Related Experiment Videos
Characterization of antigen-presenting properties of tumour cells using virus-specific cytotoxic T lymphocytes
D C Spierings1, E Agsteribbe, J Wilschut
1Department of Medical Microbiology, University of Groningen, The Netherlands.
Abstract:
Immunotherapy of tumours by induction of tumour-specific cytotoxic T-lymphocytes (CTLs) will only be effective for tumours with a functional antigen processing and presentation machinery. However, many tumours are known to down-regulate expression of major histocompatibility complex (MHC) class I molecules and/or to impair antigen processing. It is therefore desirable to evaluate the ability of a given tumour to present antigenic epitopes before developing an immunotherapy protocol. In this study we have used influenza virus as a tool to determine the antigen-presenting capacities of the murine neuroblastoma C1300 cell line NB41A3, a frequently used model for human neuroblastoma. Immunofluorescence analyses revealed low and moderate expression of MHC class I molecules Dd and Kk respectively. Nevertheless, infected NB41 A3 cells were lysed efficiently by influenza-specific CTLs. These results demonstrate that all steps of the antigen-processing pathway function properly in the NB tumour cells, and that the limited MHC class I expression suffices for efficient recognition by CTLs. In addition, lysis of the NB tumour cells shows that the cells are susceptible to CTL-induced apoptosis, a pathway that is often impaired in tumour cells. These characteristics make neuroblastoma a suitable target for immunotherapy. The presented assay allows evaluation of various immunological properties of tumour cells and, thus, represents a valuable tool to assess whether a given tumour will be susceptible to immunotherapy or not.
Insights
Neuroblastoma cells can be effectively targeted by immunotherapy. This study shows that even with low major histocompatibility complex (MHC) class I expression, neuroblastoma cells can present antigens and are susceptible to T-cell mediated killing.
Area of Science:
- Immunology
- Oncology
- Virology
Background:
- Immunotherapy efficacy relies on tumor antigen processing and presentation.
- Many tumors downregulate major histocompatibility complex (MHC) class I or impair antigen processing.
- Assessing tumor antigen-presenting capacity is crucial before immunotherapy development.
Purpose of the Study:
- To evaluate the antigen-presenting capacity of murine neuroblastoma C1300 cell line (NB41A3) using influenza virus.
- To determine if neuroblastoma cells are susceptible to T-cell mediated apoptosis.
Main Methods:
- Immunofluorescence analysis of MHC class I (Dd and Kk) expression on NB41A3 cells.
- Assessment of cytotoxic T-lymphocyte (CTL) mediated lysis of influenza-infected NB41A3 cells.
Main Results:
- NB41A3 cells exhibited low and moderate expression of MHC class I molecules Dd and Kk.
- Influenza-infected NB41A3 cells were efficiently lysed by influenza-specific CTLs.
- Neuroblastoma cells demonstrated susceptibility to CTL-induced apoptosis.
Conclusions:
- Neuroblastoma cells possess a functional antigen processing and presentation machinery.
- Limited MHC class I expression is sufficient for effective CTL recognition.
- Neuroblastoma is a suitable target for immunotherapy, and the developed assay can predict treatment susceptibility.