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Rapid induction of cytotoxic T-cell response against cervical cancer cells by human papillomavirus type 16 E6 antigen gene delivery into human dendritic cells by an adeno-associated virus vector

Cancer Gene Therapy volume 8, pages 948–957 (2001)Cite this article

Abstract

We have shown that the pulsing of dendritic cells (DCs) with human papillomavirus type 16 (HPV-16) antigen proteins by lipofection stimulates class I–restricted cytotoxic T lymphocyte (CTL) response against primary cervical cancer cells. Also, we have shown that adeno-associated virus (AAV) was able to effectively deliver a cytokine gene into DCs. It has been our hypothesis that the delivery of antigen genes into DCs, resulting in endogenous and continuous antigen protein expression, may result in an improvement in T-cell priming by DCs. Here, DCs are pulsed (infected) with an AAV vector containing the HPV-16 E6 gene. After infection, transduced E6 gene mRNA expression and vector chromosomal integration could be identified in infected DCs. Furthermore, priming rosettes formed at early times when the AAV/E6 vector was used. Most importantly, AAV/E6 vector pulsing of DCs induced, after only 7 days of priming, a strong CTL response against primary cervical cancer cell lines, compared to bacterial E6 protein lipofection. Killing was significantly blocked by the addition of anti-MHC class I antibodies. Fluorescence-activated cell sorter (FACS) analysis of resulting primed cell populations revealed higher levels of CD8+ T cells by AAV-based pulsing, with little evidence of CD56 (NK). FACS analysis of the DC populations revealed that AAV/E6 vector–pulsed DCs had higher levels of CD80 and lower levels of CD86 than protein-pulsed DCs. These data suggest that rAAV may be appropriate for antigen pulsing of DCs for immunotherapy protocols. Finally, our protocol represents an advance in regards to the time needed for generating a CTL response compared to other techniques. Cancer Gene Therapy (2001) 8, 948–957

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Acknowledgements

Drs. Yong Liu and Maurizio Chiriva-Internati contributed equally to this work. We acknowledge the excellent assistance provided by Teri Fields and Rena Sheffer.

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Authors and Affiliations

  1. Departments of Obstetrics and Gynecology, University of Arkansas for Medical Sciences, Little Rock, 72205, Arkansas, USA

    Yong Liu, Juan J Roman & Paul L Hermonat

  2. Center for Immunology and Microbial Disease, Albany Medical College, Albany, 12208, New York, USA

    Maurizio Chiriva-Internati, Emanuela Salati & Seah Lim

  3. Scientific Direction, Instituto Clinico, Humanitas, Rozzano, Milano, Italy

    Fabio Grizzi

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  1. Yong Liu
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  2. Maurizio Chiriva-Internati
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  3. Fabio Grizzi
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  7. Paul L Hermonat
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Correspondence to Paul L Hermonat.

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Liu, Y., Chiriva-Internati, M., Grizzi, F. et al. Rapid induction of cytotoxic T-cell response against cervical cancer cells by human papillomavirus type 16 E6 antigen gene delivery into human dendritic cells by an adeno-associated virus vector. Cancer Gene Ther 8, 948–957 (2001). https://doi.org/10.1038/sj.cgt.7700391

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