Anti-CEA (MFZ) h(CD28-OX40-CD3ζ) CAR, pSBCAR1 vector

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

Details

Target
CEA

Targeting Cell Type
T cell

Targeting Diseases
Pancreatic cancer

Generation
Third

Vector Name
pSBCAR1

Vector Length
~6kb

Vector Type
Sleeping Beauty (SB) transposon

Receptor Construction
scFv-CD28-OX40-CD3ζ

Discription of Signaling Cassetes
CD28
CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatorysignals 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.
OX40
OX40, also known as CD134 or TNFRSF4 is a member of the TNFR-superfamily of receptors which is not constitutively expressed on resting naïve T cells, unlike CD28. OX40 is a secondary co-stimulatory immune checkpoint molecule, expressed after 24 to 72 hours following activation. The interaction between OX40 and its binding partner, OX40L (CD252) plays an important role in antigen-specific T-cell expansion and survival. OX40 and OX40L also regulate cytokine production from T cells and modulate cytokine receptor signaling. OX40 cosignaling in CAR improve redirected T-cell effector functions and enhance anti-tumor activity.
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 FCM

Fig.1 Both CD4 and CD8 T cells were modified with the CAR and used for adoptive transfer in this study

CAR Construction : MFZ scFv-28ζ

Fig.1 Both CD4 and CD8 T cells were modified with the CAR and used for adoptive transfer in this study

CAR expression was monitored by flow cytometry using a phycoerythrin-labeled anti-mouse IgG1 antibody and an allophycocyanin-conjugated antibody directed against CD4 and CD8.

Chmielewski, M., Hahn, O., Rappl, G., Nowak, M., Schmidt–Wolf, I. H., Hombach, A. A., & Abken, H. (2012). T cells that target carcinoembryonic antigen eradicate orthotopic pancreatic carcinomas without inducing autoimmune colitis in mice. Gastroenterology, 143(4), 1095-1107.

Complete CAR data Cyt

Fig.2 T cells with anti-CEA CAR were coincubated at the indicated effector-to-tumor cell ratios with Panc02 cells with or without CEA expression for 48 hours.

CAR Construction : MFZ scFv-28ζ

Fig.2 T cells with anti-CEA CAR were coincubated at the indicated effector-to-tumor cell ratios with Panc02 cells with or without CEA expression for 48 hours.

Cytotoxicity was assessed by an XTT-based viability assay. All measurements were performed in triplicate.

Complete CAR data FCM

Fig.3 Lung epithelia cells were isolated from the lung of CEAtg mice. Cells were stained for CEA and flow sorted before use in the study.

CAR Construction : MFZ scFv-28ζ

Fig.3 Lung epithelia cells were isolated from the lung of CEAtg mice. Cells were stained for CEA and flow sorted before use in the study.

Mouse CEA Panc02 pancreatic carcinoma cells and fibrosarcoma CEA C15A3 cells were used for comparison.

Complete CAR data IHC

Fig.4 CEA Panc02 pancreatic carcinoma cells were inoculated into the pancreas of CEAtg mice. After 3 weeks, the tumor tissue, as well as the healthy pancreas (without tumor), were recorded by H&E and CEA staining.

CAR Construction : MFZ scFv-28ζ

The CEAtg mouse expressed CEA in transplanted CEA+ Panc02 pancreatic carcinoma as well as in healthy tissues.

Complete CAR data FuncS

Fig.5 Regression of transplanted pancreas carcinoma upon adoptive therapy with anti-CEA CAR T cells.

CAR Construction : MFZ scFv-28ζ

Fig.5 Regression of transplanted pancreas carcinoma upon adoptive therapy with anti-CEA CAR T cells.

CBLuc-marked Panc02 tumor cells and GLuc-marked T cells were recorded by bioluminescence imaging in the same mouse at the indicated days before and after T-cell transfer.

More Published Data More Published Data

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.

Anti-CEA (MFZ) h(CD28-OX40-CD3ζ) CAR, pSBCAR1 (CAR-SB-02LX358)

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