ABSTRACT
AIMS: We investigated the potential of apolipoprotein D (apoD) as cerebrospinal fluid (CSF) biomarker for cerebral amyloid angiopathy (CAA) after confirmation of its association with CAA pathology in human brain tissue. METHODS: The association of apoD with CAA pathology was analysed in human occipital lobe tissue of CAA (n = 9), Alzheimer's disease (AD) (n = 11) and healthy control cases (n = 11). ApoD levels were quantified in an age- and sex-matched CSF cohort of CAA patients (n = 31), AD patients (n = 27) and non-neurological controls (n = 67). The effects of confounding factors (age, sex, serum levels) on apoD levels were studied using CSF of non-neurological controls (age range 16-85 years), and paired CSF and serum samples. RESULTS: ApoD was strongly associated with amyloid deposits in vessels, but not with parenchymal plaques in human brain tissue. CSF apoD levels correlated with age and were higher in men than women in subjects >50 years. The apoD CSF/serum ratio correlated with the albumin ratio. When controlling for confounding factors, CSF apoD levels were significantly lower in CAA patients compared with controls and compared with AD patients (P = 0.0008). CONCLUSIONS: Our data show that apoD is specifically associated with CAA pathology and may be a CSF biomarker for CAA, but clinical application is complicated due to dependency on age, sex and blood-CSF barrier integrity. Well-controlled follow-up studies are required to determine whether apoD can be used as reliable biomarker for CAA.
Subject(s)
Apolipoproteins D/metabolism , Biomarkers/cerebrospinal fluid , Cerebral Amyloid Angiopathy/pathology , Aged , Cerebral Amyloid Angiopathy/metabolism , Female , Humans , Male , Middle AgedABSTRACT
Inactive nuclear factor kappaB (NF-kappaB) complexes are retained in the cytoplasm by binding to inhibitory proteins, such as IkappaBalpha. Various stimuli lead to phosphorylation and subsequent processing of IkappaBalpha in the 26S proteasome and import of the active NF-kappaB transcription factor into the nucleus. In agreement with our previous finding that p90(rsk1) is essential for TPA-induced activation of NF-kappaB in Adenovirus 5E1-transformed Baby Rat Kidney cells, we now report that the MEK/ERK/p90(rsk1) inhibitor U0126 efficiently blocks TPA-induced IkappaBalpha processing in these cells. However, in U2OS cells, the cytokine-inducible IkappaB kinase complex (IKK) is the essential component of the TPA signal transduction pathway. Activation of the IKK complex in response to TPA is mediated by PKC-alpha, since both the PKC inhibitor GF109203 and a catalytically inactive PKC-alpha mutant inhibit activation of endogenous IKK by TPA, but not by tumor necrosis factor-alpha (TNF-alpha). We conclude that IKK is an integrator of TNF-alpha and TPA signal transduction pathways in U2OS cells.
Subject(s)
I-kappa B Proteins , Isoenzymes/metabolism , NF-kappa B/metabolism , Protein Kinase C/metabolism , Protein Serine-Threonine Kinases/metabolism , Signal Transduction/drug effects , Tetradecanoylphorbol Acetate/pharmacology , Butadienes/pharmacology , DNA-Binding Proteins/metabolism , Enzyme Activation/drug effects , Genes, Dominant/genetics , Humans , I-kappa B Kinase , Indoles/pharmacology , Isoenzymes/antagonists & inhibitors , Isoenzymes/genetics , Kinetics , MAP Kinase Kinase 1 , Maleimides/pharmacology , Mitogen-Activated Protein Kinase Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinase Kinases/metabolism , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinases/metabolism , Multienzyme Complexes/drug effects , Mutation/genetics , NF-KappaB Inhibitor alpha , Nitriles/pharmacology , Protein Kinase C/antagonists & inhibitors , Protein Kinase C/genetics , Protein Kinase C-alpha , Protein Processing, Post-Translational/drug effects , Protein Serine-Threonine Kinases/antagonists & inhibitors , Protein Serine-Threonine Kinases/genetics , Ribosomal Protein S6 Kinases/antagonists & inhibitors , Ribosomal Protein S6 Kinases/metabolism , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Proteins encoded by non-oncogenic adenovirus type 5 and oncogenic adenovirus type 12 differentially affect expression of a number of cellular genes. We have used cDNA micro array analysis to identify a cellular gene that is expressed in Ad12- but not in Ad5-transformed cells. This cellular gene was found to be the gene encoding follistatin-related protein, a TGF-beta inducible gene. Consistently, a constitutive factor binding to Smad binding elements was found in adenovirus type 12-transformed cells.
Subject(s)
Adenoviruses, Human/genetics , Cell Transformation, Viral , Gene Expression Regulation , Glycoproteins/genetics , Cell Line, Transformed , Expressed Sequence Tags , Follistatin-Related Proteins , Gene Expression Regulation/drug effects , Humans , Kidney , Lung , Oligonucleotide Array Sequence Analysis , Retina , Species Specificity , Transforming Growth Factor beta/pharmacologyABSTRACT
The complex formation between glycoproteins H (gH) and L (gL) of herpes simplex virus type 1 (HSV-1) was studied by using five recombinant baculoviruses expressing open reading frames that contain deletions in the coding region of the extracellular domain of gH. In addition, the gH-deletion mutants contained a C-terminal tag. Complex formation of gL and the gH-deletion mutants was studied by immunoprecipitations with anti-tag monoclonal antibody (MAb) A16 and with the gH-specific MAbs 37S, 46S, and 52S. All gH-deletion mutants were complexed to gL when analyzed by MAb A16. MAb 37S precipitated complexes between gL and the two gH-deletion mutants that contain the epitope of this MAb. When the gH conformation-dependent MAbs 46S and 52S were used, gL was coprecipitated together with the gH-deletion mutant lacking amino acids 31-299, but gL was not coprecipitated with the gH-deletion mutant lacking amino acids 31-473. The data from the precipitation studies do allow at least two interpretations. There is either one site for gL binding on gH (residue 300-473) or gL contacts multiple regions of gH. We were unable to demonstrate gL-dependent cell surface expression of either of the gH-deletion mutants. This suggests that the coassociation of gH with gL is necessary but not sufficient for transport of gH to the cell surface.
