title: DNA alkylating agents alleviate silencing of class II transactivator gene expression in L1210 lymphoma cells.
abstract: MHC class II (Ia) Ag expression is inversely correlated with tumorigenicity and directly correlated with immunogenicity in clones of the mouse L1210 lymphoma (1 ). Understanding the mechanisms by which class II Ag expression is regulated in L1210 lymphoma may facilitate the development of immunotherapeutic approaches for the treatment of some types of lymphoma and leukemia. This study demonstrates that the variation in MHC class II Ag expression among clones of L1210 lymphoma is due to differences in the expression of the class II transactivator (CIITA). Analysis of stable hybrids suggests that CIITA expression is repressed by a dominant mechanism in class II-negative L1210 clones. DNA-alkylating agents such as ethyl methanesulfonate and the chemotherapeutic drug melphalan activate CIITA and class II expression in class II negative L1210 cells, and this effect appears to be restricted to transformed cell lines derived from the early stages of B cell ontogeny. Transient transfection assays demonstrated that the CIITA type III promoter is active in class II(-) L1210 cells, despite the fact that the endogenous gene is not expressed, which suggests that these cells have all of the transacting factors necessary for CIITA transcription. An inverse correlation between methylation of the CIITA transcriptional regulatory region and CIITA expression was observed among L1210 clones. Furthermore, 5-azacytidine treatment activated CIITA expression in class II-negative L1210 cells. Collectively, our results suggest that 1) CIITA gene expression is repressed in class II(-) L1210 cells by methylation of the CIITA upstream regulatory region, and 2) treatment with DNA-alkylating agents overcomes methylation-based silencing of the CIITA gene in L1210 cells.
authors.affiliation: Department of Immunology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA. shawn.murphy@roswellpark.org
authors.forename: Shawn P
authors.initials: SP
authors.lastname: Murphy
authors.forename: Renae
authors.initials: R
authors.lastname: Holtz
authors.forename: Nicole
authors.initials: N
authors.lastname: Lewandowski
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Tomasi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Fuji
mesh_terms: D020121:5' Untranslated Regions / Q000187:drug effects; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D002999:Clone Cells; D019175:DNA Methylation / Q000187:drug effects; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects* / Q000276:immunology*; D020868:Gene Silencing / Q000187:drug effects*; D005802:Genes, MHC Class II / Q000187:drug effects*; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis; D006825:Hybridomas; D007939:Leukemia L1210 / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008811:Mice, Inbred DBA; D009687:Nuclear Proteins; D011401:Promoter Regions, Genetic / Q000187:drug effects; D015534:Trans-Activators / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D020121:5' Untranslated Regions; D018906:Antineoplastic Agents, Alkylating; D000949:Histocompatibility Antigens Class II; C083728:MHC class II transactivator protein; D009687:Nuclear Proteins; D015534:Trans-Activators; D005020:Ethyl Methanesulfonate