Related Experiment Video
Updated: Aug 15, 2026
Isolation and Ex Vivo Culture of Vδ1+CD4+γδ T Cells, an Extrathymic αβT-cell Progenitor
Published on: December 7, 2015
T cell receptor-alpha beta lacking the beta-chain V domain can be expressed at the cell surface but prohibits T cell
F Ossendorp1, H Jacobs, G van der Horst
1Division of Immunology, The Netherlands Cancer Institute, Amsterdam.
Abstract:
A TCR-beta gene lacking V domain sequences (delta V-TCR-beta) was inserted into the germline of mice. Expression of the transgene inhibited endogenous TCR-beta, but not TCR-alpha gene rearrangement and expression. The mutated TCR-beta gene affected alpha beta T cell development: the common thymocyte pool was normal in cell number, with cells expressing CD4 and CD8, but the mature, "CD3bright" population expressing either CD4 or CD8 molecules was reduced by 90%. To help understand these effects on TCR-beta gene rearrangement and T cell development, biosynthesis of the delta V-TCR-beta protein was analyzed in a tumor cell line derived from a transgenic mouse. Despite absence of the V domain, the delta V-TCR-beta chain paired with endogenous TCR-alpha chains and assembled with CD3 gamma, -delta, -epsilon, and -zeta components in the endoplasmatic reticulum, followed by transport through the Golgi complex to the plasma membrane. Therefore, assembly of the complex, and even cell surface expression, may be relevant for allelic exclusion of the TCR-beta gene. In the common thymocyte population, the CD3 components, endogenous TCR-alpha, and the delta V-TCR-beta gene product were expressed at the RNA level, but endogenous TCR-beta was not. The TCR-alpha delta beta/CD3 complex was present at the cell surface at low levels and was functional in terms of anti-CD3-induced Ca2+ mobilization. The observed arrest of alpha beta T cell development at the CD4+8+ thymocyte stage indicates that ligand recognition by the TCR, with contribution of the beta-chain V domain, is not required for transition of CD4-8- thymocytes to the CD4+8+ phenotype, but necessary for entry into the "single positive," CD3bright differentiation stage.
Insights
Transgenic mice with a mutated T-cell receptor beta (TCR-beta) gene showed impaired T-cell development. This suggests the TCR-beta V domain is crucial for mature T-cell differentiation and proper immune responses.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- T-cell receptor (TCR) gene rearrangement is essential for T-cell development and immune function.
- The V domain of the TCR-beta chain plays a critical role in T-cell recognition and signaling.
Purpose of the Study:
- To investigate the impact of a truncated TCR-beta gene (delta V-TCR-beta) on T-cell development and TCR gene rearrangement in mice.
- To elucidate the role of the TCR-beta V domain in T-cell maturation and allelic exclusion.
Main Methods:
- Generation of transgenic mice expressing a delta V-TCR-beta gene.
- Analysis of T-cell development using flow cytometry (CD4, CD8, CD3 expression).
- Investigation of protein biosynthesis, assembly, and cell surface expression of the delta V-TCR-beta chain.
Main Results:
- Expression of delta V-TCR-beta inhibited endogenous TCR-beta rearrangement and expression but not TCR-alpha.
- Transgenic mice exhibited a 90% reduction in mature CD3bright T cells (CD4+ or CD8+).
- The delta V-TCR-beta chain assembled with TCR-alpha and CD3 components, reaching the cell surface and mediating signaling.
Conclusions:
- The TCR-beta V domain is not required for initial T-cell development to the CD4+8+ stage.
- The V domain is essential for the transition to the mature CD3bright single-positive stage.
- Cell surface expression of the assembled TCR complex, even without the V domain, may contribute to TCR-beta allelic exclusion.
More Related Videos
08:48Generating De Novo Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain
Published on: October 25, 2016
11:21Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
Published on: September 10, 2017
Related Concept Videos
02:38Mitogens and the Cell Cycle
01:15Receptor Downregulation in MVBs
The EGFR can initiate signaling pathways that lead to cell proliferation, migration, and differentiation. Overexpression of EGFR stimulates cells to proliferate. Excessive EGFR activation may...
01:28Diversity of Antigen Receptors
Before encountering any antigen, lymphocytes express these receptors. On B cells, the antigen receptor is a membrane-bound antibody molecule called BCR; on T cells, it is a T cell receptor or TCR. B and T cell receptors are composed of two...
01:22T Cell Activation and Clonal Selection
Naive T cells that have not yet encountered an antigen express two primary CD...
01:27Cytotoxic T Cells-mediated Immune Response
Immunological surveillance is the ability of immune cells to monitor and eliminate infected cells with intracellular pathogens, neoplastically transformed cells, and cells with non-self antigens. Cytotoxic T cells and NK...
01:24B Cell Activation and Differentiation
When naive B cells encounter a specific antigen that can bind to the B cell receptor (BCR) on their surface, they undergo sensitization to respond to the antigen's presence. Sensitization begins with...