title: Construction of Anti-CD20 Single-Chain Antibody-CD28-CD137-TCRζ Recombinant Genetic Modified T Cells and its Treatment Effect on B Cell Lymphoma.
abstract: 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.
  
  
  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.
authors.affiliation: Department of Nephrology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Fei
authors.initials: F
authors.lastname: Chen
authors.affiliation: Intensive Care Unit, Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Chuming
authors.initials: C
authors.lastname: Fan
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Xuezhong
authors.initials: X
authors.lastname: Gu
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Haixi
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Gao
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.affiliation: Animal Laboratory, Kunming Medical University, Kunming, Yunnan, China (mainland).
authors.forename: Baoli
authors.initials: B
authors.lastname: He
authors.affiliation: Department of Hematology, The First People's Hospital of Yunnan Province, Kunming, Yunnan, China (mainland).
authors.forename: Xun
authors.initials: X
authors.lastname: Lai
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000276:immunology; D018951:Antigens, CD20 / Q000235:genetics / Q000276:immunology; D018106:CD28 Antigens / Q000235:genetics / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D050939:Gene Fusion; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D016393:Lymphoma, B-Cell / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D011897:Random Allocation; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D011995:Recombination, Genetic; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology; D014162:Transfection / Q000379:methods*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics / Q000276:immunology
chemical_list: D015703:Antigens, CD; D018951:Antigens, CD20; D018106:CD28 Antigens; D011948:Receptors, Antigen, T-Cell; D057127:Single-Chain Antibodies; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; C054979:antigen T cell receptor, zeta chain