T cells harboring anti-CD20 scFv-CD8-TCR? fusion gene in treatment of human B-cell lymphomas: an experimental study
- VernacularTitle:抗CD20单链抗体-CD8-TCR?融合基因转染的T细胞治疗人B细胞淋巴瘤的实验研究
- Author:
Lin YUN
- Publication Type:Journal Article
- Keywords:
anti-CD20 scFv-CD8-TCR? fusion gene;
T-lymphocytes;
lymphoma,B-cell
- From:
Academic Journal of Second Military Medical University
2000;0(07):-
- CountryChina
- Language:Chinese
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Abstract:
Objective:To observe the anti-tumor effect of T lymphocytes harboring anti-CD20 scFv-CD8-TCR? fusion gene on human B lymphomas in vitro and in vivo,so as to explore the feasibility of CD20-mediated autogenous T lymphocytes in killing B-cell lymphomas.Methods: A fusion gene containing anti-CD20 scFv,CD8 molecule and CD3? chain was constructed and was cloned into pcDNA3.After confirmed by restriction endonuclease analysis,the fusion gene was used to transfect the human peripheral T lymphocytes through electroporation and expression of anti-CD20 scFv-CD8-TCR? fusion protein was induced.The CD20 antigen-recognition ability of transfected T cells was determined by flow cytometry.The killing effect of transfected T cells on B cell lymphomas-Raji cells was tested in a cytotoxicity assay.The antitumor efficacy of this gene-modified T cell against the Raji tumor cell line was also evaluated in BALB/c nude mice.Results: We successfully constructed the anti-CD20 scFv-CD8-TCR? fusion gene and expressed its protein on T cells.Flow cytometry and cytotoxicity assay demonstrated that the gene engineered T cells specifically recognized CD20 antigen and specifically inhibited B-cell lymphomas Raji cells.Furthermore,the T cells significantly inhibited the transplanted Raji cells in irradiated BALB/c nude mice.Conclusion: T lymphocytes transfected with anti-CD20 scFv-CD8-TCR? fusion gene have antigen-specific anti-tumor effect in vitro and in vivo,which lays a foundation for utilizing human T lymphocytes to treat human B-cell lymphomas.
