A Structural Change in Butyrophilin upon Phosphoantigen Binding Underlies Phosphoantigen-Mediated Vγ9Vδ2 T Cell Activation

Yunyun Yang, Liping Li, Linjie Yuan, Xiaoying Zhou, Jianxin Duan, Hongying Xiao, Ningning Cai, Shuai Han, Xianqiang Ma, Weidong Liu, Chun Chi Chen, Lingle Wang, Xin Li, Jiahuan Chen, Ning Kang, Jing Chen, Zhixun Shen, Satish R. Malwal, Wanli Liu, Yan ShiEric Oldfield, Rey Ting Guo, Yonghui Zhang

Research output: Contribution to journalArticle


Human Vγ9Vδ2 T cells respond to microbial infections and malignancy by sensing diphosphate-containing metabolites called phosphoantigens, which bind to the intracellular domain of butyrophilin 3A1, triggering extracellular interactions with the Vγ9Vδ2 T cell receptor (TCR). Here, we examined the molecular basis of this “inside-out” triggering mechanism. Crystal structures of intracellular butyrophilin 3A proteins alone or in complex with the potent microbial phosphoantigen HMBPP or a synthetic analog revealed key features of phosphoantigens and butyrophilins required for γδ T cell activation. Analyses with chemical probes and molecular dynamic simulations demonstrated that dimerized intracellular proteins cooperate in sensing HMBPP to enhance the efficiency of γδ T cell activation. HMBPP binding to butyrophilin doubled the binding force between a γδ T cell and a target cell during “outside” signaling, as measured by single-cell force microscopy. Our findings provide insight into the “inside-out” triggering of Vγ9Vδ2 T cell activation by phosphoantigen-bound butyrophilin, facilitating immunotherapeutic drug design.

Original languageEnglish (US)
Pages (from-to)1043-1053.e5
Issue number4
StatePublished - Apr 16 2019



  • T cell antigen recognition
  • Vγ9Vδ2 T cells
  • allogenic cell therapy
  • butyrophilin 3A1
  • cell therapy
  • immune stimulant
  • immunotherapy
  • inside-out signaling
  • phosphoantigens

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Infectious Diseases

Cite this

Yang, Y., Li, L., Yuan, L., Zhou, X., Duan, J., Xiao, H., Cai, N., Han, S., Ma, X., Liu, W., Chen, C. C., Wang, L., Li, X., Chen, J., Kang, N., Chen, J., Shen, Z., Malwal, S. R., Liu, W., ... Zhang, Y. (2019). A Structural Change in Butyrophilin upon Phosphoantigen Binding Underlies Phosphoantigen-Mediated Vγ9Vδ2 T Cell Activation. Immunity, 50(4), 1043-1053.e5. https://doi.org/10.1016/j.immuni.2019.02.016