Imaging and analysis on the interaction between human antigen-pulsed Vδ2 T cells and antigen-specific CD4 T cells

Yufei Mo; Allen Ka Loon Cheung; Yue Liu; Li Liu; Zhiwei Chen

doi:10.1016/j.xpro.2021.100453

Imaging and analysis on the interaction between human antigen-pulsed Vδ2 T cells and antigen-specific CD4 T cells

STAR Protoc. 2021 Apr 14;2(2):100453. doi: 10.1016/j.xpro.2021.100453. eCollection 2021 Jun 18.

Authors

Yufei Mo¹, Allen Ka Loon Cheung², Yue Liu², Li Liu¹, Zhiwei Chen¹

Affiliations

¹ AIDS Institute and Department of Microbiology, State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China.
² Department of Biology, Faculty of Science, Hong Kong Baptist University, Hong Kong SAR, China.

Abstract

This protocol describes how to visualize surface protein-protein co-localization across a cell-cell interface between antigen-presenting γδ-T cells and CD4 T cells. By consolidating immunofluorescence assay, confocal microscopy and 3D imaging analysis, it enables assessment of interaction between cell surface proteins such as Δ42PD1 and TLR4 between co-cultured γδ-T and CD4 T cells. This protocol can be applied to study a surface protein of interest and its potential interaction with a target cell/protein at the cell-cell interface. For complete details on the use and execution of this profile, please refer to Mo et al. (2020).

Keywords: Cell culture; Cell isolation; Immunology; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CD4-Positive T-Lymphocytes* / cytology
CD4-Positive T-Lymphocytes* / immunology
Coculture Techniques
Humans
Imaging, Three-Dimensional*
Microscopy, Confocal
Receptors, Antigen, T-Cell, gamma-delta / immunology*
Toll-Like Receptor 4 / immunology*

Substances

Receptors, Antigen, T-Cell, gamma-delta
TLR4 protein, human
Toll-Like Receptor 4