BTLA / NFAT - Luciferase Reporter - Jurkat Recombinant Cell Line

Catalog #: 79476
Size: 2 Vials
$8,000
Background:

BTLA is an immunoglobulin domain-containing glycoprotein expressed on T cells, resting B cells, macrophages, DCs and, to a lesser extent, NK cells. BTLA acts as an inhibitory receptor on T cells, as anti-BTLA treatment results in T cell proliferation, and BTLA knockout mice demonstrate hyper-responsive immune activation. Subsequently, herpesvirus entry mediator (HVEM), a tumor necrosis factor receptor, was identified as a natural ligand for BTLA in mice and humans. Expression of HVEM by antigen-presenting cells (APCs) was capable of inducing BTLA-dependent T cell inhibition.

BTLA belongs to the immunoglobulin superfamily, along with CTLA-4 and PD-1, which characteristically binds to B7 family members. HVEM is a member of the tumor necrosis factor receptor family, and the BTLA/HVEM interaction is the first to demonstrate crosstalk between these two receptor families. Prior to the discovery of the BTLA/HVEM interaction, HVEM was known to bind LIGHT and lymphotoxin-α, both tumor necrosis factor ligands. While the BTLA/HVEM interaction results in a co-inhibitory signal, the HVEM/LIGHT interaction results in a co-stimulatory signal through HVEM.

In melanoma patients, BTLA was demonstrated to be expressed on tumor-specific T cells both in circulation and in metastatic lymph nodes. HVEM was subsequently shown to be expressed in the majority of cultured melanoma cell lines and metastatic melanoma samples. The interaction of BTLA on tumor specific T cells and HVEM on melanoma cells resulted in T cell inhibition, which could be reversed by treatment with a BTLA blocking antibody. BTLA/HVEM interaction plays a key role in the regulation of inflammatory, autoimmune, and antitumor responses, and is an important target for cancer immunotherapy drug discovery. Therapeutically targeting BTLA and HVEM remains in pre-clinical stages as the bidirectional signaling pathways of BTLA/HVEM and HVEM/LIGHT are further elucidated.

Description:

Recombinant Jurkat T cell expressing firefly luciferase gene under the control of NFAT response elements with constitutive expression of human BTLA (B and T lymphocyte associated, B and T lymphocyte attenuator, BTLA1, CD272, GenBank Accession #NM_181780).

Materials Required But Not Provided:
  • • CHO-K1 cell and its growth medium
  • • TCR Activator/HVEM Mammalian Expression Kit (BPS Bioscience, coming soon), including TCRa/pIRES expression vector and HVEM/pIRES expression vector
  • • HVEM/CHO_TCR activator stable cell line (BPS Bioscience, coming soon)
  • • Transfection reagent for generating target cells (aka artificial Antigen Presenting Cells (aAPC)) [We use Lipofectamine™ 2000 (Life technologies #11668027). However, other transfection reagents work equally well.]
  • • Opti-MEM I Reduced Serum Medium (Life technologies #31985-062)
  • • Assay Medium: Thaw Medium 2 (BPS Cat. #60184)
  • • Anti-BTLA neutralizing antibody (for testing)
  • • Anti-HVEM neutralizing antibody (for testing
  • • 96-well tissue culture treated white clear-bottom assay plate (Corning, #3610)
  • ONE-Step™ Luciferase Assay System (BPS Bioscience, #60690)
  • • Luminometer
Synonym(s):
B and T lymphocyte associated, B and T lymphocyte attenuator, BTLA1, CD272
Supplied As:
Each vial contains 2 x 10^6 cells in 1 ml of 10% DMSO.
Host Cell line:
Jurkat
Mycoplasma Testing:

The cell line has been screened using the PCR-based Venor®GeM Mycoplasma Detection kit (Sigma-Aldrich, #MP0025) to confirm the absence of Mycoplasma species.

Growth Media:

Thaw Medium 2 (BPS Cat. #60184): RPMI1640 medium (Life Technologies #A10491-01) supplemented with 10% FBS (Life Technologies #26140-079), 1% Penicillin/Streptomycin (Hyclone #SV30010.01).

Complete Growth Medium: Thaw Medium 2 plus 1 mg/ml of Geneticin (Life Technologies #11811031) and 200 μg/ml of Hygromycin B (Hyclone #SV30070.01).

Cells should be grown at 37°C with 5% CO2 using complete growth medium (Thaw Medium 2 plus Geneticin and Hygromycin B).

Instructions for use:
See data sheet for detailed culturing and assay protocol.
Storage / Stability:
Immediately upon receipt, store vials in liquid nitrogen.
Application(s):
• Screen for activators or inhibitors of BTLA/HVEM interaction in cell-based co-inhibitory bioassay

• Characterize the biological activity of BTLA and its interactions with ligands

Reference(s):

1. BTLA marks a less cytotoxic T-cell subset in diffuse large B-cell lymphoma with high expression of checkpoints. Quan L. et al. Exp Hematol. 2018 Feb 3.

3. Design of short peptides to block BTLA/HVEM interactions for promoting anticancer Tcell responses. Spodzieja M, et al. PLoS One. 2017 Jun 8; 12(6): e0179201

Notes:

Contact us for pricing.

License Disclosure: Purchase of this cell line grants you with a 10-year license to use this cell line in your immediate laboratory, for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit use of this cell line in humans or for therapeutic or drug use. The license does not permit modification of the cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of BPS products to your laboratory. BPS does not warrant the suitability of the cell line for any particular use, and does not accept any liability in connection with the handling or use of the cell line. Modifications of this cell line, transfer to another facility, or commercial use of the cells may require a separate license and additional fees; contact sales@bpsbioscience.com for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.

Warning(s):
Avoid freeze/thaw cycles.
Scientific Category:
Immunotherapy
Product Type:
Cell Line
Data shown is lot-specific. Contact us for specific information on other lots.

Quality Assurance

Product Datasheets

79476.pdf