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
Journal	STAR Protocols
Volume	2
Issue number	2
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.). ",

year = "2021",

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

language = "English",

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journal = "STAR Protocols",

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AU - Mo, Yufei

AU - CHEUNG, Ka Loon Allen

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

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