Document Detail


Isolation of brain parenchymal lymphocytes for flow cytometric analysis. Application to acute viral encephalitis.
MedLine Citation:
PMID:  1675228     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A strategy for the isolation of mononuclear cells from the brain parenchyma of mice with ongoing central nervous system (CNS) inflammation has been developed in order to permit flow cytometric (FCM) analysis of these cell populations. Sindbis virus (SV) encephalitis in mice is characterized morphologically by an infiltration of mononuclear cells into both brain parenchyma and cerebrospinal fluid (CSF). Perfused brain tissue from infected animals is collected, homogenized, and subjected to a mild enzymatic digestion. A sedimentation at unit gravity is performed to remove any large particulate debris, and the remaining tissue is then centrifuged over a modified density gradient which separates intact cells from smaller tissue fragments. Cells collected directly from these gradients can be stained with monoclonal antibodies and analyzed by FCM without further manipulation. Data generated by this method correlates with previous studies of SV encephalitis using immunohistochemical analysis of brain tissue sections to quantify mononuclear cell types. This suggests that representative samples of the cellular infiltrate are obtained using this technique. The approach however, offers the possibility of more sophisticated and quantitative analyses of CNS inflammatory cells which is unobtainable by tissue section staining.
Authors:
D N Irani; D E Griffin
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of immunological methods     Volume:  139     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1991 Jun 
Date Detail:
Created Date:  1991-07-16     Completed Date:  1991-07-16     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  223-31     Citation Subset:  IM    
Affiliation:
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal
Antigens, CD3
Antigens, Differentiation, T-Lymphocyte / analysis
Antigens, Ly / analysis
Brain / immunology,  pathology*
CD4-Positive T-Lymphocytes / immunology
Cell Separation / methods
Centrifugation, Density Gradient
Encephalitis, Arbovirus / immunology*,  pathology
Flow Cytometry
Lymphocytes / cytology*
Mice
Myelin Sheath / ultrastructure
Receptors, Antigen, T-Cell / analysis
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, CD3; 0/Antigens, Differentiation, T-Lymphocyte; 0/Antigens, Ly; 0/Receptors, Antigen, T-Cell

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