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The vector of anti-CD22 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD22. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD22 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of relapsed or refractory or MRD+ CD22+ B-cell acute lymphoblastic leukemia (B-ALL).
CAR Construction : m971-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. |
CAR Construction : m971-CD28-CD3ζ Fig.2 Flow cytometric analysis of CD22 expression on 7 primary patient pre-B ALL samples. lysis of patient blasts with m971-28z or HASH-28z-expressing T cells in a 4 hours 51Cr-release assay, percent lysis indicated at E/T ratio of 30:1. 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. |
CAR Construction : m971-CD28-CD3ζ, m971-CD28-41BB-CD3ζ Fig.3 Cytokine release by CAR-transduced T cells. CAR-transduced T cells were incubated with irradiated CD22-high (Raji), CD22-low (NALM6-GL), or CD22-negative (K562) leukemia cell lines at a ratio of 10:1 for 24 hours and culture supernatants analyzed for IFN-γ , IL-2, and TNF-α. 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. |
CAR Construction : m971-CD28-CD3ζ Fig.4 Evaluation of CD22 CARs in vivo (BI). Bioluminescent imaging pre-treatment (day 3), day 7 and day 15 after intravenous injection of NALM6-GL. 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. |
CAR Construction : m971-CD28-CD3ζ Fig.5 Evaluation of CD22 CARs in vivo (Kaplan-Meier survival curves). Kaplan-Meier survival curves for each group, listing significant differences between each group, survival statistics calculated using log-rank (Mantel-Cox) analysis. 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. |
CAR Construction : m871-CD28-41BB-CD3ζ Fig.6 Comparing the activation of RFB4 CAR with m971 CAR against the full-length CD22. RFB4 or m971 CAR-Jurkats were cocultured with Raji B cells (E:T=1:1) for 24 hrs. (B) IL-2 release was measured by ELISA and (C) CD69 expression was assessed by flow cytometry. Xiao, Q., Zhang, X., Tu, L., Cao, J., Hinrichs, C. S., & Su, X. (2022). Size-dependent activation of CAR-T cells. Science Immunology, 7(74), eabl3995. |
CAR Construction : Fig.7 Affinity determination of m971 and m972. Increasing concentrations of both IgG1 m971 and IgG1 m972 were incubated with BJAB cells in a flow cytometry assay. Dimitrov, D.S., Xiao, X.D., Pastan, I.H., (2013). Human monoclonal antibodies specific for CD22. US 8.591,889 B2, The United States of America as represented by the Secretary of the Department of Health and Human Services, Washington, DC (US). |
CAR Construction : Fig.8 Affinity determination of m971 and m972. The median fluorescence intensities from flow cytometry were plotted against the corresponding concentrations used for IgG1 m971 and IgG1 m972. Dimitrov, D.S., Xiao, X.D., Pastan, I.H., (2013). Human monoclonal antibodies specific for CD22. US 8.591,889 B2, The United States of America as represented by the Secretary of the Department of Health and Human Services, Washington, DC (US). |
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