Cervical Cancer Cells Present HPV-E7 Epitope to CD4+ Tumor Infiltrating Lymphocytes (TIL) in Association with Histocompatibility Leukocyte Antigen (HLA) DR4

Abstract

Purpose: Cellular and humoral immune responses directed against autologous tumor cells in patients with cervical cancer may be directed against proteins provided by human papilloma- virus (HPV) associated products or, alternatively, by yet undefined targets as well. The goal of our study was to evaluate the local cellular immune response in cervical cancer. Methods: From a fresh tumor sample of a patient with cervical carcinoma, tumor-infiltrating lymphocytes (TIL) were generated, expanded and characterized by immunohistochemistry, flow cytometry, cytokine release assays and DNA fragment analysis. Results: We established a MHC class II-restricted CD4+ T-cell line from a patient with cervical cancer which recognizes autologous (HPV35+, HPV59+) tumor cells and the HLA-DR4-matched cervical cancer cell line Me180 (HPV68+) as determined by TNFα secretion. Expression of different HPV-E7 genes in autologous B-cells revealed that this T-cell line defines a DR4-presented T-cell epitope which is shared among the E7 gene of HPV59 and HPV68. Conclusion: Tumor-HPV-specific and MHC-class II-restricted CD4+ T-cells are present within the tumor lesion and can be successfully expanded in the presence of IL-2 and IL-7. MHC class II presented peptides may be implemented to augment T-cell responses directed against autologous tumor cells.

Zusammenfassung

Fragestellung: Zelluläre und humorale Immunantworten gegen autologe Tumorzellen von Patientinnen mit Gebärmutterhalskrebs können gegen humanes Papillomavirus (HPV) assoziierte Epitope oder aber gegen andere, noch nicht definierte Tumor-assoziierte Zielstrukturen gerichtet sein. Das Ziel dieser Untersuchung bestand darin, die lokale intratumorale Immunantwort im Falle eines Zervixkarzinoms zu untersuchen. Material und Methoden: Aus nativem Tumorgewebe einer Patientin mit einem fortgeschrittenen Zervixkarzinom wurden tumorinfiltrierende Lymphozyten (TIL) generiert, expandiert und mittels Immunhistochemie, Flowzytometrie, Zytokin-Freisetzungstest sowie DNA-Fragmentanalyse charakterisiert. Ergebnisse: Es gelang uns, eine MHC-Klasse-II-restringierte CD4+ T-Zell-Linie aus dem Tumor einer Patientin mit Gebärmutterhalskarzinom zu etablieren, die gemessen an einer TNFαSekretion autologe (HPV35+, HPV59+) Tumorzellen sowie Zellen der HLA-DR4 positiven Zervixkarzinomzelllinie Me180 (HPV68+) erkennt. Die Expression verschiedener transfizierter HPV-E7Gene in autologen B-Zellen lässt erkennen, daß diese T-Zelllinie ein MHC-Klasse-II präsentiertes T-Zellepitop definiert, welches den E7-Genen von HPV59 und HPV68 offenbar gemeinsam ist.Schlußfolgerung: Wir konnten zeigen, daß Tumor/HPV-spezifische und MHC-Klasse-II-restringierte CD4+ T-Zellen in der Tumorläsion zu detektieren sind, die durch IL-2/IL-7 erfolgreich expandiert werden können. MHC-Klasse-II-präsentierte Proteine könnten durchaus eingesetzt werden, um eine T-zellvermittelte Immuntherapie gegen autologe Zervixkarzinomzellen zu induzieren bzw. zu steigern.

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