Anti-CEA (A5B7) 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

CAR scFv data ELISA

Fig.1 Anti CEA3 anti-CD3 bispecific diabody-mediated T-cell cytotoxicity against the CEA-expressing cell lines H498.

CAR Construction :

Fig.1 Anti CEA3 anti-CD3 bispecific diabody-mediated T-cell cytotoxicity against the CEA-expressing cell lines H498.

Values are displayed as the mean of triplicate measurements.

Holliger, P., Manzke, O., Span, M., Hawkins, R., Fleischmann, B., Qinghua, L., ... & Bohlen, H. (1999). Carcinoembryonic antigen (CEA)-specific T-cell activation in colon carcinoma induced by anti-CD3× anti-CEA bispecific diabodies and B7× anti-CEA bispecific fusion proteins. Cancer Research, 59(12), 2909-2916.

CAR scFv data FCM

Fig.2 Colon carcinoma tissue specimens were analyzed for CEA expression by flow cytometry.

CAR Construction :

Fig.2 Colon carcinoma tissue specimens were analyzed for CEA expression by flow cytometry.

All measurements were performed in triplicate.

CAR scFv data FCM

Fig.3 T-cell stimulation in primary colon carcinoma. After 5 days, the cells were harvested and analyzed by flow cytometry for differences in the numbers of CD4- and CD8-positive T cells, and the T cell:tumor cell ratio was calculated from the flow cytometric data.

CAR Construction :

All measurements were performed in triplicate.

CAR scFv data FCM

Fig.4 Effect of incubation time of IFN-g on CEA-expression demonstrated with the A5B7 anti-CEA monoclonal antibody, analysed flow cytometrically.

CAR Construction :

Fig.4 Effect of incubation time of IFN-g on CEA-expression demonstrated with the A5B7 anti-CEA monoclonal antibody, analysed flow cytometrically.

Mean fluorescence intensities are given in the upper right corner.

Verhaar, M. J., Damen, C. A., Zonnenberg, B. A., & Blijham, G. H. (1999). In vitro upregulation of carcinoembryonic antigen expression by combinations of cytokines. Cancer letters, 139(1), 67-73.

CAR scFv data FCM

Fig.5 The effect of IFN-g as a single agent and in combination with IL-6 on CEA-expression on HT29-19A tumour cells.

CAR Construction :

Fig.5 The effect of IFN-g as a single agent and in combination with IL-6 on CEA-expression on HT29-19A tumour cells.

All incubation times are
48 h. Mean ¯uorescence intensities are given in the upper right corner.

Verhaar, M. J., Damen, C. A., Zonnenberg, B. A., & Blijham, G. H. (1999). In vitro upregulation of carcinoembryonic antigen expression by combinations of cytokines. Cancer letters, 139(1), 67-73.

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 (A5B7) h(CD28-OX40-CD3ζ) CAR, pSBCAR1 (CAR-SB-02LX362)

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