Premium
Expression of mucin (MUC‐1) from a Mini‐Epstein‐Barr virus in immortalized B‐cells to generate tumor antigen specific cytotoxic T cells
Author(s) -
Kilger Ellen,
Pecher Gabriele,
Schwenk Arvid,
Hammerschmidt Wolfgang
Publication year - 1999
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/(sici)1521-2254(199903/04)1:2<84::aid-jgm21>3.0.co;2-q
Subject(s) - cytotoxic t cell , mucin , antigen , virology , epstein–barr virus , virus , biology , tumor cells , cancer research , immunology , in vitro , genetics , biochemistry
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. Copyright © 1999 John Wiley & Sons, Ltd.