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Anti-CD5 (H65) h(CD28-CD3ζ) CAR, pCDCAR1 (CAR-LC173)


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

The vector of anti-CD5 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD5. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-CD5 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of acute myeloid leukemia.

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Details

  • Target
  • CD5
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Acute Lymphoblastic Leukemia
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Retroviral
  • 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 co-stimulatory 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

  • Clone
  • H65
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • CD5
  • Synonyms
  • CD5; T1; LEU1

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  • Published Data
Complete CAR data FCM

Fig.1 Cytotoxicity assay in vitro.

CAR Construction : H65-CD28-CD3ζ Latest CAR Construction

Fig.1 Cytotoxicity assay in vitro.

Cytotoxicity of CD19 CAR- and CD5 CAR-transduced T cells against T-ALL and T-lymphoma cell lines was assessed in a 5-hour Cr release assay.

Mamonkin, M., Rouce, R. H., Tashiro, H., & Brenner, M. K. (2015). A T-cell–directed chimeric antigen receptor for the selective treatment of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 126(8), 983-992.

Complete CAR data ELISA

Fig.2 Determination of cytokines release.

CAR Construction : H65-CD28-CD3ζ Latest CAR Construction

Fig.2 Determination of cytokines release.

Production of IFN-γ and TNF-α by CD4+(top) and CD8+(bottom) T cells transduced with CD19 CAR or CD5 CAR was measured by intracellular cytokine staining.

Mamonkin, M., Rouce, R. H., Tashiro, H., & Brenner, M. K. (2015). A T-cell–directed chimeric antigen receptor for the selective treatment of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 126(8), 983-992.

Complete CAR data BI

Fig.3 CD5 CAR T cells control progression of T-ALL in xenograft mouse models.

CAR Construction : H65-CD28-CD3ζ Latest CAR Construction

Fig.3 CD5 CAR T cells control progression of T-ALL in xenograft mouse models.

Jurkat-FFluc cells (Fig.A-E) or CCRF-CEM-FFluc cells (Fig.F-H) were IV injected followed by IV injection of CAR T cells on days 3 and 6 postimplantation (Fig.A, F) or on day 6 and 9 post-implantation (Fig.C).

Mamonkin, M., Rouce, R. H., Tashiro, H., & Brenner, M. K. (2015). A T-cell–directed chimeric antigen receptor for the selective treatment of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 126(8), 983-992.

Complete CAR data FCM

Fig.4 The immunophenotype of CD5CAR T cells.

CAR Construction : H65-41BB-CD3ζ Latest CAR Construction

Fig.4 The immunophenotype of CD5CAR T cells.

The majority of CD5KO anti-CD5 CAR-T cells displayed a naive-like surface phenotype that might have an enhanced capacity for expansion, differentiation, and self-renewal upon antigen stimulation.

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

Complete CAR data Cyt

Fig.5 Cytotoxicity assay in vitro.

CAR Construction : H65-41BB-CD3ζ Latest CAR Construction

Fig.5 Cytotoxicity assay in vitro.

Anti-CD5 CAR-T cells exhibited cytotoxicity in vitro when co-incubated with malignant T cell lines with moderate expression of CD5 antigen at relatively low effector to target (E:T) ratios

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

Complete CAR data FuncS

Fig.6 Cytokine secretion and repeated stimulation assay of CD5-targeting CAR-T cells in vitro.

CAR Construction : H65-41BB-CD3ζ Latest CAR Construction

Fig.6 Cytokine secretion and repeated stimulation assay of CD5-targeting CAR-T cells in vitro.

CAR-T cells produced the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and interferon (IFN)-γ and elevated levels of interleukin-2 (IL-2) release when co-incubated with CCRF-CEM cells.

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

Complete CAR data BI

Fig.7 The in vivo antitumor activity of CD5-targeting CAR-T cells in the tumor model established by CCRF-CEM.

CAR Construction : H65-41BB-CD3ζ Latest CAR Construction

Fig.7 The in vivo antitumor activity of CD5-targeting CAR-T cells in the tumor model established by CCRF-CEM.

A mouse tumor model of T cell acute lymphoblastic leukemia (T-ALL) by tail intravenous injection of CCRF-CEM-ffLuc cells was established to verify the in vivo efficacy of CD5 CAR-T cells.

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

Complete CAR data BI

Fig.8 The in vivo antitumor activity of CD5-targeting CAR-T cells in the tumor model established by SUP-T1.

CAR Construction : H65-41BB-CD3ζ Latest CAR Construction

Fig.8 The in vivo antitumor activity of CD5-targeting CAR-T cells in the tumor model established by SUP-T1.

A xenograft mouse tumor model was established by SUP-T1 cells with moderate CD5 expression to verify the in vivo efficacy of CD5 CAR-T cells.

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

CAR scFv data SPR

Fig.9 The binding affinity measurement of H65 antibody to CD5 antigen.

CAR Construction : Latest CAR Construction

Fig.9 The binding affinity measurement of H65 antibody to CD5 antigen.

The affinity of H65 to recombinant human CD5 was determined using bio-layer interferometry.

Dai, Z., Mu, W., Zhao, Y., Jia, X., Liu, J., Wei, Q., ... & Zhou, J. (2021). The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains. Molecular Therapy, 29(9), 2707-2722.

More Published Data More Published Data

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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.

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