Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes

F. Grünebach, M. R. Müller, A. Nencioni, P. Brossart

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Dendritic cells (DC) are professional antigen-presenting cells playing a central role in the induction of antigen-specific cytotoxic T-lymphocytes (CTL). We analyzed the efficiency of tumor RNA transfection into DC using different sources of RNA as well as delivery strategies including electroporation, lipofection and CD71-receptor-based delivery. To evaluate the sensitivity of these approaches, we utilized in vitro transcribed enhanced green fluorescence protein (EGFP)-RNA and whole tumor RNA from EGFP-transfected renal cell carcinoma cell line N43. We demonstrate that electroporation was the most effective way yielding about 30% EGFP positive cells while less than 1% of DC expressed EGFP using the transferrin receptor transfection system. Delivery of RNA with liposomes resulted in 17.5% of EGFP positive cells depending on the RNA amount. However, when these approaches were applied to transduce DC with RNA derived from the A498 cell line for T-cell priming, tumor-specific CTL could be induced using all delivery strategies suggesting that this technology has the potential to induce cytotoxic T-cell response even when low level of antigen is delivered. Furthermore, we demonstrate that amplification of whole tumor messenger RNA (mRNA) as well as the use of total instead of purified mRNA can be utilized for stimulating tumor-specific CTL responses.

Original languageEnglish
Pages (from-to)367-374
Number of pages8
JournalGene Therapy
Volume10
Issue number5
DOIs
Publication statusPublished - Mar 2003

Fingerprint

Cytotoxic T-Lymphocytes
RNA
Fluorescence
Dendritic Cells
Neoplasms
Electroporation
Proteins
Transfection
T-Lymphocytes
Antigens
Cell Line
Messenger RNA
Transferrin Receptors
Antigen-Presenting Cells
Renal Cell Carcinoma
Liposomes
Technology

Keywords

  • CTL
  • Dendritic cells
  • Immunotherapy
  • RNA transfection

ASJC Scopus subject areas

  • Genetics

Cite this

Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes. / Grünebach, F.; Müller, M. R.; Nencioni, A.; Brossart, P.

In: Gene Therapy, Vol. 10, No. 5, 03.2003, p. 367-374.

Research output: Contribution to journalArticle

Grünebach, F. ; Müller, M. R. ; Nencioni, A. ; Brossart, P. / Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes. In: Gene Therapy. 2003 ; Vol. 10, No. 5. pp. 367-374.
@article{4ff0b4dc8f2d420ca562c7cb5f5c11ca,
title = "Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes",
abstract = "Dendritic cells (DC) are professional antigen-presenting cells playing a central role in the induction of antigen-specific cytotoxic T-lymphocytes (CTL). We analyzed the efficiency of tumor RNA transfection into DC using different sources of RNA as well as delivery strategies including electroporation, lipofection and CD71-receptor-based delivery. To evaluate the sensitivity of these approaches, we utilized in vitro transcribed enhanced green fluorescence protein (EGFP)-RNA and whole tumor RNA from EGFP-transfected renal cell carcinoma cell line N43. We demonstrate that electroporation was the most effective way yielding about 30{\%} EGFP positive cells while less than 1{\%} of DC expressed EGFP using the transferrin receptor transfection system. Delivery of RNA with liposomes resulted in 17.5{\%} of EGFP positive cells depending on the RNA amount. However, when these approaches were applied to transduce DC with RNA derived from the A498 cell line for T-cell priming, tumor-specific CTL could be induced using all delivery strategies suggesting that this technology has the potential to induce cytotoxic T-cell response even when low level of antigen is delivered. Furthermore, we demonstrate that amplification of whole tumor messenger RNA (mRNA) as well as the use of total instead of purified mRNA can be utilized for stimulating tumor-specific CTL responses.",
keywords = "CTL, Dendritic cells, Immunotherapy, RNA transfection",
author = "F. Gr{\"u}nebach and M{\"u}ller, {M. R.} and A. Nencioni and P. Brossart",
year = "2003",
month = "3",
doi = "10.1038/sj.gt.3301901",
language = "English",
volume = "10",
pages = "367--374",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes

AU - Grünebach, F.

AU - Müller, M. R.

AU - Nencioni, A.

AU - Brossart, P.

PY - 2003/3

Y1 - 2003/3

N2 - Dendritic cells (DC) are professional antigen-presenting cells playing a central role in the induction of antigen-specific cytotoxic T-lymphocytes (CTL). We analyzed the efficiency of tumor RNA transfection into DC using different sources of RNA as well as delivery strategies including electroporation, lipofection and CD71-receptor-based delivery. To evaluate the sensitivity of these approaches, we utilized in vitro transcribed enhanced green fluorescence protein (EGFP)-RNA and whole tumor RNA from EGFP-transfected renal cell carcinoma cell line N43. We demonstrate that electroporation was the most effective way yielding about 30% EGFP positive cells while less than 1% of DC expressed EGFP using the transferrin receptor transfection system. Delivery of RNA with liposomes resulted in 17.5% of EGFP positive cells depending on the RNA amount. However, when these approaches were applied to transduce DC with RNA derived from the A498 cell line for T-cell priming, tumor-specific CTL could be induced using all delivery strategies suggesting that this technology has the potential to induce cytotoxic T-cell response even when low level of antigen is delivered. Furthermore, we demonstrate that amplification of whole tumor messenger RNA (mRNA) as well as the use of total instead of purified mRNA can be utilized for stimulating tumor-specific CTL responses.

AB - Dendritic cells (DC) are professional antigen-presenting cells playing a central role in the induction of antigen-specific cytotoxic T-lymphocytes (CTL). We analyzed the efficiency of tumor RNA transfection into DC using different sources of RNA as well as delivery strategies including electroporation, lipofection and CD71-receptor-based delivery. To evaluate the sensitivity of these approaches, we utilized in vitro transcribed enhanced green fluorescence protein (EGFP)-RNA and whole tumor RNA from EGFP-transfected renal cell carcinoma cell line N43. We demonstrate that electroporation was the most effective way yielding about 30% EGFP positive cells while less than 1% of DC expressed EGFP using the transferrin receptor transfection system. Delivery of RNA with liposomes resulted in 17.5% of EGFP positive cells depending on the RNA amount. However, when these approaches were applied to transduce DC with RNA derived from the A498 cell line for T-cell priming, tumor-specific CTL could be induced using all delivery strategies suggesting that this technology has the potential to induce cytotoxic T-cell response even when low level of antigen is delivered. Furthermore, we demonstrate that amplification of whole tumor messenger RNA (mRNA) as well as the use of total instead of purified mRNA can be utilized for stimulating tumor-specific CTL responses.

KW - CTL

KW - Dendritic cells

KW - Immunotherapy

KW - RNA transfection

UR - http://www.scopus.com/inward/record.url?scp=0037344753&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037344753&partnerID=8YFLogxK

U2 - 10.1038/sj.gt.3301901

DO - 10.1038/sj.gt.3301901

M3 - Article

VL - 10

SP - 367

EP - 374

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 5

ER -