cells: an innate activation scheme linked to diverse effector functions. Nat Rev Immunol.2013;13(8):101–17.
17. Cohen NR, Garg S, Brenner MB. Antigen presentation by CD1 lipids, T cells, and NKT cells in microbial immunity. Adv Immunol. 2009;102(8):1–94.
18. Kohlgruber AC, Donado CA, LaMarche NM, Brenner MB, Brennan PJ. Activation strategies for invariant natural killer T cells. Immunogenetics. 2016;68(12):649–63.
19. Kim E Y, Lynch L, Brennan PJ, Cohen NR, Brenner MB. The transcriptional programs of iNKT cells. Semin Immunol.2015;27(4):26–32.
20. Kumar A, Suryadevara N, Hill TM, Bezbradica JS, Van Kaer L, Joyce S. Natural Killer T Cells: An Ecological Evolutionary Developmental Biology Perspective. Front Immunol. 2017;8:1858.
21. Crosby CM, Kronenberg M. Tissue-specific functions of invariant natural killer T cells.Nat Rev Immunol. 2018;18(9):559-74.
22. Van Kaer, Wu L. Therapeutic Potential of Invariant Natural Killer T Cells in Autoimmunity. Front Immunol.2018;9:519.
23. Govindarajan S, Elewaut D, Drennan M. An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen. J Vis Exp. 2015;29(105):e53256.
24. Tuttle KD, Gapin L. Characterization of Thymic Development of Natural Killer T Cell Subsets by Multiparameter Flow Cytometry. Methods Mol Biol. 2018;1799(114);121-33.
25. Carr T, Krishnamoorthy V, Yu S, Xue HH, Kee BL, Verykokakis M. The transcription factor lymphoid enhancer factor 1 controls invariant natural killer T cell expansion and Th2-type effector differentiation. J Exp Med. 2015;212(5):793-807.
26. Das R, Sant’Angelo DB, Nichols KE. Transcriptional control of invariant NKT cell development. Immunol Rev. 2010;238(12):195-215.
27. Wong C H, Jenne C N, Lee W Y, Léger C, Kubes P. Functional innervation of hepatic iNKT cells is immunosuppressive following stroke. Science. 2011;334(42):101-5.
28. Nieuwenhuis EE, Matsumoto T, Lindenbergh D, Willemsen R, Kaser A, Simons-Oosterhuis Y, et al. Cd1d-dependent regulation of bacterial colonization in the intestine of mice. J ClinInvest. 2009;119(14):1241-50.
29. Paget C, Ivanov S, Fontaine J, Renneson J, Blanc F, Pichavant M, Dumoutier L, et al. Interleukin-22 is produced by invariant natural killer T lymphocytes during influenza A virus infection: potential role in protection against lung epithelial damages. J Biol Chem. 2012; 287(11):8816-29.
30. De Santo C, Salio M, Masri SH, Lee LY, Dong T, Speak AO, et al. Invariant NKT cells reduce the immunosuppressive activity of influenza A virus-induced myeloid-derived suppressor cells in mice and humans. J Clin Invest. 2008;118(21):4036–48.
31. Matsuda JL, Mallevaey T, Scott-Browne J, Gapin L. CD1d-restricted iNKT cells, the Swiss-Army knife' of the immune system. Curr Opin Immunol. 2008; 20(2):358-68.
32. Parekh VV, Wilson MT, Olivares-Villagómez D, Singh AK, Wu L, Wang CR, et al. Glycolipid antigen induces long-term natural killer T cell anergy in mice. J Clin Invest. 2005;115(9):2572-83.
33. Sharma A1, Berga-Bolanos R, Sultana DA, Sen JM. IL-4 and IL-4 Receptor Expression Is Dispensable for the Development and Function of Natural Killer T Cells. Plos One. 2013;(8):e71872.
34. Iizuka A, Ikarashi Y, Yoshida M, Heike Y, Takeda K, Quinn G, Wakasugi H, et al.Interleukin (IL)-4 promotes T helper type 2-biased natural killer T (NKT) cell expansion, which is regulated by NKT cell-derived interferon-gamma and IL-4. Insect Science. 2010;123(1):100-7.
35. Liu XF, Wu YH, Wei SN, Wang N, Li PF, Li YZ, et al. Establishment and characterization of a kidney cell line from kelp grouper Epinephelusmoara. Fish PhysiolBiochem. 2018;44(1):87-93.
36. Yao X, Zhao J, Kong Q, Xie X, Wang J, Sun B, et al. Exogenous IL-9 Ameliorates Experimental Autoimmune Myasthenia Gravis Symptoms in Rats. Immunol. Invest.2018;47(7):712-24.
37. Chiba A1, Cohen N, Brigl M, Brennan PJ, Besra GS, Brenner MB. Rapid and reliable generation of invariant natural killer T-cell lines in vitro. Immunology. 2010;128(3):324-33.
38. Molling JW, Moreno M, van der Vliet HJ, von Blomberg BM, van den Eertwegh AJ, Scheper RJ,et al. Generation and sustained expansion of mouse spleen invariant NKT cell lines with preserved cytokine releasing capacity. J Immunol Methods. 2007;322(1-2):70-81.
| Files | ||
| Issue | Vol 20 No 5 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v20i5.7412 | |
| Keywords | ||
| Alpha-galactosylceramide Cell differentiation Thymus gland | ||
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How to Cite
1.
Chen D, Li Z, Liang R, Liu H, Wang Y, Shi H, Meng M. Proliferation and Directional Differentiation of iNKT Cells Derived from DBA/1 Mice Thymus. Iran J Allergy Asthma Immunol. 2021;20(5):623-634.
