ABSTRACT: To investigate peripheral blood mononuclear cells (PBMCs) immune-related gene expression profile in patients with chronic virus hepatitis B(CHB) by oligonucleotide gene array technique.
PBMCs were collected from the members of a family of clustering hepatitis B virus (HBV) infection including 5 CHB patients and 4 healthy spouses and RNA prepared from PBMCs was hybridized to high-density oligonucleotide arrays(HG-U133A 2.0 Human Gene Chips, Affymetrix), covering the expression of 22 000 human ESTs. Primary scanned image was analyzed with DNT software package.
Out of the 22 000 ESTs, 24 different immune-related genes were identified. Among the 24 genes, 7 genes showed increased expression and 17 genes showed decreased expression in CHB compared with those in healthy spouses. The up-regulated genes were mainly associated with adaptive immunity, while the down-regulated genes were associated with innate immunity.
Our findings suggest that HBV infection alters a broad range of immunity genes expression and innate immunity-associated genes are important in the defense against HBV chronic infection.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 10/2006; 22(5):620-2.
ABSTRACT: To investigate the peripheral blood monocyte (PBMC) gene expression profile in a familial clustering of patients with chronic hepatitis B (CHB).
cRNA prepared from PBMC in a family with 5 CHB patients and 4 normal controls was hybridized to high-density oligouncleotide arrays (HG-U133A 2.0 Human GeneChips, Affymetrix), which interrogate the expression of approximately 22,000 human ESTs. Primary image obtained from scanning was analysed with a DNT software package. Real-time PCR was employed to confirm the gene chip results.
55 genes out of 22,000 ESTs were identified differently. Among the 55 genes 14 showed increased expression and 41 showed decreased expression in the familial clustering CHB patients compared with those in normal controls. Most of the genes (57%) were involved in immunity, inflammation, apoptosis, signaling transduction, and cell cycle.
These results suggest that the hosts with this broad range of gene expression alterations are susceptible to hepatic B infection.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 12/2005; 13(11):811-4.
ABSTRACT: To investigate the biological function of F protein by yeast two-hybrid system.
We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (alpha type) containing liver cDNA library plasmid in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection and screening. After extracting and sequencing plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.
Thirty-six colonies were selected and sequenced. Among them, 11 colonies were zymogen granule protein, 5 colonies were zinc finger protein, 4 colonies were zinc-alpha-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor I, 1 colony was vitronectin, and 2 colonies were new genes with unknown function.
The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins.
World Journal of Gastroenterology 10/2005; 11(36):5659-65. · 2.47 Impact Factor
ABSTRACT: To investigate the biological function of p7 protein and to look for proteins interacting with p7 protein in hepatocytes.
We constructed p7 protein bait plasmid by cloning the gene of p7 protein into pGBKT7, then transformed it into yeast AH109 (a type). The transformed yeast was mated with yeast Y187 (alpha type) containing liver cDNA library plasmid, pACT2 in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selection and screening. After extracting and sequencing of plasmids from blue colonies, we performed sequence analysis by bioinformatics.
Fifty colonies were selected and sequenced. Among them, one colony was Homo sapiens signal sequence receptor, seven colonies were Homo sapiens H19, seven colonies were immunoglobulin superfamily containing leucine-rich repeat, three colonies were spermatid peri-nuclear RNA binding proteins, two colonies were membrane-spanning 4-domains, 24 colonies were cancer-associated antigens, four colonies were nucleoporin 214 ku and two colonies were CLL-associated antigens.
The successful cloning of gene of protein interacting with p7 protein paves a way for the study of the physiological function of p7 protein and its associated protein.
World Journal of Gastroenterology 09/2005; 11(30):4709-14. · 2.47 Impact Factor