Anti-CD22 (HA22SH) h(ICOS-4-1BB-CD3ζ) CAR, pSBCAR1 vector

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

Sleeping Beauty (SB) transposon, a type of nonviral integrative vectors, provides an alternative to modify primary T cells. Creative biolabs has developed SB transposon CAR vector pSBCAR1 CD22 (HA22SH) h(ICOSBBζ), which is constructed for the engineering of T cells to target human CD22. The T cells are genetically modified through transduction with a nonviral vector expressing scFv of anti-CD22 antibody linked to ICOS (CD278) signaling domain and CD137 (4-1BB), CD3-zeta signaling domains. And the vector product was designed for the treatment of Acute lymphoblastic leukemia (ALL).

Details

Target
CD22

Targeting Cell Type
T cell

Targeting Diseases
Acute lymphoblastic leukemia (ALL)

Generation
Third

Vector Name
pSBCAR1

Vector Length
~6kb

Vector Type
Sleeping Beauty (SB) transposon

Receptor Construction
scFv-ICOS-4-1BB-CD3ζ

Discription of Signaling Cassetes
ICOS
CD278 or ICOS (Inducible T-cell COStimulator) is a CD28-superfamily costimulatory molecule that is expressed on activated T cells. It is thought to be important for Th2 cells in particular. The protein encoded by this gene belongs to the CD28 and CTLA-4 cell-surface receptor family. It forms homodimers and plays an important role in cell-cell signaling, immune responses, and regulation of cell proliferation.
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 antigen-nonspecific 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 ICOS and 4-1BB 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 FuncS

Fig.1 Comparison of CD22-CAR and CD19-CAR-mediated lysis.

CAR Construction : HA22SH-CD28-CD3ζ

Fig.1 Comparison of CD22-CAR and CD19-CAR-mediated lysis.

51Cr-release assay to evaluate lytic activity of CD22 HA22SH-28z second-generation CAR (inverted triangle), m971-28z second-generation CAR (gray triangle), CD19 CAR (squares), or mock transduced T cells (circles) against ALL lines.

Haso, W., Lee, D. W., Shah, N. N., Stetler-Stevenson, M., Yuan, C. M., Pastan, I. H., ... & Orentas, R. J. (2013). Anti-CD22–chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood, The Journal of the American Society of Hematology, 121(7), 1165-1174.

Complete CAR data AM

Fig.2 Cytokine release by CAR-transduced T cells.

CAR Construction : HA22SH-CD28-CD3ζ, HA22SH-CD28-41BB-CD3ζ

Fig.2 Cytokine release by CAR-transduced T cells.

CAR-transduced T cells (vectors listed, x-axis) were incubated with irradiated CD22-high (Raji, column 1), CD22-low (NALM6-GL, column 2), or CD22-negative (K562, column 3) leukemia cell lines at a ratio of 10:1 for 24 hours and culture supernatants analyzed for IFN-γ (top row), IL-2 (second row), and TNF-α (third row).

Complete CAR data BI

Fig.3 Evaluation of CD22 CARs in vivo.

CAR Construction : HA22SH-CD28-CD3ζ

Fig.3 Evaluation of CD22 CARs in vivo.

On day 0, NSG mice were injected intravenously with 5 × 10^5 NALM6-GL cells. On day 3 mice received 1 × 10^7 CAR+ T cells (HA22SH-28z (n = 5) or m971-28z (n = 5) or mock T cells (n = 5). Bioluminescent imaging pre-treatment (day 3), day 7 and day 15 after intravenous injection of NALM6-GL.

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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.