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| Item Type: | Article |
|---|---|
| Title: | Expression of mucin (MUC-1) from a mini-Epstein-Barr virus in immortalized B-cells to generate tumor antigen specific cytotoxic T cells |
| Creators Name: | Kilger, E., Pecher, G., Schwenk, A. and Hammerschmidt, W. |
| Abstract: | Background: EBV immortalized B-cells can be used as antigen presenting cells (APC) to stimulate specific T-cell responses. Mini-Epstein-Barr virus (mini-EBV) plasmids contain all functional elements of Epstein-Barr virus (EBV) necessary to immortalize B-cells in vitro. These immortalized B-cells are incapable of releasing infectious virus in contrast to cells immortalized by wildtype EBV. In addition, mini-EBVs can be modified in E. coli to alter their genetic composition or adopt new genes. Methods: We constructed a mini-EBV plasmid carrying an expression cassette for the human tumor antigen mucin encoded by the gene MUC-1. Primary human B-cells were infected with the MUC-1 carrying mini-EBV plasmid packaged into an EBV coat and immortalized B-cell clones were expanded in vitro. These B-cells were analyzed by FACS analyses for the expression of mucin and co-stimulatory molecules and were subsequently used as antigen presenting cells (APC) to stimulate peripheral blood mononuclear cells from healthy donors. Results: Several B-cell lines were established that were shown to be free of helper virus or wildtype EBV. These B-cells expressed the relevant tumor-specific epitopes of mucin and the co-stimulatory ligands B7.1 and B7.2 necessary for efficient T-cell activation. Using the mucin expressing B-cells as antigen presenting cells (APC) mucin-epitope specific cytotoxic T-cells were established. Conclusions: Virus-free B-cell lines expressing tumor-associated epitopes such as mucin or other antigens of interest provide an unlimited and safe source of APC to generate antigen specific T-cells which could be used for clinical trials in adoptive immune therapy or cancer vaccines. |
| Keywords: | Antigen Presenting Cells, EBV, MUC-1 |
| Source: | Journal of Gene Medicine |
| ISSN: | 1099-498X |
| Publisher: | John Wiley & Sons Ltd |
| Volume: | 1 |
| Number: | 2 |
| Page Range: | 84-92 |
| Date: | March 1999 |
| Official Publication: | https://doi.org/10.1002/(SICI)1521-2254(199903/04)1:2<84::AID-JGM21>3.0.CO;2-Q |
| PubMed: | View item in PubMed |
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