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Biomedical subjects

M Yu

Publications and source records attributed to M Yu.

At least 37 records · Page 2Linked to original sources
PubMed2005

Enhanced maturation and functional capacity of dendritic cells induced by mannosylated L2 domain of ErbB2 receptor.

The nature of antigens and functional state of dendritic cells (DC) are important in antigen presentation. The ability of DC for the induction of T-cell responses is promoted by maturation. It has been confirmed that mannose receptors mediate highly efficient endocytosis and presentation of mannosylated proteins. In the present study, L2 domain of ErbB2 ectodomain was expressed in Escherichia coli, purified and mannosylated. The maturation and functional capacity of DC induced by mannosylated L2 (mL2) protein were investigated. The results showed that L2 protein could induce DC maturation, which was accompanied by elevated expression of MHC and co-stimulatory molecules. The effect of mL2 protein on DC maturation was more remarkable than that of non-mL2 proteins. Uptake of mL2 antigens by DC was more efficient. Furthermore, the T cells can be stimulated to proliferate in vitro and secrete Th1 and Th2 cytokines. Higher levels of both IFN-gamma and IL-10 were detected from the T cells stimulated by mL2-pulsed DC, suggesting a concurrent activation of CD4+ and CD8+ T cells. The results demonstrated that L2 domain of ErbB2 receptor is an immunodominant molecule. The mL2 domain of ErbB2 can induce an enhanced maturation and functional capacity of DC. It may become an effective strategy to induce anti-ErbB2 response.

Antigen Presentation↗
PubMed2005

Partial molecular characterization, polymorphism and chromosomal localization of the porcine PSMD4 gene.

The complete coding sequence of porcine PSMD4 gene encoding proteasome 26S non-ATPase subunit 4 was obtained by the reverse transcriptase-polymerase chain reaction (RT-PCR) and deposited in GenBank nucleotide database (AY704418). The nucleotide sequence and the predicted protein sequence shared the high sequence identity with their mammalian counterparts. A G/A single nucleotide polymorphism in intron 9 detected as Hin6I PCR-restriction fragment length polymorphism (PCR-RFLP) shows allele frequency differences between Meishan, Erhualian, Tibetan, Large White and Duroc breeds. Analyses of somatic cell hybrid panel (SCHP) and radiation hybrid (IMpRH) panel revealed that PSMD4 gene maps to SSC 4q21-q23 and closely linked the SW512 (14 cR, LOD = 19.47).

Amino Acid Sequence↗
PubMed2004

Therapeutic effects of dengue 2 virus capsid protein and staphylococcal nuclease fusion protein on dengue-infected cell cultures.

Dengue infection poses a serious public health problem in most tropical and subtropical areas. No effective antiviral drugs or vaccines are currently available against dengue infection. To explore the feasibility of using capsid-targeted viral inactivation (CTVI) as an antiviral strategy against dengue infection, we constructed a plasmid expressing a fusion protein consisting of staphylococcal nuclease (SN) fused to dengue 2 virus capsid protein (D2C), and investigated its effects on the production of infectious virions when introduced into BHK cells infected with dengue virus. The results indicated that D2C-SN can be expressed and tolerated in this mammalian cell culture. The enzymatically active SN moiety was incorporated into nascent virions during the process of viral assembly. By comparing the effects of incorporated SN and SN*, an enzymatically inactive missense mutant form of wild-type SN, on the infectivity of progeny virions, we clearly demonstrated that nucleolytic activity was the major antiviral mechanism. Expression of D2C-SN fusion protein as a therapeutic agent resulted in a reduction in infectious titers of 12- to 60-fold. Therefore, dengue virus may be particularly vulnerable to a CTVI therapeutic approach.

Animals↗
PubMed2004

Structural analysis of the epitopes on erbB2 interacted with inhibitory or non-inhibitory monoclonal antibodies.

erbB2 oncogene encodes a growth factor receptor. The overexpression of erbB2 was correlated with more aggressive tumors and a poorer prognosis. Some antibodies directed to this molecule have an antitumor effect in vivo, but some antibodies do not. In an attempt to understand the molecular basis of the anti-erbB2 antibody interaction with erbB2 ectodomain (ECD), we analyzed binding epitopes on erbB2 for inhibitory and non-inhibitory antibodies, Herceptin and HF by computer-guided protein engineering and site-directed mutagenesis. Two different interaction domains were identified by molecular docking, computer graphics and distance geometry method and confirmed through studies on a series of mutants of erbB2 ECD. Non-inhibitory antibody HF only recognized N-terminal portion of erbB2 ECD, but inhibitory antibody Herceptin bound to C-terminal portion of it exclusively. The region interacted with inhibitory antibody Herceptin can be an important target for anticancer therapies.

Antibodies, Monoclonal↗
PubMed2004

Retroviral delivery of GAD-IgG fusion construct induces tolerance and modulates diabetes: a role for CD4+ regulatory T cells and TGF-beta?

Previous studies have demonstrated that antigen-specific tolerance could be induced by lipopolysaccharide (LPS)-stimulated B cells retrovirally transduced with an immunoglobulin-antigen (or epitope-containing peptide) fusion construct. To investigate the mechanism of this gene therapy system, we now adapted this approach to immunotherapy of spontaneous diabetes in nonobese diabetic (NOD) mice, a T-cell-mediated autoimmune disease triggered, in part, by a pathogenic response to glutamate decarboxylase (GAD) 65. We demonstrate that LPS-stimulated splenocytes, retrovirally transfected with GAD-IgG fusion construct, induce a significant antigen-specific hyporesponsiveness at both cellular and humoral levels and reduce the incidence of diabetes in female NOD mice. Parallel with disease protection, we observed a prolonged increase of the numbers of CD4+CD25+ T cells in the periphery of GAD-IgG-treated mice, compared to those treated with a control IgG vector, both in the prediabetic period and persisting even 8 months after gene therapy. This increase appeared to be induced by the repeated stimulation of the antigen in the periphery instead of a result of differentiation of T-cell precursor in the thymus. Moreover, CD4+CD25+ T cells induced by GAD-IgG fusion construct were capable of suppressing the proliferative response of CD4+CD25- T cells in vitro; and ablation of the activity of CD4+CD25+ T cells by blocking antibody against CD25 could reverse GAD-specific T-cell hyporesponsiveness. These results suggested that CD4+CD25+ T-cell subset induced in GAD-IgG-treated NOD mice represented the regulatory or suppressive CD4+CD25+ T cells (Treg) and might play an important role in the induction and maintenance of tolerance in NOD mice. Furthermore, the numbers of splenic CD4+CD62L+ regulatory T cells in GAD-IgG-treated mice during the prediabetic period and serum TGF-beta levels in 34-38-week-old GAD-IgG-protected mice were also increased, compared to control IgG-treated ones. Therefore, we propose that the induction of tolerance and the prevention of diabetes incidence in NOD female mice induced by the GAD-IgG fusion construct may require CD4+ regulatory T cells, and the possible mediation of TGF-beta.

Animals↗
PubMed2004

Dynamic analysis of hepatitis B virus DNA and its antigens in 2.2.15 cells.

The 2.2.15 cells-derived from HepG2 cells transfected with a plasmid containing hepatitis B virus (HBV) DNA secrete surface antigen (HBsAg) particles, nucleocapsids and virions (Proc Natl Acad Sci U S A 1987; 84: 1005-1009). The latter elicit acute hepatitis in chimpanzees (Proc Natl Acad Sci U S A 1987; 84: 4641-4644). We studied the presence of intracellular and extracellular HBV covalently closed circular (ccc) DNA in this culture system by polymerase chain reaction (PCR), kinetically analysed HBsAg and hepatitis B e antigen (HBeAg) released in the culture media by quantitative enzyme-linked immunosorbent assay and quantitated by real-time PCR but HBV DNA from intracellular and extracellular HBV-DNA. HBV cccDNA was found both intracellularly and extracellularly. A significant correlation was seen between the extracellular HBV DNA levels and virus antigens (r = 0.833; P = 0.01 and r = 0.939; P < 0.01 for HBsAg and HBeAg, respectively), whereas there was no statistical correlation between intracellular HBV DNA levels and virus antigen levels (r = 0.024; P = 0.955 and r = 0.177; P = 0.625 for HBsAg and HBeAg, respectively). These data would be valuable in studies of the HBV life cycle and of potential anti-viral agents.

Cell Line, Tumor↗
PubMed2004

Sequence characterization, polymorphism and chromosomal localizations of the porcine PSME1 and PSME2 genes.

The full-length cDNA of porcine genes (PSME1 and PSME2) encoding proteasome activators PA28alpha- and beta-subunits were obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with their mammalian counterparts. The reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that porcine PSME1 and PSME2 genes are expressed in all eight tissues studied (liver, spleen, bladder, small intestine, kidney, heart, skeletal muscle and lung). The full-length genomic DNA of the porcine PSME1 and PSME2 genes were amplified by PCR. These two genes shared the same structure and were similar in size. A C/T single nucleotide polymorphism in PSME1 intron 8 detected as an SphI PCR-restriction fragment length polymorphism (PCR-RFLP) shows allele frequency differences between Meishan, Tibetan, Large White, Qingping, and Duroc pigs. The association analysis using two experimental GY selection lines selected for growth rate or leanness suggested that the PSME1 genotype was associated with weaning weight. Analyses of somatic cell hybrid (SCHP) and radiation hybrid (IMpRH) panels revealed that both genes map to SSC7q15.3-q21 and closely linked to the T-cell receptor alpha (TCRA) gene.

Animals↗
PubMed2004

Genetic diversity analyses of 10 indigenous Chinese pig populations based on 20 microsatellites.

To study the genetic diversity of Chinese indigenous pig breeds, a total of 403 pigs from 10 local populations and 1 exotic Duroc breed were genotyped for 20 microsatellite markers. Heterozygosity and Wright's F-statistics (F(IS), F(ST), and F(IT)) were calculated to determine the genetic variation in those populations. The observed heterozygosities were in the range of 0.31 (Duroc) to 0.66 (Shengxian). The F(IS) value was in a range of -0.07 to 0.48. The mean F(ST) showed that approximately 78% of the genetic variation was within-population and 22% was across the populations. The 10 Chinese local breeds were classified into two major groups according to the phylogenetic tree, which was based on standard genetic distance. Four pig populations, Jianli, Ganxi Two Ends Black, Shaziling, and Dongshan were grouped into one branch. Before the study, these four populations were all classified as Central China Two Ends Black according to coat color, shape of the head, and shape of the ear. The Jinhua pig, which also has the two-ends-black coat color, was also grouped to the same branch but was not traditionally classified into this type. The five populations were located in various provinces in central China. The other five populations, Nanyang Black, Hainan Spotted, Huainan Black, Jiaxing Black, and Shengxian Spotted (black body, white feet), were grouped into another branch. The two groups of pig breeds had the same F(ST) value (0.14) when calculated separately. This value was similar to that of Iberian pigs (0.13) but smaller than that of the European pigs (0.27) as reported by other researchers. Our study showed that large genetic differentiation exists in Chinese pig breeds. The grouping of the five two-ends-black populations into one branch of the phylogenetic tree may indicate that the number of conservation farms can be decreased for this type of pig.

Animals↗