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K Q Hu

Publications and source records attributed to K Q Hu.

At least 37 records · Page 2Linked to original sources
PubMed1991

Regulation of the hepatitis B virus gene expression by the enhancer element I.

Hepatitis B virus (HBV) enhancer is a cis-acting element capable of potentiating transcription with liver cell specificity. To assess the regulatory function of the enhancer during viral gene expression, we employed whole genome transfections in both liver- and non-liver-derived cell lines and compared them with those of enhancerless genome transfections. Using a transient scheme of expression, the effect of enhancer deletion on the transcription of HBV genes was investigated. S1 nuclease analysis showed that deletion of the enhancer sequences did not alter the specificity of the HBV transcripts. Also, the nuclear run-on analysis demonstrated that the influence of enhancer on the HBV gene expression is at the transcriptional level. We further noted that in the absence of the enhancer, synthesis of the pregenomic RNA which originates from the core/pregenomic promoter was markedly affected. This result then implicates an additional role of the enhancer during the replication of HBV.

Carcinoma, Hepatocellular↗
PubMed1991

Transcriptional factor C/EBP binds to and transactivates the enhancer element II of the hepatitis B virus.

The human hepatitis B Virus genome (HBV) contains a liver-specific enhancer upstream of the X ORF which has been studied in detail by several investigators. A second liver-specific enhancer element, designated here as enhancer II, has been relatively recently described in the HBV genome, which is located within the core/pregenomic promoter. We have studied the interactions of transcriptional factors with this element and show here that the nuclear factor CCAAT/enhancer binding protein (C/EBP) binds at a unique site within these sequences. Further, using the transient cotransfection scheme of expression with C/EBP encoding vectors and an enhancer II-reporter gene construct, we demonstrate that the enhancer element II responds to increasing amounts of C/EBP by displaying transactivation. Evidence for the functional role of the enhancer element II in transcriptional regulation of the HBV gene expression is presented. A major influence of the enhancer II appears to be on the surface antigen expression.

Base Sequence↗
PubMed1990

Modulation of the insulin growth factor II/mannose 6-phosphate receptor in microvascular endothelial cells by phorbol ester via protein kinase C.

Phosphorylation of hormone receptors by protein kinase C (PKC) may be involved in the regulation of receptor recycling. We have studied the recycling and the phosphorylation state of the insulin growth factor (IGF) II/mannose 6-phosphate (Man-6-P) receptor in microvascular endothelial cells from rat adipose tissue. Scatchard analysis showed these cells have over 2 x 10(6) receptors/cell with an affinity constant of 1 x 10(9) M-1. In the presence of phorbol myristate acetate (PMA), an activator of PKC and analog of diacylglycerol, IGF-II receptor number increased in the plasma membrane by 60% without changes in the binding affinity. This increase in cell surface receptor number was confirmed by affinity cross-linking and 125I-surface labeling studies, occurred with a half-time of 20 min, and was reversible upon withdrawal of PMA. The redistribution of IGF-II/Man-6-P receptors was not due to an inhibition of internalization which was in fact stimulated by PMA. The effect of PMA on IGF-II receptor recycling correlated with its stimulation of PKC activity. Furthermore, after down-regulation of cellular PKC levels by preincubation with PMA, PMA was unable to activate residual PKC activity in the membranous pool or increase IGF-II receptor number at the cell surface. The phosphorylation state of the IGF-II/Man-6-P receptor was determined by 32P labeling of intact cells and immunoprecipitation with anti-receptor antibodies. In the basal state, the receptor was phosphorylated only on serine residues which was increased by 75% after treatment with PMA. In contrast, IGF-II decreased receptor phosphorylation and plasma membrane binding in a parallel and dose-dependent manner. Thus, PKC-stimulated serine phosphorylation of IGF-II/Man-6-P receptor may promote the translocation of the receptor to the cell surface, whereas IGF-II-stimulated dephosphorylation of the receptor may lead to a decrease in the number of cell surface receptors. These data suggest a role for PKC-mediated serine phosphorylation in the regulation of intracellular trafficking of receptors in endothelial cells.

Adipose Tissue↗
PubMed1990

Multiple liver-specific factors bind to the hepatitis B virus core/pregenomic promoter: trans-activation and repression by CCAAT/enhancer binding protein.

The human hepatitis B virus (HBV) is a hepatotropic virus that replicates through an RNA intermediate referred to as the pregenome. The promoter that directs the synthesis of the pregenome and several other transcripts with heterogeneous 5' ends is of particular interest because of its role in regulating key functions during the viral life cycle. We have examined the liver-specific characteristics of this promoter by DNA-protein interactions and by demonstrating the in vivo function of the promoter using the luciferase reporter gene expression system. The DNA-protein interactions in this region appear to be almost entirely liver-specific. Among these, a liver-specific nuclear factor, CCAAT/enhancer binding protein, binds to at least five sites on this promoter. Transient cotransfection experiments using CCAAT/enhancer binding protein expression vectors and the core promoter in the context of either the native hepatitis B virus genome or the luciferase reporter gene demonstrate that CCAAT/enhancer binding protein at low concentration modestly activates expression from the core promoter but represses at high concentration.

Animals↗
PubMed1990

Trans-activation of HLA-DR gene by hepatitis B virus X gene product.

Hepatocellular injury during hepatitis B virus (HBV) infection has been postulated to result from a human leukocyte antigen (HLA)-restricted T-lymphocyte host immune response against HBV antigens. Although HLA expression is enhanced in the presence of hepatic inflammation, whether HBV itself can induce HLA expression on infected hepatocytes is unknown. In this study, we demonstrate the induction of HLA-DR expression on human hepatoma cell lines transfected with HBV DNA sequences. The HBV X gene alone was capable of inducing HLA-DR expression. This induction correlated with elevated HLA-DR RNA, and this resulted directly from transcriptional trans-activation of the HLA-DR gene by the HBV X protein. These studies suggest that the HBV X protein can regulate the expression of HLA-DR and thus raise the possibility of participation by the X gene in the immunopathogenesis of HBV infection.

Antigens, Viral↗
PubMed1990

A study of the binding ability of pre-S1 and S2 proteins of hepatitis B virus to human serum albumin.

To assess the relationship between pre-S proteins of HBV and polymerized human serum albumin (PHSA), a labeled avidin biotin ELISA was used to detect pre-S1, S2 and PHSA receptor (PHSAR) activity. PHSAR activity was only present in the samples positive for pre-S1 and/or S2, but not in the samples positive only for HBsAg. Pre-S1, S2 and PHSAR could in some degree be blocked by preincubating serum with PHSA, and the blocking efficiency of PHSA with respect to pre-S2 and PHSAR was similar, suggesting that pre-S2 is the dominant site for binding PHSA in vitro. We also found that PHSAR activity was detectable in 2 cases positive only for pre-S1, but not pre-S2. Furthermore, PHSA could selectively block pre-S1 and PHSAR activity in 2 cases negative for pre-S2, revealing that pre-S1 also possesses binding ability to PHSA, at least in a small number of cases. Using sandwich ELISA, we demonstrated the existence of complexes of HBV envelope proteins and human serum albumin (HSA) in some HBV infected serum samples. The possible significance of these complexes is discussed.

Enzyme-Linked Immunosorbent Assay↗
PubMed1989

Endothelin stimulates a sustained 1,2-diacylglycerol increase and protein kinase C activation in bovine aortic smooth muscle cells.

Endothelin is a long-lasting potent vasoconstrictor peptide. We report here that in bovine aortic smooth muscle cells, endothelin biphasically increased total cellular diacylglycerol (DAG) content. When cellular DAG was labeled with [14C] glycerol for 48h, endothelin stimulated [14C]DAG formation in a biphasic pattern. Only one prolonged phase of DAG accumulation was observed when cells were labeled with [3H]glycerol for 2 h. Endothelin induced an increase in the membranous protein kinase C (PKC) activities, which lasted for more than 20 min. These data suggest that (i) endothelin stimulates a sustained generation of DAG, (ii) this accumulation of DAG results in a sustained translocation of cytosolic PKC activities to the membrane.

Animals↗
PubMed1989

A preliminary study on expression and significance of pre-S1 protein in liver tissue of patients with HBV infection.

Rabbit antiserum to Pre-S1 protein was used to establish peroxidase-antiperoxidase (PAP) and avidin-biotin-peroxidase complex (ABC) immunohistochemical techniques for detection of Pre-S1 protein in paraffin-embedded liver tissue. Pre-S1 protein could be expressed in hepatocyte cytoplasm and on membrane in some cases with chronic viral hepatitis, cirrhosis and hepatocellular carcinoma (HCC), and its expression was intimately associated with HBsAg, HBcAg in liver and HBV DNA in serum, indicating that pre-S1 protein may represent the essential component of hepatitis B virus (HBV) and also serve as one of the markers of HBV infection. The incidence of Pre-S1 protein was slightly lower in nontumorous liver of HCC than in other cases and Pre-S1 protein could not be detected in tumorous tissue of HCC suggesting that expression of pre-S1 protein may be suppressed in HCC cases.

Hepatitis B↗
PubMed1989

Expression and clinical significance of pre-S1 and S2 proteins of HBV in sera of patients with chronic liver disease.

To assess the significance of pre-S proteins expression during chronic hepatitis B virus (HBV) infection, we developed a sensitive labeled avidin biotin ELISA to detect pre-S1 and pre-S2 proteins. In serum specimens from 80 patients with chronic liver disease, the frequency of pre-S1 and S2 proteins was 53.7% and 47.5%, respectively. Furthermore, it reached 87.8% and 77.6%, respectively, in cases with chronic HBV infection, indicating that pre-S proteins are usually expressed in sera with chronic HBV infection. We found that the expression of pre-S proteins is closely associated with HBV replicating markers, such as HBV DNA, HBcAg and HBeAg, in sera of patients with chronic HBV infection. Both pre-S1 and S2 proteins were often concurrently expressed in liver and serum with chronic HBV infection. However, the frequency was slightly higher in liver than in serum, suggesting that it may be clinically valuable to detect pre-S proteins in serum and liver simultaneously to determine the status of HBV infection. Our results also indicated that pre-S proteins expression in serum can serve as markers of HBV infection, but cannot be used to estimate the severity and activity of hepatic pathology.

Enzyme-Linked Immunosorbent Assay↗
PubMed1989

Characterization of endothelin receptors and effects of endothelin on diacylglycerol and protein kinase C in retinal capillary pericytes.

Retinal capillary pericyte is a cell type selectively lost in early diabetic retinopathy. The physiological function of pericytes is not yet clearly identified, although it probably has contractile properties. We determined the specific binding of endothelin 1, a 21-amino acid peptide with potent vasoconstrictive action, and the stimulation of diacylglycerol/protein kinase C (DAG/PKC) pathway in cultured retinal capillary pericytes by endothelin. A single specific binding site for 125I-labeled endothelin was identified, with an apparent Kd of 1.3 nM and a maximal binding capacity of approximately 1-2 x 10(5) sites/cell. Endothelin (100 nM) increased total cellular DAG content by 15% at 5 min and 24% at 10 min. When pericytes were labeled isotopically with [3H]glycerol, endothelin stimulated [3H]DAG formation by 100% at 10 min and 88% at 30 min. After 10 min of endothelin treatment, PKC activities were increased by 60 and 100% in the membranous and cytosolic pools, respectively. We conclude that bovine retinal capillary pericytes possess numerous high-affinity specific binding sites for endothelin that mediate the action of endothelin by the stimulation of the DAG/PKC pathway in pericytes. These findings suggest that endothelin is a regulator of the contractile properties of pericytes, which may be adversely affected in diabetic retinopathy.

Animals↗
PubMed1989

Intrahepatic expression of pre-S proteins of hepatitis B virus and its possible relation to liver cell necrosis.

To assess the significance of intrahepatic expression of pre-S1 and S2 proteins of hepatitis B virus (HBV) in patients with HBV infection, an indirect immunoperoxidase technique employed monoclonal antibodies to pre-S proteins was used to detect pre-S1 and -S2 proteins in 80 liver specimens. The frequency of pre-S1 and -S2 proteins was 61.3% and 51.3%, respectively, and the co-expression of pre-S and HBsAg occurred in most specimens. The preferential expression of pre-S1 and -S2 in HBcAg-positive specimens suggests that pre-S proteins are associated with HBV replication. Membranous expression of both pre-S1 and -S2 is associated with inflammatory activity and liver cell necrosis. Furthermore, our results show that T cells, not NK or B cells, were the predominantly infiltrating cells in necrotic foci with pre-S expression. Almost all of these T cells may express HLA-DR antigen simultaneously; therefore, they are activated. In conjunction with these data, we conclude that, as the essential components of HBV envelope proteins, pre-S proteins may play an important role in resulting in liver cell necrosis.

Adult↗
PubMed1989

[An immunohistochemical study on pre-S proteins of hepatitis B virus].

An indirect immunohistochemical technique was established to detect pre-S1 and pre-S2 proteins in liver with monoclonal antibodies, and altogether 80 samples of liver were studied. Under light microscope, pre-S1 and pre-S2 were known to be expressed in a similar way and might be divided into three patterns: diffusion type, inclusion type and membrane type. Membranous type expression of both pre-S1 and pre-S2 was associated with activity of the liver diseases and liver cell necrosis. Pre-S1 and Pre-S2 were also expressed in several samples of hepatocellular carcinoma.

Hepatitis B Surface Antigens↗