Construction of Anti-CD20 Single-Chain Antibody-CD28-CD137-TCRζ Recombinant Genetic Modified T Cells and its Treatment Effect on B Cell Lymphoma

• Published in: Medical science monitor 2015-07, Vol.21, p.2110-2115
• Main Authors: Chen, Fei, Fan, Chuming, Gu, Xuezhong, Zhang, Haixi, Liu, Qian, Gao, Xiaoli, Lu, Jie, He, Baoli, Lai, Xun
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD - genetics; Antigens, CD - immunology; Antigens, CD20 - genetics; Antigens, CD20 - immunology; CD28 Antigens - genetics; CD28 Antigens - immunology; CD4-Positive T-Lymphocytes - immunology; Gene Fusion; HEK293 Cells; Humans; Immunotherapy, Adoptive - methods; Lab/In Vitro Research; Lymphoma, B-Cell - therapy; Mice; Mice, Nude; Random Allocation; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Recombination, Genetic; Single-Chain Antibodies - genetics; Single-Chain Antibodies - immunology; Transfection - methods; Tumor Necrosis Factor Receptor Superfamily, Member 9 - genetics; Tumor Necrosis Factor Receptor Superfamily, Member 9 - immunology
• ISSN: 1643-3750; 1234-1010; 1643-3750
• DOI: 10.12659/MSM.893791

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. 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. 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. 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.