CD27/TNFRSF7 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

CD27, also referred to as tumor necrosis factor receptor superfamily, member 7 (TNFRSF7), is a type I transmembrane glycoprotein that functions as a co-stimulatory T cell receptor and is expressed on the surface of T cells, natural killer (NK) cells, and B cells (1,2). The human CD27 protein is 260 amino acids (aa) in length and consists of a 19 aa signal sequence, 172 aa extracellular domain (ECD) containing three characteristic cysteine-rich domains (CRDs), a 21 aa helical transmembrane region, and a 48 aa cytoplasmic tail domain (3,4). The CD27 protein has a theoretical molecular weight (MW) of 29 kDa, but is typically is closer to 50-55 kDa due to N-linked and O-linked glycosylation (3). Mouse CD27 cDNA encodes a 250 aa protein with a theoretical molecular weight of 28 kDa (5). Human CD27 shares ~64% aa sequence identity with mouse CD27 protein.

Membrane-bound CD27 is expressed as a disulfide-linked homodimer (3). CD27 binds to the ligand CD70, a transmembrane glycoprotein that is transiently expressed on activated immune cells such as antigen presenting cells (APCs), dendritic cells (DCs), NK cells, B cells, and T cells (1,2,6,7). The receptor-ligand binding interaction leads to NFkappaB and c-Jun pathway activation which promotes immune stimulation and activation and survival of CD4+ T cells, CD8+ T cells, memory T cells, and NK cells (2,6,7). Both CD27 and CD70 are often abnormally expressed or dysregulated on malignant and cancer cells leading to immune evasion and tumor progression (7). CD27 has become a target of interest of immunotherapies for viral infections, autoimmune disease, and cancer (2). Varlilumab, an agonistic CD27 monoclonal antibody (mAB), has entered clinical trials for the treatment of hematological and solid tumor cancers (1,6). Additional clinical trials are in process that combine varlilumab with other immune checkpoint inhibitors like the programmed cell death protein-1 (PD-1) blocking mAb nivolumab (1,2). Initial results are promising, suggesting that targeting CD27, especially in combination with other therapeutics, may be a promising and effective immunotherapy for a variety of pathologies (1,2,6).

References

1. Starzer AM, Berghoff AS. New emerging targets in cancer immunotherapy: CD27 (TNFRSF7). ESMO Open. 2020;4(Suppl 3):e000629. https://doi.org/10.1136/esmoopen-2019-000629

2. Grant EJ, Nussing S, Sant S, Clemens EB, Kedzierska K. The role of CD27 in anti-viral T-cell immunity. Curr Opin Virol. 2017;22:77-88. https://doi.org/10.1016/j.coviro.2016.12.001

3. Buchan SL, Rogel A, Al-Shamkhani A. The immunobiology of CD27 and OX40 and their potential as targets for cancer immunotherapy. Blood. 2018;131(1):39-48. https://10.1182/blood-2017-07-741025

4. Uniprot (P26842)

5. Uniprot (P41272)

6. van de Ven K, Borst J. Targeting the T-cell co-stimulatory CD27/CD70 pathway in cancer immunotherapy: rationale and potential. Immunotherapy. 2015;7(6):655-667. https://doi.org/10.2217/imt.15.32

7. Flieswasser T, Van den Eynde A, Van Audenaerde J, et al. The CD70-CD27 axis in oncology: the new kids on the block. J Exp Clin Cancer Res. 2022;41(1):12. https://doi.org/10.1186/s13046-021-02215-y

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-08927

• CD27/TNFRSF7 Antibodies
• CD27/TNFRSF7 Lysate
• CD27/TNFRSF7 Protein
• CD27/TNFRSF7 Proteins and Enzymes
• CD27/TNFRSF7 ELISAs