期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 331, 期 1-2, 页码 13-26出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2007.09.006
关键词
T cells; cloning; TCR; vbeta; tumor; human
资金
- NCI NIH HHS [R01-CA85843, CA16672, R01 CA085843-05, P01 CA049639-150021, P01-CA49639, 1K23 CA123149-01A1, K23 CA123149, P01 CA049639, P30 CA016672-32S3, K23 CA123149-01A1, P30 CA016672-32, R01 CA085843, P30 CA016672] Funding Source: Medline
Adoptive therapy with antigen-specific T cells is a promising approach for the treatment of infectious diseases and cancer. However, cloning of antigen-specific T cells by the traditional approach of limiting dilution is a time-consuming, laborious, and inefficient process. Here, we describe a novel flow cytometric strategy for rapid isolation of human tumor antigen-specific T-cell clones by using T-cell receptor (TCR) V beta antibodies in combination with carboxyfluorescein succinimidyl ester (CFSE)based proliferation assay. The USE dilution following antigen stimulation identified proliferating antigen-specific T cells, and the TCRV beta antibodies allowed distinguishing T cells at the clonal level from a heterogeneous T-cell population. This method of TCRV beta/CFSE dilution was used for the isolation of four different human lymphoma and melanoma-specific CD4(+) and CD8(+) T-cell clones reactive against defined and undefined tumor antigens. Isolated tumor-specific T-cell clones could be expanded to large numbers ex. vivo while maintaining phenotype, function, and tumor antigen specificity. The method was simple, efficient, and reproducible, and may have potential application for the development of adoptive immunotherapeutic strategies. (C) 2007 Elsevier B.V. All rights reserved.
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