ABSTRACT
AIM: Overexpression of Wilms' tumour gene (WT1) has been proven in several tumours. Previous research of our group on the cell cycle of uterine leiomyosarcoma (LMS) and carcinosarcoma (CS) suggested a possible role for WT1. We therefore intended to further explore the expression pattern of WT1 in uterine sarcomas. METHODS: 27 CS, 38 LMS, 15 endometrial stromal sarcomas (ESS) and seven undifferentiated sarcomas (US) were collected. WT1 expression was evaluated by immunohistochemistry (IHC) in 87 samples, by RT-PCR (m-RNA expression) in 23 random selected samples and by Western blotting in 12 samples, separating cytoplasmic and nuclear proteins. A pilot study to detect mutations (exons 7-10) was performed on eight samples. RESULTS: IHC showed WT1 positivity in 12/27 CS, 29/38 LMS, 7/15 ESS and 4/7 US. All-but-one sample had a positive RT-PCR. All Western blottings were positive with more cytoplasmic expression in 9/12 cases. No mutations were found. CONCLUSIONS: WT1 is overexpressed in uterine sarcomas. Since increased levels of mRNA determine the biological role, WT1 might contribute to uterine sarcoma tumour biology.
Subject(s)
Genes, Wilms Tumor , Mutation/genetics , Sarcoma/genetics , Uterine Neoplasms/genetics , Blotting, Western , DNA Mutational Analysis , DNA, Neoplasm/analysis , Female , Humans , Immunohistochemistry , Reverse Transcriptase Polymerase Chain Reaction , Up-RegulationABSTRACT
Plasmid DNA (pcDNA1::MOMP D) expressing the major outer membrane protein (MOMP) of an avian Chlamydophila psittaci serovar D strain was tested for its ability to induce protective immunity against C. psittaci challenge in the presence of maternal antibodies. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was used. Following pcDNA1::MOMP vaccination, both T helper and B cell memory were primed. However, high maternal antibodies titres affected the induction of vaccine-specific antibody responses as assessed by MOMP-specific antibody levels in enzyme-linked immunosorbent assay (ELISA). Cell-mediated immunity was unaltered as demonstrated by the significantly heightened proliferative responses of peripheral blood lymphocytes (PBL) following vaccination. DNA vaccination could significantly reduce clinical symptoms, pharyngeal and cloacal excretion as well as Chlamydophila replication, even in the presence of maternal antibodies.
Subject(s)
Antibodies, Bacterial/biosynthesis , Chlamydophila psittaci/immunology , DNA, Bacterial/immunology , Turkeys/immunology , Administration, Intranasal , Animals , Antibodies, Bacterial/analysis , Bacterial Outer Membrane Proteins/immunology , Chlamydophila psittaci/isolation & purification , Eggs , Enzyme-Linked Immunosorbent Assay , Female , Fluorescent Antibody Technique , Injections, Intramuscular , Lymphocytes/immunology , Psittacosis/immunology , Psittacosis/pathology , Respiratory System/microbiology , Vaccination , Vaccines, DNA/immunologyABSTRACT
Plasmid DNA (pcDNA1::MOMP D) expressing the major outer membrane protein of an avian Chlamydophila psittaci serovar D strain was tested for its ability to induce protective immunity against Chlamydophila psittaci challenge in the presence of maternal antibodies. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was used. Only placebo-vaccinated turkeys showed a primary response following challenge, although DNA vaccination didn't generate high antibody titres. Following pcDNA::MOMP vaccination, both T-helper and B-cell memory were primed. However, high maternal antibodies titres affected the induction of vaccine-specific antibody responses as assessed by MOMP-specific antibody levels in ELISA. Cell-mediated immunity was unaltered as demonstrated by the significantly heightened proliferative responses of peripheral blood lymphocytes following vaccination. DNA vaccination could significantly reduce clinical symptoms, pharyngeal and cloacal excretion as well as chlamydophila replication, even in the presence of maternal antibodies.
