Document 0034
 DOCN  M9590034
 TI    Semi-quantitative analysis of cytokine gene expression in blood and
       cerebrospinal fluid cells by reverse transcriptase polymerase chain
       reaction.
 DT    9509
 AU    Rieckmann P; Albrecht M; Ehrenreich H; Weber T; Michel U; Department of
       Psychiatry, University of Gottingen, Germany.
 SO    Res Exp Med (Berl). 1995;195(1):17-29. Unique Identifier : AIDSLINE
       MED/95304169
 AB    An easy, reproducible and semi-quantitative, non-radioactive method for
       the analysis of mRNA expression for various cytokines, (i.e.,
       Interleukin (IL)-1 beta, IL-4, IL-6, tumor necrosis factor (TNF)-alpha,
       lymphotoxin (LT), transforming growth factor (TGF)-beta, interferon
       (IFN)-gamma and endothelin-1 (ET-1)) in cells from cerebrospinal fluid
       (CSF) and peripheral blood mononuclear cells (PBMC) has been
       established. By means of polymerase chain reaction primers that cover a
       splice junction, amplification of contaminating DNA was omitted.
       Densitometric scanning of ethidium bromide-stained agarose gels proved
       to be very sensitive for semiquantitative analysis of PCR products.
       Serial tenfold dilutions of cDNA revealed a log-linear regression from
       10(6) to 10(2) cells under optimal cycle conditions. The intra- and
       inter-assay variability of the method was below 10%. With this assay,
       the cytokine expression pattern of as few as 10(4) mononuclear cells
       from blood or CSF was determined. This method made it possible to detect
       differences in the cytokine gene expression pattern of mononuclear cells
       from patients with different neurological diseases. CSF cells from 43
       patients with various neurological diseases were analyzed. TNF-alpha,
       LT, and IL-1 mRNA were prominent in the CSF cells of most patients with
       bacterial meningitis. TNF-alpha, LT, IFN-gamma and IL-6 mRNAs were
       detected in patients with active multiple sclerosis, whereas TNF-alpha,
       IL-6, and endothelin-1 mRNA expression was found frequently in patients
       with HIV encephalitis. Pro-inflammatory cytokines were rarely detected
       in CSF cells from patients with non-inflammatory diseases of the central
       nervous system. In blood mononuclear cells from patients with multiple
       sclerosis, TNF-alpha mRNA expression was associated with disease
       activity. The sensitivity, specificity, velocity and reliability of this
       assay considerably facilitates the analysis of cytokine production in
       mononuclear cells even in conditions where only a limited number of
       cells is available for analysis.
 DE    Base Sequence  Cerebrospinal Fluid/CHEMISTRY
       Cytokines/*BLOOD/*CEREBROSPINAL FLUID/GENETICS  DNA,
       Complementary/STANDARDS  Gene Expression/*IMMUNOLOGY  Human  Molecular
       Sequence Data  Polymerase Chain Reaction/*METHODS  Predictive Value of
       Tests  RNA, Messenger/ANALYSIS  Support, Non-U.S. Gov't  Tumor Necrosis
       Factor/ANALYSIS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).