Imaging and analysis on the interaction between human antigen-pulsed Vδ2 T cells and antigen-specific CD4 T cells
| Autor: | Yue Liu, Yufei Mo, Li Liu, Zhiwei Chen, Allen Ka Loon Cheung |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
CD4-Positive T-Lymphocytes
Science (General) Cell Immunology Immunofluorescence General Biochemistry Genetics and Molecular Biology law.invention Q1-390 Imaging Three-Dimensional Antigen Antigen specific Confocal microscopy law Microscopy medicine Protocol Humans Microscopy Confocal General Immunology and Microbiology medicine.diagnostic_test Chemistry General Neuroscience Receptors Antigen T-Cell gamma-delta Coculture Techniques Cell biology Toll-Like Receptor 4 medicine.anatomical_structure Cell culture Cell isolation TLR4 |
| Zdroj: | STAR Protocols STAR Protocols, Vol 2, Iss 2, Pp 100453-(2021) |
| ISSN: | 2666-1667 |
| Popis: | Summary 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). Graphical abstract Highlights • Protocol to co-culture antigen-pulsed Vδ2 T cell and antigen-specific CD4 T cell • Use of IFA for visualizing colocalization of proteins across cell-cell interface • Detailed procedures to perform Z-stack and 3D imaging analysis 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. |
| Databáze: | OpenAIRE |
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