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.

  • Article
  • Published:

SRSF1 plays a critical role in invariant natural killer T cell development and function

Cellular & Molecular Immunology volume 18, pages 2502–2515 (2021)Cite this article

Subjects

Abstract

Invariant natural killer T (iNKT) cells are highly conserved innate-like T lymphocytes that originate from CD4+CD8+ double-positive (DP) thymocytes. Here, we report that serine/arginine splicing factor 1 (SRSF1) intrinsically regulates iNKT cell development by directly targeting Myb and balancing the abundance of short and long isoforms. Conditional ablation of SRSF1 in DP cells led to a substantially diminished iNKT cell pool due to defects in proliferation, survival, and TCRα rearrangement. The transition from stage 0 to stage 1 of iNKT cells was substantially blocked, and the iNKT2 subset was notably diminished in SRSF1-deficient mice. SRSF1 deficiency resulted in aberrant expression of a series of regulators that are tightly correlated with iNKT cell development and iNKT2 differentiation, including Myb, PLZF, Gata3, ICOS, and CD5. In particular, we found that SRSF1 directly binds and regulates pre-mRNA alternative splicing of Myb and that the expression of the short isoform of Myb is substantially reduced in SRSF1-deficient DP and iNKT cells. Strikingly, ectopic expression of the Myb short isoform partially rectified the defects caused by ablation of SRSF1. Furthermore, we confirmed that the SRSF1-deficient mice exhibited resistance to acute liver injury upon α-GalCer and Con A induction. Our findings thus uncovered a previously unknown role of SRSF1 as an essential post-transcriptional regulator in iNKT cell development and functional differentiation, providing new clinical insights into iNKT-correlated disease.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Pellicci DG, Koay HF, Berzins SP. Thymic development of unconventional T cells: how NKT cells, MAIT cells and gammadelta T cells emerge. Nat Rev Immunol. 2020;12:756–70.

    Article  CAS  Google Scholar 

  2. Lantz O, Bendelac A. An invariant T cell receptor alpha chain is used by a unique subset of major histocompatibility complex class I-specific CD4+ and CD4-8- T cells in mice and humans. J Exp Med. 1994;180:1097–106.

    Article  CAS  PubMed  Google Scholar 

  3. Adachi Y, Koseki H, Zijlstra M, Taniguchi M. Positive selection of invariant V alpha 14+ T cells by non-major histocompatibility complex-encoded class I-like molecules expressed on bone marrow-derived cells. Proc Natl Acad Sci USA. 1995;92:1200–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kawano T, Cui J, Koezuka Y, Toura I, Kaneko Y, Motoki K, et al. CD1d-restricted and TCR-mediated activation of valpha14 NKT cells by glycosylceramides. Science. 1997;278:1626–9.

    Article  CAS  PubMed  Google Scholar 

  5. Beckman EM, Porcelli SA, Morita CT, Behar SM, Furlong ST, Brenner MB. Recognition of a lipid antigen by CD1-restricted alpha beta+ T cells. Nature 1994;372:691–4.

    Article  CAS  PubMed  Google Scholar 

  6. Bendelac A, Savage PB, Teyton L. The biology of NKT cells. Annu Rev Immunol. 2007;25:297–336.

    Article  CAS  PubMed  Google Scholar 

  7. Godfrey DI, Berzins SP. Control points in NKT-cell development. Nat Rev Immunol. 2007;7:505–18.

    Article  CAS  PubMed  Google Scholar 

  8. Lee YJ, Holzapfel KL, Zhu J, Jameson SC, Hogquist KA. Steady-state production of IL-4 modulates immunity in mouse strains and is determined by lineage diversity of iNKT cells. Nat Immunol. 2013;14:1146–54.

    Article  CAS  PubMed  Google Scholar 

  9. Harsha Krovi S, Zhang J, Michaels-Foster MJ, Brunetti T, Loh L, Scott-Browne J, et al. Thymic iNKT single cell analyses unmask the common developmental program of mouse innate T cells. Nat Commun. 2020;11:6238.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Baranek T, Lebrigand K, de Amat Herbozo C, Gonzalez L, Bogard G, Dietrich C, et al. High dimensional single-cell analysis reveals iNKT cell developmental trajectories and effector fate decision. Cell Rep. 2020;32:108116.

    Article  CAS  PubMed  Google Scholar 

  11. Coquet JM, Chakravarti S, Kyparissoudis K, McNab FW, Pitt LA, McKenzie BS, et al. Diverse cytokine production by NKT cell subsets and identification of an IL-17-producing CD4-NK1.1- NKT cell population. Proc Natl Acad Sci USA. 2008;105:11287–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Brennan PJ, Brigl M, Brenner MB. Invariant natural killer T cells: an innate activation scheme linked to diverse effector functions. Nat Rev Immunol. 2013;13:101–17.

    Article  CAS  PubMed  Google Scholar 

  13. Crosby CM, Kronenberg M. Tissue-specific functions of invariant natural killer T cells. Nat Rev Immunol. 2018;18:559–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Dong Z, Wei H, Sun R, Tian Z. The roles of innate immune cells in liver injury and regeneration. Cell Mol Immunol. 2007;4:241–52.

    CAS  PubMed  Google Scholar 

  15. Wang H, Feng D, Park O, Yin S, Gao B. Invariant NKT cell activation induces neutrophil accumulation and hepatitis: opposite regulation by IL-4 and IFN-gamma. Hepatology 2013;58:1474–85.

    Article  CAS  PubMed  Google Scholar 

  16. Heymann F, Hamesch K, Weiskirchen R, Tacke F. The concanavalin A model of acute hepatitis in mice. Lab Anim. 2015;49(1 Suppl):12–20.

    Article  CAS  PubMed  Google Scholar 

  17. Liew PX, Lee WY, Kubes P. iNKT cells orchestrate a switch from inflammation to resolution of sterile liver injury. Immunity 2017;47:752–65 e5.

    Article  CAS  PubMed  Google Scholar 

  18. Shissler SC, Webb TJ. The ins and outs of type I iNKT cell development. Mol Immunol. 2019;105:116–30.

    Article  CAS  PubMed  Google Scholar 

  19. Godfrey DI, Stankovic S, Baxter AG. Raising the NKT cell family. Nat Immunol. 2010;11:197–206.

    Article  CAS  PubMed  Google Scholar 

  20. Gapin L. Development of invariant natural killer T cells. Curr Opin Immunol. 2016;39:68–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kovalovsky D, Uche OU, Eladad S, Hobbs RM, Yi W, Alonzo E, et al. The BTB-zinc finger transcriptional regulator PLZF controls the development of invariant natural killer T cell effector functions. Nat Immunol. 2008;9:1055–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Savage AK, Constantinides MG, Han J, Picard D, Martin E, Li B, et al. The transcription factor PLZF directs the effector program of the NKT cell lineage. Immunity 2008;29:391–403.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Hu T, Simmons A, Yuan J, Bender TP, Alberola-Ila J. The transcription factor c-Myb primes CD4+CD8+ immature thymocytes for selection into the iNKT lineage. Nat Immunol. 2010;11:435–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Wikenheiser DJ, Stumhofer JS. ICOS co-stimulation: friend or foe? Front Immunol. 2016;7:304.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Akbari O, Stock P, Meyer EH, Freeman GJ, Sharpe AH, Umetsu DT, et al. ICOS/ICOSL interaction is required for CD4+ invariant NKT cell function and homeostatic survival. J Immunol. 2008;180:5448–56.

    Article  CAS  PubMed  Google Scholar 

  26. Kim PJ, Pai SY, Brigl M, Besra GS, Gumperz J, Ho IC. GATA-3 regulates the development and function of invariant NKT cells. J Immunol. 2006;177:6650–9.

    Article  CAS  PubMed  Google Scholar 

  27. Seiler MP, Mathew R, Liszewski MK, Spooner CJ, Barr K, Meng F, et al. Elevated and sustained expression of the transcription factors Egr1 and Egr2 controls NKT lineage differentiation in response to TCR signaling. Nat Immunol. 2012;13:264–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Townsend MJ, Weinmann AS, Matsuda JL, Salomon R, Farnham PJ, Biron CA, et al. T-bet regulates the terminal maturation and homeostasis of NK and Valpha14i NKT cells. Immunity 2004;20:477–94.

    Article  CAS  PubMed  Google Scholar 

  29. Weinreich MA, Odumade OA, Jameson SC, Hogquist KA. T cells expressing the transcription factor PLZF regulate the development of memory-like CD8+ T cells. Nat Immunol. 2010;11:709–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Li J, Wu D, Jiang N, Zhuang Y. Combined deletion of Id2 and Id3 genes reveals multiple roles for E proteins in invariant NKT cell development and expansion. J Immunol. 2013;191:5052–64.

    Article  CAS  PubMed  Google Scholar 

  31. Li J, Roy S, Kim YM, Li S, Zhang B, Love C, et al. Id2 collaborates with Id3 To suppress invariant NKT and innate-like tumors. J Immunol. 2017;198:3136–48.

    Article  CAS  PubMed  Google Scholar 

  32. Dvinge H, Kim E, Abdel-Wahab O, Bradley RK. RNA splicing factors as oncoproteins and tumour suppressors. Nat Rev Cancer. 2016;16:413–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Jeong SSR. Proteins: binders, regulators, and connectors of RNA. Mol Cells. 2017;40:1–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Das S, Krainer AR. Emerging functions of SRSF1, splicing factor and oncoprotein, in RNA metabolism and cancer. Mol Cancer Res. 2014;12:1195–204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Xu X, Yang D, Ding JH, Wang W, Chu PH, Dalton ND, et al. ASF/SF2-regulated CaMKIIdelta alternative splicing temporally reprograms excitation-contraction coupling in cardiac muscle. Cell 2005;120:59–72.

    Article  CAS  PubMed  Google Scholar 

  36. Hennet T, Hagen FK, Tabak LA, Marth JD. T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination. Proc Natl Acad Sci USA. 1995;92:12070–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Liu J, Cui Z, Wang F, Yao Y, Yu G, Liu J, et al. Lrp5 and Lrp6 are required for maintaining self-renewal and differentiation of hematopoietic stem cells. FASEB J. 2019;33:5615–25.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Kim TJ, Park G, Kim J, Lim SA, Kim J, Im K, et al. CD160 serves as a negative regulator of NKT cells in acute hepatic injury. Nat Commun. 2019;10:3258.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Yao Y, Guo W, Chen J, Guo P, Yu G, Liu J, et al. Long noncoding RNA Malat1 is not essential for T cell development and response to LCMV infection. RNA Biol. 2018;15:1477–86.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Bedel R, Berry R, Mallevaey T, Matsuda JL, Zhang J, Godfrey DI, et al. Effective functional maturation of invariant natural killer T cells is constrained by negative selection and T-cell antigen receptor affinity. Proc Natl Acad Sci USA. 2014;111:E119–28.

    Article  CAS  PubMed  Google Scholar 

  41. Yu S, Zhou X, Steinke FC, Liu C, Chen SC, Zagorodna O, et al. The TCF-1 and LEF-1 transcription factors have cooperative and opposing roles in T cell development and malignancy. Immunity 2012;37:813–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Godfrey DI, MacDonald HR, Kronenberg M, Smyth MJ, Van Kaer L. NKT cells: what’s in a name? Nat Rev Immunol. 2004;4:231–7.

    Article  CAS  PubMed  Google Scholar 

  43. Guo J, Hawwari A, Li H, Sun Z, Mahanta SK, Littman DR, et al. Regulation of the TCRalpha repertoire by the survival window of CD4(+)CD8(+) thymocytes. Nat Immunol. 2002;3:469–76.

    Article  PubMed  CAS  Google Scholar 

  44. Das R, Sant’Angelo DB, Nichols KE. Transcriptional control of invariant NKT cell development. Immunol Rev. 2010;238:195–215.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Baker SJ, Kumar A, Reddy EP. p89c-Myb is not required for fetal or adult hematopoiesis. Genesis 2010;48:309–16.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Zhu S, Zhang H, Bai L. NKT cells in liver diseases. Front Med. 2018;12:249–61.

    Article  PubMed  Google Scholar 

  47. Bandyopadhyay K, Marrero I, Kumar V. NKT cell subsets as key participants in liver physiology and pathology. Cell Mol Immunol. 2016;13:337–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Huang W, He W, Shi X, He X, Dou L, Gao Y. The role of CD1d and MR1 restricted T cells in the liver. Front Immunol. 2018;9:2424.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Dennert G, Aswad F. The role of NKT cells in animal models of autoimmune hepatitis. Crit Rev Immunol. 2006;26:453–73.

    Article  CAS  PubMed  Google Scholar 

  50. Anczukow O, Rosenberg AZ, Akerman M, Das S, Zhan L, Karni R, et al. The splicing factor SRSF1 regulates apoptosis and proliferation to promote mammary epithelial cell transformation. Nat Struct Mol Biol. 2012;19:220–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Zhou X, Wang R, Li X, Yu L, Hua D, Sun C, et al. Splicing factor SRSF1 promotes gliomagenesis via oncogenic splice-switching of MYO1B. J Clin Invest. 2019;129:676–93.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Katsuyama T, Martin-Delgado IJ, Krishfield SM, Kyttaris VC, Moulton VR. Splicing factor SRSF1 controls T cell homeostasis and its decreased levels are linked to lymphopenia in systemic lupus erythematosus. Rheumatology. 2020;59:2146–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Li X, Wang J, Manley JL. Loss of splicing factor ASF/SF2 induces G2 cell cycle arrest and apoptosis, but inhibits internucleosomal DNA fragmentation. Genes Dev. 2005;19:2705–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Nakata Y, Brignier AC, Jin S, Shen Y, Rudnick SI, Sugita M, et al. c-Myb, Menin, GATA-3, and MLL form a dynamic transcription complex that plays a pivotal role in human T helper type 2 cell development. Blood 2010;116:1280–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Rooney S, Sekiguchi J, Zhu C, Cheng HL, Manis J, Whitlow S, et al. Leaky Scid phenotype associated with defective V(D)J coding end processing in Artemis-deficient mice. Mol Cell. 2002;10:1379–90.

    Article  CAS  PubMed  Google Scholar 

  56. Qi Z, Wang F, Yu G, Wang D, Yao Y, You M, et al. SRSF1 serves as a critical posttranscriptional regulator at the late stage of thymocyte development. Sci Adv. 2021;16:eabf0753.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by grants from the National Key Research and Development Program of China (2017YFA0104401), the National Natural Scientific Foundation of China (32130039, 31970831, and 31630038), and the Project for Extramural Scientists of State Key Laboratory of Agrobiotechnology from China Agricultural University (2021SKLAB6-3, 2021SKLAB6-4, 2019SKLAB6-6, and 2019SKLAB6-7).

Author information

Author notes

  1. These authors contributed equally: Jingjing Liu, Menghao You.

Authors and Affiliations

  1. State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China

    Jingjing Liu, Menghao You, Yingpeng Yao, Ce Ji, Zhao Wang, Fang Wang, Zhihong Qi, Guotao Yu, Zhen Sun, Wenhui Guo, Juanjuan Liu, Tianyan Zhao & Shuyang Yu

  2. Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

    Di Wang & Yuanchao Xue

  3. Clinical Department, College of Veterinary Medicine, China Agricultural University, Beijing, China

    Shumin Li & Yipeng Jin

  4. Center for Discovery and Innovation, Hackensack Meridian Health, Nutley, NJ, USA

    Hai-Hui Xue

Authors
  1. Jingjing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Menghao You
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yingpeng Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ce Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Di Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhihong Qi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Guotao Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Zhen Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Wenhui Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Juanjuan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Shumin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Yipeng Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Tianyan Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Hai-Hui Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Yuanchao Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Shuyang Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Jingjing L., M.Y., and C.J. performed the major experiments and analyzed the data; M.Y., Z.W., and F.W. analyzed the high-throughput data; D.W., Z.Q., Y.Y., G.Y., Z.S., W.G., Juanjuan L., S.L., Y.J., and T.Z. assisted with the overall experiments; S.Y. designed and supervised the experiments with constructive suggestions from H.-H.X. and Y.X.; Y.Y., Jingjing L., and S.Y. wrote the manuscript with revisions from all authors.

Corresponding author

Correspondence to Shuyang Yu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, J., You, M., Yao, Y. et al. SRSF1 plays a critical role in invariant natural killer T cell development and function. Cell Mol Immunol 18, 2502–2515 (2021). https://doi.org/10.1038/s41423-021-00766-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41423-021-00766-w

Keywords

This article is cited by

Search

Advanced search

Quick links