21 July 2015 : Laboratory Research
Construction of Anti-CD20 Single-Chain Antibody-CD28-CD137-TCRζ Recombinant Genetic Modified T Cells and its Treatment Effect on B Cell LymphomaFei ChenBC, Chuming FanCD, Xuezhong GuDE, Haixi ZhangEF, Qian LiuFG, Xiaoli GaoDF, Jie LuCG, Baoli HeFG, Xun LaiABCD
Med Sci Monit 2015; 21:2110-2115
BACKGROUND: Immunotherapy has been explored as a new therapy for B cell lymphoma, which is a non-Hodgkin’s lymphoma. Because CD20 is a B lymphocyte-specific marker, anti-CD20 single chain-tagged T lymphocytes have already begun to be experimentally used in B cell lymphoma treatment, but its use is still limited because of its unspecific targeting. T cells transfected with CD28 and CD137 can significantly improve the ability of cytokines secretion and anti-tumor effect, as well as extending T cell survival time and improving their proliferation ability.
MATERIAL AND METHODS: Genes containing anti-CD20-CD28-CD137-TCRζ were constructed. After cloning and sequencing, the plasmid was constructed and packaged by lentivirus. It was transfected to the peripheral blood T lymphocyte after identification transfection to induce the fusion protein expression. The cells were incubated with Raji cells and the LDH test was performed to detect the cytotoxic effect of CAR-T cells; the tumor volume and survival rate were measured to observe its inhibitory effect on B cell lymphoma in nude mice.
RESULTS: Gene with anti-CD20-CD28-CD137-TCRζ was successfully constructed and transfected to the T cell surface. LDH assay revealed that CAR-T cells can kill the Raji cells with a killing rate of 32.89±6.26%. It can significantly inhibit B cell lymphoma growth in nude mice.
CONCLUSIONS: T lymphocytes transfected with anti-CD20-CD28-CD137-TCRζ fusion gene can kill B cell lymphoma, which could provide a new strategy for tumor treatment.
Keywords: Antigens, CD - immunology, Animals, Antigens, CD137 - immunology, Antigens, CD20 - immunology, Antigens, CD28 - immunology, CD4-Positive T-Lymphocytes - immunology, Gene Fusion, HEK293 Cells, Immunotherapy, Adoptive - methods, Lymphoma, B-Cell - therapy, Mice, Mice, Nude, Random Allocation, Receptors, Antigen, T-Cell - immunology, Recombination, Genetic, Single-Chain Antibodies - immunology, Transfection - methods
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