title: Organotypic culture of HPV-transformed keratinocytes: a model for testing lymphocyte infiltration of (pre)neoplastic lesions of the uterine cervix.
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.
authors.affiliation: Department of Pathology B35, University Hospital of Liège, CHU Sart Tilman, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Jacobs
authors.forename: M P
authors.initials: MP
authors.lastname: Moutschen
authors.forename: E
authors.initials: E
authors.lastname: Franzen-Detrooz
authors.forename: V
authors.initials: V
authors.lastname: Boniver
authors.forename: J
authors.initials: J
authors.lastname: Boniver
authors.forename: P
authors.initials: P
authors.lastname: Delvenne
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis; D017252:CD3 Complex / Q000276:immunology; D019002:CD56 Antigen / Q000032:analysis; D002461:Cell Line, Transformed; D018920:Coculture Techniques; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007376:Interleukin-2 / Q000494:pharmacology; D015603:Keratinocytes / Q000276:immunology* / Q000378:metabolism; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology; D007694:Killer Cells, Natural / Q000187:drug effects / Q000502:physiology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000502:physiology*; D016246:Lymphocytes, Tumor-Infiltrating / Q000187:drug effects / Q000502:physiology*; D027383:Papillomaviridae; D010835:Phytohemagglutinins / Q000494:pharmacology; D017452:Receptors, IgG / Q000032:analysis; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D002583:Uterine Cervical Neoplasms / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D019002:CD56 Antigen; D007376:Interleukin-2; D010835:Phytohemagglutinins; D017452:Receptors, IgG; D018799:Intercellular Adhesion Molecule-1