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Adenosine A2A Receptor Functional Recombinant Cell Line

Adenosine A2A Receptor Functional Recombinant Cell Line

Catalog #: 79381

Size: 2 vials

$8,500

Background:

Adenosine signaling plays an important role in inflammation and the immune response. Many cells in the tumor microenvironment express ectopic CD39 and CD73, leading to the buildup of extracellular adenosine. Engagement of adenosine with the high affinity A2a receptor (A2aR) on the surface of T cells, macrophages, NK cells, neutrophils, and dendritic cells causes downregulation of the immune response. Therefore, A2aR is a novel immune checkpoint protein, and blockade of A2aR is being actively investigated as a potential immunotherapy.

Several A2aR antagonists have progressed to clinical trials for the treatment of Parkinson’s disease, and preclinical studies have confirmed that blockade of A2a receptor activation has the ability to markedly enhance anti-tumor immunity. Mice treated with A2aR antagonists, such as ZM241385 (see data below) or caffeine, show significantly delayed tumor growth, and A2aR knockout mice demonstrate increased tumor rejection. Most promising, A2aR blockade can be used in synergy with the inhibition of other immune checkpoint pathways. Studies show that the combination of A2aR blockade and PD-1 inhibition is more effective than either treatment separately, and A2aR blockade increases the activity of CTLA-4 and TIM-3 inhibition in controlling the growth of CD73+ melanoma.

Description:

Adenosine A2a receptor (A2aR or ADORA2A) stably expressed in HEK-293 reporter cells with a cAMP-responsive luciferase construct (CRE/CREB Luc, BPS Bioscience, #60515). A2aR is a member of the seven transmembrane G protein-coupled receptor (GPCR) family. The activity of A2aR is mediated by Gs protein which activates adenylyl cyclase, resulting in the synthesis of intracellular cAMP.

The level of cAMP correlates with the respective adenosine (agonist) level, and the cell line can be used to measure the EC50 and IC50 values of A2aR agonists or antagonists in a quantitative manner. Binding of extracellular adenosine ligand or its stable analog NECA (5′-(N-Ethylcarboxamido) adenosine) to the A2aR expressed on the surface of this cell line can also produce luciferase reporter signal for qualitative measurement.

Materials Required But Not Provided:

• Assay Medium: Thaw Medium 1 (BPS Bioscience, #60187)

• NECA (Sigma, #79262)

• ZM241385 (Sigma, #76263)

• 96-well tissue culture-treated white clear-bottom assay plate

• cAMP Glo kit (Promega, #V1501)

• One-Step luciferase assay system (BPS Bioscience, #60690) for measuring firefly luciferase activity

• Luminometer

Synonym(s):

A2aR, ADORA2A

Supplied As:

Each vial contains ~ 2 x 106 cells in 1 ml of 10% DMSO in FBS.

Host Cell line:

HEK293 cells

Mycoplasma Testing:

This cell line has been screened using the MycoAlert™ Mycoplasma Detection Kit (Lonza, #LT07-118) to confirm the absence of Mycoplasma contamination. MycoAlert Assay Control Set (Lonza, #LT07-518) was used as a positive control.

Growth Media:

Thaw Medium 1 (BPS Bioscience, #60187): MEM medium (Hyclone, #SH30024.01) + 10% FBS (Life Technologies, #26140-079) + 1% non-essential amino acids (Hyclone, #SH30238.01) + 1 mM Na pyruvate (Hyclone, #SH30239.01) + 1% Penicillin/Streptomycin (Hyclone, SV30010.01)

Complete Growth Medium: Thaw Medium 1 (BPS Bioscience, #60187) plus 0.4 mg/ml G418 (Thermo Fisher, # 11811031) and 50 μg/ml of Hygromycin B (Hyclone, #SV30070.01)

Cells should be grown at 37°C with 5% CO2 using complete growth medium (Thaw Medium 1 with G418 and Hygromycin B).

Instructions for use:

See cell line data sheet for detailed culturing and assay protocol.

Storage / Stability:

Immediately upon receipt, store in liquid nitrogen.

Application(s):

• Screen for agonists or antagonists of A2aR in cell-based cAMP assays.

• Study PD-1 and CTLA-4 combination therapy.

• Screen co-inhibitor immune checkpoint molecules for cancer immunotherapy.

Reference(s):

1. Leone, R.D., et.al. (2015). Comp. Struct. Biotechnol. J. 13: 265-272

2. Ma, S-R., et al. (2017). Molec. Cancer 16:99

3. Ohta, A., et al. (2016). Front. Immunol. 7:109.

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 the 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.

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Quality Assurance

Product Datasheets

79381.pdf