Anti-EphA2 (XHM268) h(CD28-CD3ζ) CAR, pCDCAR1 vector

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Details

Target
EphA2

Targeting Cell Type
T cell

Targeting Diseases
Breast cancer, Ovarian cancer, Prostate cancer

Generation
Second

Vector Name
pCDCAR1

Vector Length
~8kb

Vector Type
Lentiviral vector

Receptor Construction
scFv-CD28-CD3ζ

Discription of Signaling Cassetes
CD28
CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide costimulatory signals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells (APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR+ T cells containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy.
CD3ζ
CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ , ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta, which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

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Published Data

Complete CAR data ELISA

Fig.1 Erythropoietin-producing hepatocellular carcinoma A2 (EphA2)-specific T cells recognize and kill EphA2-positive gliomas.

CAR Construction : 4H5 scFv-41BB-CD28ζ

Fig.1 Erythropoietin-producing hepatocellular carcinoma A2 (EphA2)-specific T cells recognize and kill EphA2-positive gliomas.

Nontransduced (NT) or EphA2-specific T cells were cocultured with target cells at a 1:5 ratio, and after 24 hours, the production of interferon-γ (IFN-γ) and interleukin- 2 (IL-2) by T cells was determined by ELISA.

Chow, K. K., Naik, S., Kakarla, S., Brawley, V. S., Shaffer, D. R., Yi, Z., Rainusso, N., Wu, M. F., Liu, H., Kew, Y., Grossman, R. G., Powell, S., Lee, D., Ahmed, N., & Gottschalk, S. (2013). T cells redirected to EphA2 for the immunotherapy of glioblastoma. Molecular therapy : the journal of the American Society of Gene Therapy, 21(3), 629–637.

Complete CAR data Cyt

Fig.2 Cytotoxicity assays.

CAR Construction : 4H5 scFv-41BB-CD28ζ

Fig.2 Cytotoxicity assays.

Cytotoxicity assays with EphA2-specific T cells and NT-T cells as effectors and five primary EphA2-positive glioblastoma cell lines as targets.

Complete CAR data BI

Fig.3 Bioluminescence imaging.

CAR Construction : 4H5 scFv-41BB-CD28ζ

Fig.3 Bioluminescence imaging.

Bioluminescence imaging was used to follow tumor progression. All mice had detectable tumors just prior to treatment (day 7). By day 40, all mice in the control groups had progressive tumors (9/9 for untreated, 8/8 for NT-T cells). The tumor signal decreased in all mice treated with EphA2-specific T cells, and 50% (6/12) were long-term survivors.

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.