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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M.D. Henryk Pilch
Department of Obstetrics and Gynecology
Johannes Gutenberg University Mainz
D-55101 Mainz