title: Simultaneous immunisation with a Wilms' tumour 1 epitope and its ubiquitin fusions results in enhanced cell mediated immunity and tumour rejection in C57BL/6 mice.
abstract: Protein fusion to ubiquitin results in its targeting to proteasome and processing through MHC class I pathway. We used this approach to induce cytotoxic T lymphocyte (CTL) response against a MHC class I epitope. Therefore, two known proteasome targeting systems, "ubiquitin fusion degradation" (UFD) and "N-end rule", were used to immunise C57BL/6 mice. Two plasmids encoding an epitope from Wilms' Tumour 1 (WT1-126), fused N-terminally to ubiquitin, were constructed. They were designated as "pUbVVPT" and "pUbGRPT", targeting the fused epitope to UFD and N-end pathways, respectively. A plasmid encoding WT1-126 without ubiquitin fusion (pPT) was also constructed as control. Three mice groups were immunised using these constructs (UGR, UVV and PT groups). Two other groups received mixed immunisations of pUbVVPT or pUbGRPT plus pPT plasmids (UVV+PT and UGR+PT). All mice received a WT1-126 peptide booster. Lymphoproliferative responses following stimulation with WT1-126 were observed in all immunisation groups, with mice receiving the mixture of plasmids eliciting the highest proliferation (UVV+PT>UGR+PT>PT). Moreover, In vivo cytotoxicity assay results revealed highest specific lysis of target cells in UVV+PT group. Tumour growth was decreased in all immunised groups, and was completely abrogated in UGR+PT group. In addition, T(H)1 type cytokines patterns were detected from all immunised groups and WT1-126-specific IFNγ producing lymphocytes were developed in them. These results suggest that the delivery of ubiquitin-fused epitopes along with epitopes alone can be used to optimise the effect of DNA vaccines on the induction of anti-tumour immunity.
authors.affiliation: Department of Immunology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Nasir Saeedi
authors.initials: NS
authors.lastname: Eslami
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Shokrgozar
authors.forename: Asadollah
authors.initials: A
authors.lastname: Mousavi
authors.forename: Kayhan
authors.initials: K
authors.lastname: Azadmanesh
authors.forename: Alireza
authors.initials: A
authors.lastname: Nomani
authors.forename: Vasso
authors.initials: V
authors.lastname: Apostolopoulos
authors.forename: Stephanie
authors.initials: S
authors.lastname: Day
authors.forename: Amir
authors.initials: A
authors.lastname: Amanzadeh
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Alimohammadian
mesh_terms: D000818:Animals; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D016207:Cytokines / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005260:Female; D005805:Genes, MHC Class I / Q000235:genetics / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D007111:Immunity, Cellular / Q000235:genetics / Q000276:immunology; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D025801:Ubiquitin / Q000235:genetics / Q000276:immunology*; D054875:Ubiquitination; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology; D025721:WT1 Proteins / Q000235:genetics / Q000276:immunology*; D009396:Wilms Tumor / Q000235:genetics / Q000276:immunology / Q000628:therapy*
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; D019444:Vaccines, DNA; D025721:WT1 Proteins; D007371:Interferon-gamma; D046988:Proteasome Endopeptidase Complex