Radons, Juergen and Gross, Catharina and Stangl, Stefan and Multhoff, Gabriele (2005) Nucleofection of non-B cells with mini-Epstein-Barr virus DNA. JOURNAL OF IMMUNOLOGICAL METHODS, 303 (1-2). pp. 135-141. ISSN 0022-1759, 1872-7905
Full text not available from this repository. (Request a copy)Abstract
A tumor-specific cell surface localization of heat shock protein 70 (Hsp70) on CX+ colon carcinoma cells provides a recognition structure for NK cells but not for NKT and T cells. Incubation with low-dose IL-2 plus Hsp70-peptide TKD enhances production and release of granzyme B by NK cells and thus renders Hsp70-positive tumors more sensitive to their cytolytic attack. To provide the experimental basis for the generation of Hsp70-reactive NK cell lines we established a modified nucleofection technique as a rapid and efficient method for gene transfer into non-B cells. Therefore, TKD-stimulated, CD3/CD19-depleted effector cells, consisting of 85% CD3(-) CD16/56(+) NK cells, 1.4% CD3(+) CD16/56+ NKT cells, and 0.3% CD3(+) CD16/56(-) T cells were nucleofected with the green fluorescent protein (GFP)-containing mini-Epstein-Barr virus (mini-EBV) plasmid p2667 (1478.A d2GFP). GFP, a marker for the expression of EBV-associated genes, became visible for the first time on day 18 after transfection. On day 28 mini-EBV-transfected cells consisted of 49% NKT, 38% T cells, and 13% NK cells; no contaminating B cells were detectable. Even 1.5 years after transfection GFP and CD94 were found to be co-expressed on transfectants. These data indicated that mini-EBV provides a useful tool for the nucleofection of non-B cells. The cytolytic activity of NK-transfectants towards Hsp70 membrane-positive CX+ tumor cells was comparable to that of non-transfected effector cells. In summary, our results might provide the basis for the generation of non-B effector cell lines including NK cells with conserved Hsp70-reactivity. (c) 2005 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | NATURAL-KILLER-CELLS; CYTOTOXIC T-CELLS; CYTOLYTIC ACTIVITY; BOUND HEAT-SHOCK-PROTEIN-70; RECOGNITION STRUCTURE; GRANZYME-B; NK CELLS; CD94; non-B cells; NK cells; nucleofection; Mini-EBV; granzyme B |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 03 May 2021 08:28 |
| Last Modified: | 03 May 2021 08:28 |
| URI: | https://pred.uni-regensburg.de/id/eprint/35821 |
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