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Archive > Volume 27 Issue 8 > 2020,27(8):852-859. DOI:10.3872/j.issn.1007-385X.2020.08.003 Prev Next
Basic Research
The killing activity of a CD7 nanobody derived CAR-T cells on CD7 positive acute myeloid leukemia cells
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Abstract:
Objective: To develop a new type of CD7 chimeric antigen receptor modified T cell (CD7-CAR-T) for the treatment of CD7 positive acute myeloid leukemia (AML), and to observe its killing effect on CD7 positive AML cells. Methods: The CD7-CAR lentiviral vector was constructed based on the CD7 Nanobody sequence and costimulatory domain sequence of CD28 and 4-1BB. The lentiviral particles were packaged and used to co-transfect human T cells with protein expression blocker (PEBL), so as to prepare CD7CAR-T cells. Real time cellular analysis (RTCA) was used to monitor the cytotoxicity of CD7-CAR-T cells on CD7 overexpressed 293T cells. Flow cytometry assay was used to detect the effect of CD7-CAR-T cells on proliferation and cytokine secretion of AML cells with high, medium and low CD7 expressions (KG-1, HEL and Kasumi-1 cells, respectively). Results: CD7-CAR-T cell was successfully constructed and its surface expression of CD7 was successfully blocked. Compared with T cells, CD7-CAR-T cells could significantly inhibit the proliferation of CD7-293T cells and promote the release of TNF, Granzyme B and INF-γ; in addition, CD7CAR-T cells also significantly promoted the apoptosis (t=147.1, P<0.01; t=23.57, P<0.01) and cytokine release (P<0.05 or P<0.01) in CD7 positive KG-1 and HEL cells, but had little effect on Kasumi-1 cells that only expressed minimal CD7 antigen (t=0.7058, P>0.05). Conclusion: CD7-CAR-T cells can specifically kill CD7-positiveAMLcells in vitro.
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Project Supported:
Project supported by the National Key R&D Program of China (No. 2016YFC1303403), the Natural Science Foundation of China (No. 81872431 and No. 31471283), the Priority Academic Program Development of Jiangsu Higher Education Institutions, the Collaborative Innovation Major Project (No. XYXT-2015304), the Six Talent Peaks Project in Jiangsu Province (No. SWYY-CXTD010), and the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No. 19KJD320003)
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