ABSTRACT
A stable packaging cell line (Vero/BC-F) constitutively expressing fusion (F) protein of the human parainfluenza virus type 2 (hPIV2) was established for production of the F-defective and single round-infectious hPIV2 vector in a strategy for recombinant vaccine development. The F gene expression has not evoked cytostatic or cytotoxic effects on the Vero/BC-F cells and the F protein was physiologically active to induce syncytial formation with giant polykaryocytes when transfected with a plasmid expressing hPIV2 hemagglutinin-neuraminidase (HN). Transduction of the F-defective replicon RNA into the Vero/BC-F cells led to the release of the infectious particles that packaged the replicon RNA (named as hPIV2ΔF) without detectable mutations, limiting the infectivity to a single round. The maximal titer of the hPIV2ΔF was 6.0 × 10(8) median tissue culture infections dose per ml. The influenza A virus M2 gene was inserted into hPIV2ΔF, and the M2 protein was found to be highly expressed in a human lung cancer cell line after transduction. Furthermore, in vivo airway infection experiments revealed that the hPIV2ΔF was capable of delivering transgenes to hamster tracheal cells. Thus, non-transmissible or single round-infectious hPIV2 vector will be potentially applicable to human gene therapy or recombinant vaccine development.
Subject(s)
Gene Transfer Techniques , Genetic Vectors , Parainfluenza Virus 2, Human/genetics , Viral Fusion Proteins/genetics , Animals , Cell Line , Cells, Cultured , Chlorocebus aethiops , Cricetinae , Humans , Influenza A virus/genetics , Recombinant Proteins/genetics , Vaccines, Synthetic/genetics , Vero Cells , Viral Fusion Proteins/metabolism , Viral Matrix Proteins/geneticsABSTRACT
BACKGROUND: A new subline of the senescence accelerated mouse (SAM) P1/Yit strain has been established which shows spontaneous enteric inflammation under specific pathogen free (SPF) conditions. AIMS: To elucidate the pathogenesis of enteric inflammation in this new subline. METHODS: The SPF and germ free (GF) SAMP1/Yit strains were used. Histological, immunological, and microbiological characterisation of the mice with enteric inflammation was performed. RESULTS: Histologically, enteritic inflammation developed as a discontinuous lesion in the terminal ileum and caecum with the infiltration of many inflammatory cells after 10 weeks of age. the activity of myeloperoxidase, and both immunolocalisation and mRNA expression of inducible nitric oxide synthase increased in the lesion. CD3-epsilon positive T cells, neutrophils, and macrophages were more numerous in the inflamed mucosa of the SAMP1/Yit strain. The GF SAMP1/Yit strain did not show any inflammation in the intestinal wall, by the age of 30 weeks, and the enteritis and caecitis developed 10 weeks after the conventionalisation of the GF SAMP1/Yit strain. CONCLUSION: Enteric inflammation in the ileum and caecum developed in the SAMP1/Yit strain. The pathophysiological characteristics of the disease in this mouse have some similarities to those of human inflammatory bowel disease (IBD). This mouse strain should be a useful model system for elucidating the interaction between the pathogenesis of IBD and the gut microflora.
Subject(s)
Disease Models, Animal , Enteritis/etiology , Germ-Free Life , Inflammatory Bowel Diseases , Aging/metabolism , Animals , CD3 Complex , Cecal Diseases/metabolism , Cecal Diseases/microbiology , Cecum/metabolism , Cecum/microbiology , Enteritis/metabolism , Enteritis/microbiology , Granulocytes/immunology , Ileitis/metabolism , Ileitis/microbiology , Ileum/metabolism , Ileum/microbiology , Immunohistochemistry , Macrophages/immunology , Mice , Mice, Inbred AKR , Mice, Inbred Strains , Nitric Oxide Synthase/analysis , Nitric Oxide Synthase Type II , Peroxidase/analysisABSTRACT
A retrospective cohort study was carried out on asbestos workers who had received health examinations in 1972 to 1974 conducted by the Osaka Health Center. The subjects, total of 789 (329 males, 460 females) were followed-up for 10 years (Jan. 1, 1975-Dec. 31, 1984). There were sixty-one deaths in the cohort--4 tuberculosis, 12 malignant neoplasms (4 stomach cancers, 8 respiratory cancers including one case of pleural mesothelioma), 18 circulatory diseases, 24 respiratory diseases, and 3 other causes of death. Standardized mortality ratio (SMR) was calculated age and sex-specific death rates for the general population in Osaka between 1975-79 and 1980-84. SMR for all causes of death, stomach cancer, respiratory cancer, circulatory diseases, and respiratory diseases were 1.15, 3.29, 0.75, 3.88, 0.93 and 8.63 respectively. Respiratory cancer and respiratory diseases showed statistically significant (p less than 0.01) excess death with a mean death age of 59 and 56 years old respectively.