ABSTRACT
The present study describes the replication of Theiler's virus in cloned cerebrovascular endothelial cells (CVE) isolated from strains of mice that are either susceptible or resistant to Theiler's virus-induced demyelination (TVID). CVE isolated from all strains of mice were equally permissive to Theiler's virus infection. Interferon-gamma and tumor necrosis factor-alpha were found to inhibit the replication of Theiler's virus in CVE. A correlation between susceptibility to demyelination and the ability of Theiler's virus to induce MHC Class I on CVE was demonstrated.
Subject(s)
Brain/blood supply , Endothelium, Vascular/virology , Poliomyelitis/immunology , Theilovirus , Acute Disease , Animals , Brain/virology , Clone Cells , Endothelium, Vascular/immunology , Histocompatibility Antigens Class I/analysis , Histocompatibility Antigens Class II/analysis , Interferon-gamma/pharmacology , Mice , Mice, Inbred BALB C , Mice, Inbred CBA , Poliomyelitis/virology , T-Lymphocytes/physiology , Theilovirus/drug effects , Theilovirus/physiology , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Cloned cerebrovascular endothelial cells (CVE) persistently infected with Theiler's virus (PI-CVE) have been established and characterized. The CVE were derived from strains of mice that are susceptible (SJL/J and CBA) and resistant (BALB/c) to Theiler's virus-induced demyelination (TVID). The cells were persistently infected with either the BeAn or GDVII strains of Theiler's virus in vitro and studied at various passage levels for infectious virus, viral antigen and the expression of major histocompatibility complex (MHC) Class I and II antigens. The virus replicated to lower titers than in acutely infected CVE and appeared to be more cell-associated. Flow cytometric analysis revealed that 18-39% of the PI-CVE contained viral antigen. Persistently infected CVE derived from SJL/J and CBA mice expressed high levels of MHC Class I, whereas BALB/c PI-CVE did not. MHC Class II was upregulated by IFN-gamma in SJL/J PI-CVE albeit at a slightly lower level than in uninfected CVE. In addition, the PI-CVE demonstrated increased levels of mRNA for IL-1 beta when compared to uninfected CVE.
Subject(s)
Brain/blood supply , Endothelium, Vascular/virology , Poliomyelitis/immunology , Theilovirus , Animals , Antigens, Viral/analysis , Brain/virology , Clone Cells , Endothelium, Vascular/immunology , Flow Cytometry , Histocompatibility Antigens Class I/analysis , Histocompatibility Antigens Class II/analysis , Interleukin-1/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred CBA , Poliomyelitis/virology , Theilovirus/immunology , Theilovirus/isolation & purificationABSTRACT
The actions of histamine on pial venule leaky site formation were measured intravitally in two inbred strains of mice (BALB/c and SJL/J). Pial venules were visualized using a cranial window microscopy technique, and microvascular leaky site formation was assessed visually using a fluorescein-dextran indicator. SJL/J mice were found to be sensitive to histamine-induced leakage, whereas the BALB/c strain was refractory. Exposure to pertussis toxin enhanced the sensitivity to histamine in the SJL/J strain, but little effect was observed for BALB/c mice. However, the employment of a polymerase chain reaction (PCR) technique for the detection of mRNA for histamine H1 receptor identified receptor-specific message in isolated cerebrovascular endothelium from both strains of mice. The lack of pial responsiveness in the BALB/c mice remains unexplained. Mast cells in the dura mater were found to be more numerous in SJL/J mice than in BALB/c mice. This observation supports previous observations of strain-specific differences in CNS inflammation. The results support the concept that genetically controlled differences in vascular sensitivity and localization of CNS-associated mast cells may play important roles in the generation of vasogenic edema and inflammation in CNS trauma and disease.
Subject(s)
Blood-Brain Barrier/drug effects , Cerebral Veins/physiology , Histamine/pharmacology , Animals , Arterioles/anatomy & histology , Arterioles/drug effects , Base Sequence , Cerebral Veins/drug effects , Histamine H1 Antagonists/pharmacology , Mast Cells/drug effects , Mast Cells/metabolism , Mice , Mice, Inbred BALB C , Mice, Inbred Strains , Microscopy, Fluorescence , Molecular Sequence Data , Pertussis Toxin , RNA, Messenger/biosynthesis , Receptors, Histamine H1/biosynthesis , Species Specificity , Venules/anatomy & histology , Venules/drug effects , Virulence Factors, Bordetella/pharmacologyABSTRACT
This communication describes a relatively novel cell culture technique for the isolation of cerebrovascular endothelial cells from three strains of inbred mice. Cerebrovascular endothelial cells were identified by their morphology, the presence of Factor VIII-related antigen and angiotensin-converting enzyme, and the uptake of acetylated low-density lipoprotein. Cloned cerebrovascular endothelial cells were found to maintain their differentiated state and diploid genotype through 15 serial passages. The morphology and growth characteristics of these cells were found to be altered when cultured on different extracellular matrices. The isolation and cloning methods described are simple and highly reproducible.