bromochloroacetic-acid and Retroviridae-Infections

A mutant of MoMuLV, called ts1, causes an AIDS-like syndrome in susceptible strains of mice. In mice infected at birth, thymic atrophy, CD4+ T cell loss, body wasting, and death occur by approximately 30-40 days postinfection (dpi). We have shown previously that the death of ts1-infected cells is not caused by viral replication per se, but by oxidative stress and apoptosis following their accumulation the ts1 viral envelope precursor protein, gPr80(env). In infected mice treated with the antioxidant monosodium alpha-luminol (GVT), T cell loss and thymic atrophy are delayed for many weeks, and body wasting and death do not occur until long after infected, untreated control mice have died. We show here that GVT treatment of ts1-infected mice maintains the thymic epithelial cell (TEC) cytoarchitecture and cytokeratin gradients required for thymocyte differentiation. It also suppresses thymocyte reactive oxygen species (ROS) levels, upregulates and stabilizes levels of the antioxidant-regulating transcription factor Nrf2, and prevents accumulation of gPr80(env) in thymocytes. We conclude that GVT treatment can make ts1 a non-cytopathic virus for thymocytes, although it cannot prevent thymocyte infection. Since oxidative stress also contributes to the loss of T cells in HIV-AIDS, the antioxidant effects of GVT may make it a useful therapeutic adjunct to HAART treatment.

Topics: Animals; Antioxidants; CD4-Positive T-Lymphocytes; Cell Survival; Cytopathogenic Effect, Viral; Epithelial Cells; Gene Expression Regulation, Viral; Immune Tolerance; Keratins; Leukemia Virus, Murine; Leukemia, Experimental; Mice; Mutation; NF-E2-Related Factor 2; Oxidative Stress; Retroviridae Infections; Retroviridae Proteins; Thymus Gland

To establish an oral epithelial cell line of mouse origin for molecular and biochemical assays.. Epithelial cells were isolated from the oral cavity of adult mice and established as a spontaneously immortalized cell line in culture, designated immortalized oral keratinocyte cells (IMOK cells). The cells were then characterized for growth characteristics, differentiation potential, karyotype, transfectability, susceptibility to viral infection and responses to siRNA.. The IMOK cells exhibit robust growth in both serum-containing and serum-free medium for at least 100 population doublings. IMOK cells have a near diploid karyotype, express keratinocyte marker proteins and can be induced to undergo differentiation by the addition of high levels of calcium to the medium. The differentiation process is characterized by morphological changes and by the induction of oral epithelium specific differentiation marker proteins such as K4 and K13. Transient transfection analyses reveal that IMOK cells are highly transfectable and that several promoters of epithelial cells are active in these cells. Moreover, upon differentiation with calcium, there is an up-regulation of differentiation-specific K4 and Elf5 promoter activity. Finally, we show that the oral keratinocytes are also amenable to infection with retroviruses and to siRNA-based knockdown of gene expression.. Our study is the first to establish an immortalized oral keratinocyte cell line of murine origin that can recapitulate the oral epithelium differentiation program and thus could serve as a useful tool for toxicological and molecular analyses of the oral tissue.

Topics: Animals; Calcium; Cell Culture Techniques; Cell Differentiation; Cell Line, Transformed; Culture Media; Epithelial Cells; Female; Gene Expression; Gene Knockdown Techniques; Karyotyping; Keratinocytes; Keratins; Mice; Mouth Mucosa; Promoter Regions, Genetic; Retroviridae Infections; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; Transfection