Potential therapeutic strategy for non-Hodgkin lymphoma by anti-CD20scFvFc/CD28/CD3zeta gene tranfected T cells.
BACKGROUND
Anti-CD20 monoclonal antibody treatment has not only increased survival and cure rates in many non-Hodgkin lymphomas, but also has prompted an explosion in the development of novel antibodies and biologically active substances with specific cellular targets in the field of malignancies treatment. Since the robust immune responses are elicited by the gene-modified T cells, gene based T cell therapy may also provide a powerful tool for cancer immunotherapy.


METHODS
In this study, we developed a vector construction encoding a chimeric T cell receptor that recognizes the CD20 antigen and delivers co-stimulatory signals to achieve T cell activation. One non-Hodgkin lymphoma cell line Raji cells co-cultured with peripheral blood-derived T cells were stably transfected with anti-CD20scFvFc/CD28/CD3zeta gene or anti-CD20scFvFc gene. T cells expressing anti-CD20scFvFc/CD28/CD3zeta or anti-CD20scFvFc gene co-cultured with CD20 positive Raji cells for different times. Cell lysis assay was carried by [3H]TdR release assay. The expressions of Fas, Bcl-2 and Caspase-3 of Raji cells were detected by flow cytometric. The secretion of IFN-gamma and IL-2 in co-culture medium was tested by ELISA assay. Activity of AP-1 was analyzed by EMSA.


RESULTS
Following efficient transduction of peripheral blood-derived T cells with anti-CD20scFvFc/CD28/CD3zeta gene, an obvious cell lysis of Raji cells was observed in co-culture. T cells transduced anti-CD20scFvFc/CD28/CD3zeta gene had superior secretion of IFN-gamma and IL-2 compared to T cells transduced anti-CD20scFvFc gene. Also it led to a much stronger Fas-induced apoptosis signaling transduction in target cancer cells.


CONCLUSIONS
So adoptively T cells transduced anti-CD20scFvFc/CD28/CD3zeta gene mediates enhanced anti-tumor activities against CD20 positive tumor cells, suggesting a potential of gene-based immunotherapy for non-Hodgkin lymphoma.
Zheng
Yihu
Y
Institute of Medical Research, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou, China.
Yu
Kang
K
Du
Jimei
J
Jiang
Lei
L
Zhang
Shenghui
S
Han
Yixiang
Y
Yu
Panpan
P
Tan
Yingxia
Y
D000911:Antibodies, Monoclonal / Q000235:genetics*; D018951:Antigens, CD20 / Q000276:immunology*; D018106:CD28 Antigens / Q000235:genetics*; D017252:CD3 Complex / Q000235:genetics*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D048430:Cell Shape; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation*; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014162:Transfection; D019014:fas Receptor / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D018106:CD28 Antigens; D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; C491901:FAS protein, human; C508594:IL2 protein, human; D007376:Interleukin-2; D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; D018808:Transcription Factor AP-1; D019014:fas Receptor; D007371:Interferon-gamma; C505407:CASP3 protein, human; D053148:Caspase 3
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