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Updated: Nov 7, 2025

Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers
Published on: March 6, 2009
Imaging and analysis on the interaction between human antigen-pulsed Vδ2 T cells and antigen-specific CD4 T cells
Yufei Mo1, Allen Ka Loon Cheung2, Yue Liu2
1AIDS 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.
Insights
This protocol visualizes surface protein interactions between γδ-T cells and CD4 T cells at their interface. It uses immunofluorescence and confocal microscopy to assess co-localization of proteins like PD1 and TLR4.
Area of Science:
- Immunology
- Cell Biology
- Microscopy
Background:
- Cell-cell interactions are crucial for immune responses.
- Understanding surface protein co-localization at the immune cell interface is vital.
- Current methods may lack resolution for detailed interface analysis.
Purpose of the Study:
- To establish a protocol for visualizing surface protein co-localization at the γδ-T cell and CD4 T cell interface.
- To enable the study of specific protein interactions, such as PD1 and TLR4, between these cell types.
Main Methods:
- Consolidation of immunofluorescence assay, confocal microscopy, and 3D imaging analysis.
- Co-culture of antigen-presenting γδ-T cells and CD4 T cells.
- Visualization of surface protein distribution at the cell-cell junction.
Main Results:
- Successful visualization of surface protein-protein co-localization at the cell-cell interface.
- Assessment of interaction between specific surface proteins (e.g., Δ42PD1, TLR4) in co-cultured cells.
- Demonstration of the protocol's applicability to various surface proteins and cell types.
Conclusions:
- The described protocol provides a robust method for studying protein interactions at immune cell interfaces.
- This technique facilitates a deeper understanding of cell-cell communication in immunological contexts.
- The protocol is adaptable for investigating novel protein interactions at cell junctions.
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).
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