Vγ4Vδ1 TCR Lentivirus

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78986

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Description

Vγ4Vδ1 TCR Lentivirus are replication incompetent, HIV-based, VSV-G-pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses transduce a TCR (T Cell Receptor) which belongs to the Vγ4Vδ1 subtype of γδ TCRsd. The TCR γ chain and δ chain are linked by P2A. The lentiviruses also transduce a puromycin selection marker (Figure 1).

Figure 1. (A) Schematic of the lenti-vector used to generate the Vγ4Vδ1 TCR Lentivirus and (B) diagram of the construct, showing the components of the Vγ4Vδ1 TCR.
TRGV and TRGC correspond to the TCR γ chain variable and constant regions, respectively, whereas TRDV and TRDC correspond to the TCR δ chain variable and constant regions.

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Product Data Gallery

Analysis of the expression of Vγ4Vδ1 TCR in TCR Knockout NFAT-Luciferase Reporter Jurkat Cells transduced with the Vγ4Vδ1 TCR Lentivirus

NFAT luciferase reporter activity in TCR Knockout NFAT-Luciferase Reporter Jurkat cells transduced with Vγ9Vδ2 TCR Lentivirus and Vγ4Vδ1 TCR Lentivirus and stimulated with agonist antibodies

Product Info

Storage and Usage

Citations

Supplied As

≥2 x 10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
Thaw Medium 2	BPS Bioscience #60184
Growth Medium 2B	BPS Bioscience #79530
Growth Medium 2F	BPS Bioscience #79669
TCR Knockout NFAT-Luciferase Reporter Jurkat Cell Line	BPS Bioscience #78556
Anti-CD3 Agonist Antibody	BPS Bioscience #71274
TCR Vδ1 Antibody, anti-human, PE, REAfinity™	Miltenyi Biotech #130-120-580
TCR V delta 2 Monoclonal Antibody (15D)	Life Technologies #TCR1732
TCR V delta 1 Monoclonal Antibody (TS8.2)	Life Technologies #TCR1730
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Lenti-Fuse Polybrene Viral Transduction Enhancer	BPS Bioscience #78939

Formulation

The lentiviruses were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

γδ TCRs (T cell receptors), αβ TCRs, and antibodies, result from gene rearrangements and offer the immune system the possibility to recognize several different types of antigens. γδ TCRs recognize antigens in a similar way to antibodies, being able to recognize full protein antigens and being independent on antigen binding to the MHC (major histocompatibility complex). γδ TCRs are cell type-specific, with Vγ4Vδ1 being present in γδTIL (tumor infiltrating lymphocytes) cells. Vδ1 expressing cells are found in mucosal and epithelial tissues and correspond to about 15% of the γδT cells present in PBMCs, with the % of γδ being only 5% of all the T cells. Vγ4Vδ1 cells can be activated by CD1 (cluster of differentiation 1) and respond to BTNL3 (butyrophilin-like 3), BTNL8, annexin A2 and A6. These cells can lead to tumor cell death by lysis. While most of the studies have been focusing on Vγ9Vδ2 T cells, a better understanding of the function and therapeutic potential of Vγ4Vδ1 T cells may open new avenues in cancer therapy.

References

Allison T. and Garboczi D., 2002 Molecular Immunology 38 (14): 1051-1061.
Song Y., et al., 2022 Front Immunol 13: 914839.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the virus at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Growth Media

For best results, the use of BPS Bioscience validated and optimized media is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for Cell Culture
Growth Medium 2B (BPS Bioscience #79530): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 1 mg/ml Geneticin.

Growth Medium 2F (BPS Bioscience #79669): RPMI1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 0.5 μg/ml of Puromycin and 1 mg/ml Geneticin.

Media Used in Functional Cellular Assay
Thaw Medium 2 (BPS Bioscience #60184): RPMI 1640 medium supplemented with 10% FBS, and 1% Penicillin/Streptomycin.

Applications

Use to transduce cells with the goal of screening Vγ4Vδ1 TCR agonist antibodies.
Use to transduce cells to be used as a positive control for Vγ4Vδ1 TCR evaluation and optimize experimental conditions.
Generate Vγ4Vδ1 TCR expressing cell pools or stable cell lines, following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a Vγ4Vδ1 TCR expressing stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection. 

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For further questions, please email [email protected].

Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions

Warnings

Avoid freeze/thaw cycles

Scientific Category

Immune Cell Signaling

Regulatory Status

Research Use Only

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Vγ4Vδ1 TCR Lentivirus

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