Anti-CEA (MFZ) h(41BB-CD3ζ) CAR, pMMLV 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
Second

Vector Name
pMMLV

Vector Length
~6kb

Vector Type
Recombinant Moloney murine leukemia virus (MMLV) retroviral vector

Receptor Construction
scFv-41BB-CD3ζ

Discription of Signaling Cassetes
41BB
CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigennonspecific chimeric antigen receptor T-cells to significantly increases antitumor 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 4-1BB 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(41BB-CD3ζ) CAR, pMMLV (CAR-MV-01LX364)

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