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Expression, Purification, and Crystallization of the Vγ9Vδ2 T-cell Receptor Recognizing Protein/Peptide Antigens

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Abstract

γδ T cells, especially Vγ9Vδ2 T cells, play an important role in mycobacterial infection. We have identified some Vγ9Vδ2 T cells that recognize protein/peptide antigens derived from mycobacteria, which may induce protective immune responses to mycobacterial infection. To clarify the structural basis of the molecular recognition mechanism, we tried many methods to express the Vγ9Vδ2 T-cell receptor (TCR). The Vγ9Vδ2 TCR was not expressed well in a prokaryotic expression system or a baculovirus expression system, even after extensive optimization. In a mammalian cell expression system, the Vγ9Vδ2 TCR was expressed in the form of a soluble heterodimer, which was suitable for crystal screening. Reduced-temperature cultivation (cold shock) increased the yield of the recombinant TCR. The recombinant purified TCR was used for crystal trials, and crystals that could be used for X-ray diffraction were obtained. Although we have not yet determined the crystal structure of the Vγ9Vδ2 TCR, we have established a procedure for Vγ9Vδ2 TCR expression and purification, which is useful for basic research and potentially for clinical application.

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Acknowledgements

We wish to dedicate this work to the memory of Pro. Zhendong Zhao. We are grateful to Pro. Jennifer Maynard (University of Texas at Austin) and Prof. Jamie Rossjohn (Monash University) for helpful discussion. This work was supported by the Key Science and Technology Program of Henan Province (Grant No. 222102310626 and 232102310346). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Stem Cell Research Center, Henan Key Laboratory of Stem Cell Differentiation and Modification, Henan Provincial People’s Hospital, People’s Hospital of Zhengzhou University, Zhengzhou, 450003, China

    Chaofei Cheng & Guangzhi Liu

  2. People’s Hospital of Henan University, Zhengzhou, 450003, China

    Chaofei Cheng & Guangzhi Liu

  3. MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Centre for Tuberculosis Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100730, China

    Chaofei Cheng & Zhendong Zhao

  4. Clinical Immunology Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100730, China

    Zhendong Zhao

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  1. Chaofei Cheng
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Z.Z. and G.L. conceptualized and designed the project. C.C. performed the experiments. C.C. wrote the manuscript. All authors contributed to the discussion and reviewed the manuscript.

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Correspondence to Zhendong Zhao or Guangzhi Liu.

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Cheng, C., Zhao, Z. & Liu, G. Expression, Purification, and Crystallization of the Vγ9Vδ2 T-cell Receptor Recognizing Protein/Peptide Antigens. Protein J 42, 778–791 (2023). https://doi.org/10.1007/s10930-023-10151-3

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