αβ TCR Generation for Engineered γδ T Cell

γδ T cells span the innate and adaptive arms of the immune system and participate in the response to pathogens and tumors. Similar to αβ T cells, γδ T cells develop in thymus, but express rearranged T cell receptor (TCR) composed of TCR-γ and TCR-δ chains.

Background

The successful application of T cell-based immunotherapy depends on the availability of a large number of T cells with the desired antibody (Ag) specificity and phenotypic characteristics. The engineering of TCR transferred T lymphocytes is an attractive strategy to obtain enough Ag-specific T cells. However, due to the formation of mixed dimers between the endogenous and the introduced TCR chain, the introduction of additional TCR chains into T cells will lead to the production of T cells with unknown specificity. Previous studies have shown that the γ and δ TCR chains expressed by γδ T cells cannot form heterodimers with α and β TCR chains. Therefore, using γδ T cells as receptor cells can prevent the formation of such potentially self-aggressive T cells. Some studies have shown that αβ TCR engineering of γδ T cells can form a feasible strategy for Ag specific effector T cells that do not express mixed TCR dimers.

Fig.1 Genetic modification of γδ T cells for adoptive therapy approaches to cancer. (Legut, 2015)

αβ TCR Generation for Engineered γδ T Cell Service

At present, it is proved in cancer research that cancer-specific αβ or γδ TCR cannot compete well with receptor TCR, so it shows weak functional activity. If it is combined with endogenous TCR-β and knocked out at the same time, receptor T cells can be effectively redirected to cancer. Therefore, the mismatch between the endogenous TCR-α chain and transduced TCR-β of the constructed engineered T cell must be minimal. In cancer, our design may be like this: in combination with endogenous TCR gene knockout, using zinc finger nuclease, transcription activator-like effector nuclease or CRISPR/Cas9 to redirect T cells to cancer through chimeric antigen receptors (CARs) in most cases.

Fig.2 Construct design and validation for transduction of primary T-cells. (Legut, 2018)

This method can also perform a detailed analysis of TCR recognition in the absence of parental T cell clones. This TCR may come from T cells with poor growth characteristics (e.g., due to cancer-mediated T cell failure), or directly from high-throughput sequencing of TCR sequences. We can also imagine that primary T cells have unknown specificity, but do not express endogenous TCR, and can be used for high-throughput genome-wide screening to identify new TCR ligands, thereby identifying new potential therapeutic targets.

Features and Advantages

Based on the general method of TCR construction, Creative Biolabs provides an optimized αβ TCR generation for engineered γδ T cell vector production method.

• Rich experience in the preparation of plasmid DNA.
• High purity and endotoxin free plasmids were provided.
• Be able to customize the design and structure of γδ T cell products for customers according to their special needs.

Scientists at Creative Biolabs are committed to being a reliable partner for your γδ T cell design, construction and plasmid production, and providing the most effective solutions for your projects. If you want to get more information, please feel free to contact us.

References

1. Legut, M., et al. The promise of γδ T cells and the γδ T cell receptor for cancer immunotherapy. Cell Mol Immunol. 2015, 12: 656-668.
2. Legut, M., et al. CRISPR-mediated TCR replacement generates superior anticancer transgenic T cells. Blood. 2018, 131(3): 311-322.