TIGIT / NFAT Reporter - Jurkat Cell Line

Catalog #: 60538
Size: 2 vials
$8,000
Background:
TIGIT is a co-inhibitory receptor that is highly expressed in Natural Killer (NK) cells, activated CD4+, CD8+ and regulatory T cells. Interaction with the poliovirus receptor (PVR; CD155) on antigen presenting cells, such as dendritic cells, recruits Src homology (SH) domain-containing protein tyrosine phosphatase SHP1 and SHP2 or the inositol phosphatase SHIP1 and SHIP2 to the TIGIT ITIM domain. This increases IL-10 release and suppresses NF-kB and NFAT T cell receptor (TCR) signaling, which blocks T cell proliferation and cytokine production. It serves as a competitive inhibitor of CD226, a co-stimulatory receptor for CD155. TIGIT targeting antibodies that block this T cell-intrinsic inhibitory effects have shown enhanced anti-tumor and anti-viral functions in preclinical studies.
Description:
Recombinant Jurkat T cell expressing firefly luciferase gene under the control of NFAT response elements with constitutive expression of human TIGIT (V-set and immunoglobulin domain-containing protein 9, VSIG9, V-set and transmembrane domain-containing protein 3, and VSTM3, GenBank Accession No. NM_173799).

TIGIT / NFAT Reporter - Jurkat Cell Line

Materials Required But Not Provided:

For Culture:

Thaw Medium 2 (BPS Cat. #60184)
Growth Medium 2A (BPS Cat. #60190)

For Assay:

• CD155/TCR Activator-CHO cell line (BPS Cat. # 60548)
• Assay Medium 2C (BPS Cat. #79623)
• Thaw Medium 3 (BPS Cat. #60186): Ham’s F-12 medium (Hyclone, #SH30526.01) supplemented with 10% FBS (Life Technologies #26140-079), 1% Penicillin/Streptomycin (Hyclone, #SV30010.01)
• Anti-TIGIT neutralizing antibody (BPS Cat. # 71340)
• 96-well tissue culture-treated white clear-bottom assay plate
• One-Step luciferase assay system (BPS Cat. # 60690) or other luciferase reagents for measuring firefly luciferase activity
• Luminometer

Synonym(s):
T-cell immunoreceptor with Ig and ITIM domains, V-set and immunoglobulin domain-containing protein 9, VSIG9, V-set and transmembrane domain-containing protein 3, VSTM3
Host Cell line:
Jurkat
Mycoplasma Testing:
The cell line has been screened using the PCR-based Venor®GeM Mycoplasma Detection kit (Sigma-Aldrich) 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). 

Growth Medium 2A (BPS Cat. #60190): Thaw Medium 2 (BPS Cat. #60184) + 1 mg/ml of Geneticin (Life Technologies #11811031) and 200 μg/ml of Hygromycin B (Hyclone #SV30070.01). 

Instructions for use:
See product datasheet for detailed protocol.
Storage / Stability:
Store in liquid nitrogen immediately upon receipt.
Application(s):
Screen for activators or inhibitors of TIGIT signaling in a cellular context, characterize the biological activity of TIGIT and its interactions with ligands
Reference(s):

1. Li M et.al. (2014) T-cell Immunoglobulin and ITIM Domain (TIGIT) Receptor/ Poliovirus Receptor (PVR) Ligand Engagement Suppresses Interferon-g Production of Natural Killer Cells via b-arrestin 2-mediated Negative Signaling. J Biol Chem. 289: 17647- 17657.
2. Stanietsky N. et.al. (2009) The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity. PNAS. 106: 17858-17863.
3. Zhu et.al. (2016) Identification of CD112R as a novel checkpoint for human T cells. J Exp. Med. 213: 167-176.
4. Yu X. et.al. (2008) The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat.Immunol. 10: 48-57.

Notes:

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

60538.pdf