Uncoordinate induction and differential regulation of HLA class-I and class-II expression by gamma-interferon in differentiating human neuroblastoma cells.

Abstract

Recombinant gamma-interferon (IFN-gamma) has recently been shown to be one of the most effective inducers of neuroblastoma (NB) cell differentiation. Since increasing evidence indicates that expression of MHC class-I and class-II antigens by tumour cells is important for immunorecognition and cell targeting, we tested whether induction of NB cell differentiation by IFN-gamma is followed by expression of HLA class-I and class-II molecules. LAN-5 human NB cell line completely lacks HLA class-I antigens. Their expression was induced in a dose-dependent manner by IFN-gamma. HLA class-II molecules are also absent on LAN-5 cells, but only DP antigens were dose-dependently induced by IFN-gamma, while DR and DQ molecules were unaffected by the treatment. To confirm and extend the immunological data to all the class-II molecules, we performed Northern blot analysis, observing that DP alpha mRNA was induced in a dose- and time-dependent manner. DO beta and DZ alpha genes were also induced peaking after 3 days of IFN-gamma treatment. DR beta and DQ beta genes, which were not induced by IFN-gamma, gave a normal pattern of enzyme restriction by Southern blot. To get an insight into the regulation of HLA class-II gene expression in the neuronal model, we measured the decline of the steady-state HLA class-II mRNA. DO beta mRNA rapidly returned to baseline level after removing IFN-gamma, while the decay rates of DP alpha and DZ alpha mRNA were very slow. This might indicate different regulation at the post-transcriptional level for DO beta mRNA with respect to DP alpha and DZ alpha mRNA. To strengthen these findings we evaluated the half-lives of the mRNA after IFN-gamma induction by means of actinomycin D treatment. HLA-DO beta mRNA had a shorter half-life, while DZ alpha and DP alpha had a longer decay rate. Finally, we report that treatment of LAN-5 cells with cycloheximide did not alter the rate of transcription of the HLA-DP alpha gene, suggesting that no protein factor(s) is/are needed to maintain DP alpha gene expression.