Virchows Archiv

, Volume 432, Issue 4, pp 323–330

Organotypic culture of HPV-transformed keratinocytes: a model for testing lymphocyte infiltration of (pre)neoplastic lesions of the uterine cervix

  • N. Jacobs
  • Michel P. Moutschen
  • Elisabeth Franzen-Detrooz
  • Vincent Boniver
  • Jacques Boniver
  • Philippe Delvenne
ORIGINAL ARTICLE

DOI: 10.1007/s004280050173

Cite this article as:
Jacobs, N., Moutschen, M., Franzen-Detrooz, E. et al. Virchows Archiv (1998) 432: 323. doi:10.1007/s004280050173

Abstract

 The aim of our study was to establish the relevance of an in vitro model for analysing the ability of human lymphocytes to infiltrate human papillomavirus (HPV)-associated (pre)neoplastic lesions of the uterine cervix. To mimic these lesions, we have used the organotypic raft culture of HPV-transformed keratinocytes (SiHa). The SiHa organotypic raft culture was co-cultured with resting or prestimulated (IL-2 or IL-2+anti-CD3 mAb) allogeneic peripheral blood mononuclear cells (PBMC) for 24 and 72 h. The majority of infiltrating cells were T lymphocytes. Occasional NK cells were also identified. The stimulation with IL-2+anti-CD3 mAb induced the highest number of infiltrating cells, with the maximum lymphocyte infiltration observed after 24 h of co-culture. The lymphocyte infiltration was associated with an increased number of apoptotic cells in the organotypic cultures. The ability of PBMC and purified T cell and NK cell populations to lyse HPV-transformed keratinocytes was also investigated on monolayer cultures. As expected in an allogenic model, the highest cytotoxicity was mediated by NK cells activated by IL-2 or IL-2+anti-CD3 mAb. The cytotoxic activity of T cells was weak but, interestingly, increased in the presence of phytohaemagglutinin A (PHA), assuming that T cells were able to kill HPV-infected keratinocytes when a bridge between T cells and keratinocytes was provided. In conclusion, the organotypic culture of HPV-transformed keratinocytes may provide an effective in vitro model for investigating novel T cell-based immunotherapy protocols for the treatment of HPV-associated lesions.

Key words Human papillomavirus Cervical neoplastic lesions Organotypic culture T lymphocytes LAK cells 

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • N. Jacobs
    • 1
  • Michel P. Moutschen
    • 1
  • Elisabeth Franzen-Detrooz
    • 1
  • Vincent Boniver
    • 1
  • Jacques Boniver
    • 1
  • Philippe Delvenne
    • 1
  1. 1.Department of Pathology B35, University Hospital of Liège, CHU Sart Tilman, B-4000 Liège, Belgium Tel.: (+32) 43664282, Fax: (+32) 43662919 e-mail: N.Jacobs@ulg.ac.beBE

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