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

Yufei Mo; Ka Loon Allen 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

Yufei Mo, Ka Loon Allen CHEUNG^*, Yue Liu, Li Liu, Zhiwei Chen

^*Corresponding author for this work

Department of Biology

Research output: Contribution to journal › Article › peer-review

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).

Original language	English
Article number	100453
Number of pages	20
Journal	STAR Protocols
Volume	2
Issue number	2
Early online date	14 Apr 2021
DOIs	https://doi.org/10.1016/j.xpro.2021.100453
Publication status	Published - 18 Jun 2021

Scopus Subject Areas

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)

User-Defined Keywords

Cell culture
Cell isolation
Immunology
Microscopy

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10.1016/j.xpro.2021.100453

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

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",

author = "Yufei Mo and CHEUNG, {Ka Loon Allen} and Yue Liu and Li Liu and Zhiwei Chen",

note = "Funding Information: We thank Jingjing Li and Dongyan Zhou for generating the anti-Δ42PD1 monoclonal antibodies that were used in this study. This work was supported by Hong Kong Research Grant Council TRS-T11-706/18-N , GRF17115818 , GRF17104919 , GRF17103514 , and HKU5/CRF/13G , University Development Fund of the University of Hong Kong to AIDS Institute and the National Science and Technology Major Project 2018ZX10731-101-002-001 (to Z.C.), and Health and Medical Research Fund ( HMRF ) ( 15140732 and 18170032 ), Interdisciplinary Research Matching Scheme ( RC-IRCs-1718-03 ), Faculty Research Grant ( FRG2/17-18/066 ), Faculty Start-up Fund ( SCI-17-18-01 ), Tier 2 Start-up Grant ( RC-SGT2/18-19/SCI/007 ) (to A.K.L.C.). ",

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doi = "10.1016/j.xpro.2021.100453",

language = "English",

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AU - Liu, Yue

AU - Liu, Li

AU - Chen, Zhiwei

N1 - Funding Information: We thank Jingjing Li and Dongyan Zhou for generating the anti-Δ42PD1 monoclonal antibodies that were used in this study. This work was supported by Hong Kong Research Grant Council TRS-T11-706/18-N , GRF17115818 , GRF17104919 , GRF17103514 , and HKU5/CRF/13G , University Development Fund of the University of Hong Kong to AIDS Institute and the National Science and Technology Major Project 2018ZX10731-101-002-001 (to Z.C.), and Health and Medical Research Fund ( HMRF ) ( 15140732 and 18170032 ), Interdisciplinary Research Matching Scheme ( RC-IRCs-1718-03 ), Faculty Research Grant ( FRG2/17-18/066 ), Faculty Start-up Fund ( SCI-17-18-01 ), Tier 2 Start-up Grant ( RC-SGT2/18-19/SCI/007 ) (to A.K.L.C.).

PY - 2021/6/18

Y1 - 2021/6/18

N2 - 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).

AB - 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).

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

Abstract

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