Advertisement

Biotechnology Letters

, Volume 41, Issue 10, pp 1095–1104 | Cite as

Towards a surrogate system to express human lipid binding TCRs

  • Rui Wang
  • Ronja Pscheid
  • Ashfaq Ghumra
  • Ling Yu Lea Kan
  • Stella Cochrane
  • Lucy Fairclough
  • Marcos J. C. AlcocerEmail author
Original Research Paper

Abstract

Background

Previously we reported that natural nut lipids were necessary for sensitization and that natural killer T cells (NKTs) must play a critical role in the development of food allergic responses. A major bottleneck in further understanding the interaction of nut lipids with the cells of the human immune system is the lack of well-characterized lipid responsive human cell lines.

Objective

In the present study, we engineered human T cell receptor (TCR) sequences TRAV10 and TRBV25 responsive to α-GalCer into a stable murine iNKT hybridoma and surrogate human T cell lines.

Results

The murine hybridoma system has shown to be problematic. To overcome this limitation, the expression of human TCR α/β sequences has been achieved driven by a bidirectional promoter on a plasmids or a lentivirus system, employing stable DC cell lines as lipid presenting cells, and a stable T cell line as a surrogate system. Further, a commercial human Jurkat T cell line containing an inducible secreted luciferase reporter construct was shown to be functional and can be used for a transient expression of human TCRs in a lipid screening program. The transfection efficiencies were improved using the lentivirus polycistronic constructs containing the P2A sequence in a TCR αβ/γδ null cell line (Jurkat 76).

Conclusions

The results suggest that the mis-pairing of the endogenous α/β TCR during ER folding in the presence of the new human TCR sequences could be impairing the functionality of the TCR lipid receptors. The surrogate systems presented here are important first steps in the establishment of human cell-specific lipid responsive libraries for the study of natural lipid substances.

Keywords

Food allergy NKT TCR Lipid binding TCR Dendritic cells Mutz JawsII In vitro system 

Notes

Acknowledgements

We would like also to thank Dr Cinzia Allegrucci, School of Veterinary, University of Nottingham for support on the lentivirus work. A.F. has been financially supported by a Research Grant (534231) from Unilever UK Central resources, hence the execution of the work has been partially funded and the final article for publication reviewed by Unilever.

Funding

This study was partially funded by a Research Grant (534231) from Unilever UK Central resources.

Compliance with ethical standards

Conflict of interest

Author A. Ghumra received research grant from Unilever UK Central resources, hence the execution of the work has been partially funded and the final article for publication reviewed by Unilever.

References

  1. Ahmadi M, King JW, Xue SA et al (2011) CD3 limits the efficacy of TCR gene therapy in vivo. Blood 118:3528–3537.  https://doi.org/10.1182/blood-2011-04-346338 CrossRefGoogle Scholar
  2. Bai L, Constantinides MG, Thomas SY et al (2012) Distinct APCs explain the cytokine bias of α-galactosylceramide variants in vivo. J Immunol 188:3053–3061.  https://doi.org/10.4049/jimmunol.1102414 CrossRefGoogle Scholar
  3. Brigl M, van den Elzen P, Chen X et al (2006) Conserved and heterogeneous lipid antigen specificities of CD1d-restricted NKT cell receptors. J Immunol 162:161–167.  https://doi.org/10.4049/jimmunol.176.6.3625 Google Scholar
  4. de Jong A (2015) Activation of human T cells by CD1 and self-lipids. Immunol Rev 267:16–29.  https://doi.org/10.1111/imr.12322 CrossRefGoogle Scholar
  5. Dearman RJ, Alcocer MJC, Kimber I (2007) Influence of plant lipids on immune responses in mice to the major Brazil nut allergen Ber e 1. Clin Exp Allergy 37:582–591.  https://doi.org/10.1111/j.1365-2222.2007.02689.x CrossRefGoogle Scholar
  6. Dowds CM, Kornell S-C, Blumberg RS, Zeissig S (2014) Lipid antigens in immunity. J Biol Chem 395:61–81.  https://doi.org/10.1515/hsz-2013-0220.Lipid Google Scholar
  7. Ghumra A, Alcocer M (2017) The use of a semi-automated system to measure mouse natural killer T (NKT) cell activation by lipid-loaded dendritic cells. Methods in molecular biology. Human Press, Clifton, pp 249–262Google Scholar
  8. Godfrey DI, Uldrich AP, Mccluskey J et al (2015) The burgeoning family of unconventional T cells. Nat Immunol 16:1114–1123.  https://doi.org/10.1038/ni.3298 CrossRefGoogle Scholar
  9. Guo XJ, Dash P, Calverley M et al (2016) Rapid cloning, expression, and functional characterization of paired αβ and γδ T-cell receptor chains from single-cell analysis. Mol Ther 3:15054.  https://doi.org/10.1038/mtm.2015.54 Google Scholar
  10. Heemskerk MHM, Hoogeboom M, De Paus RA et al (2003) Redirection of antileukemic reactivity of peripheral T lymphocytes using genetransfer of minor histocompatibility antigen HA-2-specific T-cell receptor complexes expressing a conserved alpha joining region. Blood 102:3530–3540.  https://doi.org/10.1182/blood-2003-05-1524 CrossRefGoogle Scholar
  11. Jordan-Williams KL, Poston S, Taparowsky EJ (2013) BATF regulates the development and function of IL-17 producing iNKT cells. BMC Immunol 14:16.  https://doi.org/10.1186/1471-2172-14-16 CrossRefGoogle Scholar
  12. Kean DE, Goodridge HS, McGuinness S et al (2006) Differential polarization of immune responses by plant 2S seed albumins, Ber e 1, and SFA8. J Immunol 177:1561–1566.  https://doi.org/10.4049/jimmunol.177.3.1561 CrossRefGoogle Scholar
  13. Kim PJ, Pai S-Y, Brigl M et al (2006) GATA-3 Regulates the development and function of invariant NKT cells. J Immunol 177:6650–6659.  https://doi.org/10.4049/jimmunol.177.10.6650 CrossRefGoogle Scholar
  14. Kim HS, Kim HS, Lee CW, Chung DH (2010) T cell Ig domain and mucin domain 1 engagement on invariant NKT cells in the presence of TCR stimulation enhances IL-4 production but inhibits IFN-γ production. J Immunol 184:4095–4106.  https://doi.org/10.4049/jimmunol.0901991 CrossRefGoogle Scholar
  15. Knies D, Klobuch S, Xue S-A et al (2016) An optimized single chain TCR scaffold relying on the assembly with the native CD3-complex prevents residual mispairing with endogenous TCRs in human T-cells. Oncotarget 7:21199–21221.  https://doi.org/10.18632/oncotarget.8385 CrossRefGoogle Scholar
  16. Lantz O, Bendelac A (1994) An invariant T cell receptor α 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 180:1097.  https://doi.org/10.1084/jem.180.3.1097 CrossRefGoogle Scholar
  17. Liu Z, Chen O, Wall JBJ et al (2017) Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Sci Rep 7:2193.  https://doi.org/10.1038/s41598-017-02460-2 CrossRefGoogle Scholar
  18. Masterson AJ, Sombroek CC, De Gruijl TD et al (2002) MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34 + precursors. Blood 100:701–703.  https://doi.org/10.1182/blood.V100.2.701 CrossRefGoogle Scholar
  19. Mirotti L, Florsheim E, Rundqvist L et al (2013) Lipids are required for the development of Brazil nut allergy: the role of mouse and human iNKT cells. Allergy Eur J Allergy Clin Immunol 68:74–83.  https://doi.org/10.1111/all.12057 CrossRefGoogle Scholar
  20. Ning J, Morgan D, Pamphilon D (2011) A rapid culture technique produces functional dendritic-like cells from human acute myeloid leukemia cell lines. J Biomed Biotechnol 2011:172965.  https://doi.org/10.1155/2011/172965 CrossRefGoogle Scholar
  21. O’Konek JJ, Terabe M, Berzofsky JA (2012) The role of NKT cells in the immune regulation of neoplastic disease. Innate immune regulation and cancer immunotherapy. Springer, New York, pp 7–21CrossRefGoogle Scholar
  22. Roth H, Magg V, Uch F et al (2017) Flavivirus infection uncouples translation suppression from cellular stress responses. MBio.  https://doi.org/10.1128/mBio.02150-16 Google Scholar
  23. Rundqvist L, Tengel T, Zdunek J et al (2012) Solution structure, copper binding and backbone dynamics of recombinant Ber e 1-the major allergen from Brazil Nut. PLoS ONE 7:46435.  https://doi.org/10.1371/journal.pone.0046435 CrossRefGoogle Scholar
  24. Sommermeyer D, Neudorfer J, Weinhold M et al (2006) Designer T cells by T cell receptor replacement. Eur J Immunol 36:3052–3059.  https://doi.org/10.1002/eji.200636539 CrossRefGoogle Scholar
  25. Taniguchi M, Seino KI, Nakayama T (2003) The NKT cell system: bridging innate and acquired immunity. Nat Immunol 4:1164–1165.  https://doi.org/10.1038/ni1203-1164 CrossRefGoogle Scholar
  26. Thomas S, Stauss HJ, Morris EC (2010) Molecular immunology lessons from therapeutic T-cell receptor gene transfer. Immunology 129:170–177.  https://doi.org/10.1111/j.1365-2567.2009.03227.x CrossRefGoogle Scholar
  27. Zhong S, Malecek K, Perez-Garcia A, Krogsgaard M (2010) Retroviral transduction of T-cell receptors in mouse T-cells. J Vis Exp 44:2307.  https://doi.org/10.3791/2307 Google Scholar
  28. Zhou Q, Buchholz CJ (2013) Cell type specific gene delivery by lentiviral vectors: new options in immunotherapy. Oncoimmunology 2:22566.  https://doi.org/10.4161/onci.22566 CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.School of BiosciencesUniversity of NottinghamLoughboroughUK
  2. 2.School of Life SciencesUniversity of NottinghamNottinghamUK
  3. 3.Unilever Safety and Environmental Assurance Centre (SEAC)SharnbrookUK

Personalised recommendations