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W Zou

Publications and source records attributed to W Zou.

At least 55 records · Page 3Linked to original sources
PubMed1999

Conformational epitope of the type III group B Streptococcus capsular polysaccharide.

The protective epitope of the type III group B streptococcal polysaccharide (GBSPIII) is length dependent and conformational. To obtain a more accurate characterization of the conformational epitope, ELISA inhibition and surface plasmon resonance studies were conducted on two GBSPIII-specific mAbs using a large panel of oligosaccharide probes. The results of the studies confirmed that 2 repeating units (RU) is the minimum binding unit and that, while increases in chain length from 2 RU to 7 RU caused further optimization of the epitope, it remained monovalent. A 3-fold increase in affinity was observed between 7 RU and 20 RU, which, by surface plasmon resonance studies on a Fab, was shown to be due to both further optimization of the individual epitope and the occurrence of multivalency of epitope. The data support our hypothesis that the conformational epitope is an extended helical segment of the GBSPIII. GBSPIII exists mainly in the random coil form, which structurally mimics short oligosaccharide self Ags, but it can infrequently and spontaneously form extended helices. Although not prevalent in GBSPIII, the immune system preferentially selects these helical epitopes because they are unique to the polysaccharide. Contrary to a previously proposed model of GBSPIII binding in which the binding of the first Ab propagates a continuum of helical epitopes, our binding kinetics are consistent only with the helical epitope's being discontinuous and infrequent.

Animals↗
PubMed1999

Antigen receptor engagement selectively induces macrophage inflammatory protein-1 alpha (MIP-1 alpha) and MIP-1 beta chemokine production in human B cells.

We show herein that B cell Ag receptor (BCR) triggering, but not stimulation by CD40 mAb and/or IL-4, rapidly induced the coordinated expression of two closely related T cell chemoattractants, macrophage inflammatory protein-1 beta (MIP-1 beta) and MIP-1 alpha, by human B cells. Naive, memory, and germinal center B cells all produced MIP-1 alpha/beta in response to BCR triggering. In contrast to MIP-1 alpha/beta, IL-8, which is spontaneously produced by germinal center B cells but not by naive and memory B cells, was not regulated by BCR triggering. Culturing follicular dendritic cell-like HK cells with activated B cells did not regulate MIP-1 alpha/beta production, but it did induce production of IL-8 by HK cells. Microchemotaxis assays showed that CD4+CD45RO+ T cells of the effector/helper phenotype actively migrated along a chemotactic gradient formed by BCR-stimulated B cells. This effect was partially blocked by anti-MIP-1 beta and anti-CC chemokine receptor 5 Ab, but not by anti-MIP-1 alpha Ab suggesting that MIP-1 beta plays a major role in this chemoattraction. Since maturation of the B cell response to a peptide Ag is mostly dependent on the availability of T cell help, the ability of Ag-stimulated B cells to recruit T cells via MIP-1 alpha/beta, may represent one possible mechanism enabling cognate interactions between rare in vivo Ag-specific T and B cells.

Antibodies, Anti-Idiotypic↗
PubMed1999

Acute upregulation of CCR-5 expression by CD4+ T lymphocytes in HIV-infected patients treated with interleukin-2. ANRS 048 IL-2 Study Group.

BACKGROUND: The treatment of HIV-infected patients with interleukin (IL)-2 causes a sustained increase in CD4+ T-lymphocyte counts, involving both naive and memory cells. However, the short-term immunological effects of IL-2, which may shed light on the mechanism of immune reconstitution by this cytokine, are unknown. OBJECTIVE: To evaluate the acute effect of IL-2 on circulating T-lymphocyte subpopulations and their expression of chemokine receptors. DESIGN AND METHODS: Flow cytometry, reverse transcriptase polymerase chain reaction and chemokine receptor function experiments were performed before and after 5 days of IL-2 administration in 30 HIV-infected patients. RESULTS: IL-2 induced an acute lymphopenia of both naive and memory T-helper (TH) lymphocytes. This was associated with a large increase in CC-chemokine receptor (CCR)-5 and CCR-2b expression by TH cells. Before IL-2 treatment, CCR-5 was mostly produced by CD62L- memory TH lymphocytes. After 5 days of IL-2 administration, the level of CCR-5 mRNA in circulating cells was 18.6 times higher than before treatment (P < 0.002). CCR-5 expression was upregulated in CD62L- memory TH lymphocytes, but also in CD62L+ memory and in naive (CD62L+ CD45RO-) TH lymphocytes. IL-2 treatment also increased the function of CCR-5 in TH cells. CONCLUSIONS: Chemokine receptors are involved in trafficking of lymphocytes. The IL-2-induced upregulation of chemokine receptors in TH cells may thus play a role in the acute effects of this cytokine in TH lymphocyte redistribution.

Adult↗
PubMed1999

Synthesis and NMR assignments of galactosylgloboside and its beta-D-GalNAc-(1-->4)-alpha-D-Gal-linked positional isomer in a conjugatable form.

Two pentasaccharides suitable for conjugation, namely 3-aminopropyl glactosylgloboside and its beta-D-GalNAc-(1-->4)-alpha-D-Gal-linked positional isomer, were synthesized from 3III,4III-di-O-unprotected globotrioside and the trichloroacetimidate of beta-D-Gal-(1-->3)-beta-D-GalNPhth derivative. Glycosylation at both positions led to the formation of beta-D-GalNPhth-(1-->4)-alpha-D-Gal and beta-D-GalNPhth-(1-->3)-alpha-D-Gal-linked products in a ratio of 1:1 without selectivity. Complete NMR spectral assignments are also described.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1999

Allylmalonamide as a bivalent linker: synthesis of biantennary GM3-saccharide--keyhole limpet hemocyanin glycoconjugate and the immune response in mice.

A biantennary GM3-saccharide (sialyllactoside) derivative (4) was constructed using allylmalonic acid as a bivalent linker, both carboxylic acids of which were condensed with 3-aminopropyl lactoside (2) prior to enzymatic sialylation with a fusion enzyme. While ozonolysis of its allyl group generated a saccharide having a terminal aldehyde (6), we were unable to couple 6 directly to protein by reductive amination. However, extension of the spacer by means of introducing a maleimide group to 6 through its aldehyde group to give 7 enabled the latter to be successfully coupled to thiolated proteins. The average ratios of saccharide to protein were observed to be 35 in KLH conjugate (13) and 9-12 in HSA conjugates (14 and 15). The antisera obtained by immunizing mice with the biantennary sialyllactoside-KLH conjugate (13) together with MPL adjuvant were analyzed by ELISA. Using several structurally related saccharide-HSA conjugates as screening antigens, it was concluded that anti-sialyllactoside antibodies, both IgG and IgM, were effectively raised. This was further supported by competitive inhibition experiments using lactoside (1), sialyllactoside (8) and biantennary sialyllactoside (4) as inhibitors.

Adjuvants, Immunologic↗
PubMed1999

Determination of interaction mechanism of sensorgrams by analysis of binding kinetics.

We monitored the interactions between pairs of molecules where the antigen adenylate kinase (AK) was immobilized on the surface of a chip and the antibody against AK, McAb3D3, was in solution. The association data that we obtained were not always accurately described by the expected pseudo-first-order reaction mechanism. A better description of the association data was achieved with a double-exponential function. Various models were applied to describe these observations: mass transport-controlled processes, inhomogeneous immobilized ligands, or inhomogeneous soluble analytes. Inhomogeneous immobilized ligands seemed to be the most likely explanation for the observed biphasic association kinetics. We simulated the kinetics of the SPR signal under the above-mentioned conditions. Plots of dR/dt versus R of the association phase showed characteristic differences between those nonlinearities resulting from mass transport limitation and those from inhomogeneous ligands. The plots of dR/dt versus R of McAb3D3 binding to immobilized AK show positive curvatures, indicating that the observed biphasic association kinetics is due to the inhomogeneity of the immobilized ligands. This is consistent with the results obtained from the comparison of various model fittings.

Adenylate Kinase↗
PubMed1999

The significance of anti-endothelial cell antibodies in patients with lupus nephritis and immunoblotting analysis of the target components.

OBJECTIVE: To investigate the prevalence of anti-endothelial cell antibodies and its possible role in the pathogenesis in lupus nephritis. METHODS: Sera from 58 patients with lupus nephritis were studied. Cellular enzyme linked immunoabsaborbent assay was used to detect anti-endothelial cell antibodies, and immunoblotting technique was performed to determine its specific endothelial target antigens. RESULTS: 36.2% of the patients with lupus nephritis had elevated IgG isotype anti-endothelial cell antibodies. The association was revealed between the presence of anti-endothelial cell antibodies (AECA) and some laboratory and renal biopsy features, such as thrombocytopenia, higher serum IgG level, lower serum C3 level, higher erythrocyte sedimentation rate (ESR), presence of anti-nuclear antibody (ANA), anti-dsDNA antibodies and glomerular thrombosis. Anti-endothelial cell antibodies were consisted of a group of heterogeneous antibodies, and reacted with different endothelial cell antigens which ranged from 12-90 kD in molecular size in immunoblot. A correlation was demonstrated between heavy proteinuria (> 3.5 g/24 hr) and the existence of antibodies against both 27 and 29 kD endothelial cell antigens, an association between the presence of glomerular capillary thrombi and antibody against the 29 kD endothelial cell antigen was also found. CONCLUSIONS: Lupus nephritis patients with anti-endothelial cell antibodies showed a complex autoimmune disorder and clinical disease activity, and the results also provided an indirect evidence that anti-endothelial cell antibodies might contribute to the pathogenesis of lupus nephritis.

Autoantibodies↗
PubMed1999

[Expression of endothelin 1 and tumor necrosis factor alpha in injuried renal tubules and their influences on renal interstitial fibroblasts].

OBJECTIVE: To study the expression of endothelin 1 (ET-1) and tumor necrosis factor (TNFalpha) in the epithelial cells of normal and injured renal tubules and their influences on renal interstitial fibroblasts. METHODS: Cultivation of renal tubular epithelial cells and renal interstitial fibroblasts and establishment of a renal tubulo-interstitial fibrosis (TIF) animal model; reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical staining, radioimmunoassay (RIA), double immunohistochemical; and (3)H-TDR incorporation techniques were applied to study the relationship between ET-1, plus TNFalpha and the proliferation of renal interstitial fibroblasts. RESULTS: Expression of ET-1 mRNA and TNFalpha mRNA from renal tubular epithelial cells was obtained (546 bp and 415 bp) respectively as well as the relevant ET-1 and TNFalpha proteins. The concentration of ET-1 and TNFalpha in the culture medium were 1.42 pg/ml and 0.58 ng/ml respectively. The amount of ET-1 and TNFalpha was increased during cell injury and regeneration. In addition, the ratio of (3)H-TDR incorporation and the expression of ET-R mRNA and TNF-R1 mRNA (545 bp and 347 bp) were markedly increased than that of the control group (P < 0.05) when ET-1 or TNFalpha was added in the culture media. CONCLUSION: Injured and regenerated renal tubular epithelial cells are able to synthesize and liberate more ET-1 and TNFalpha than that of the normal renal tubules. ET-1 and TNFalpha are also effective in promoting the proliferation of renal interstitial fibroblasts through ET-R and TNF-R.

Animals↗
PubMed1998

Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Sträussler-Scheinker disease.

The clinicopathological phenotype of the Gerstmann-Sträussler-Scheinker disease (GSS) variant linked to the codon 102 mutation in the prion protein (PrP) gene (GSS P102L) shows a high heterogeneity. This variability also is observed in subjects with the same prion protein gene PRNP haplotype and is independent from the duration of the disease. Immunoblot analysis of brain homogenates from GSS P102L patients showed two major protease-resistant PrP fragments (PrP-res) with molecular masses of approximately 21 and 8 kDa, respectively. The 21-kDa fragment, similar to the PrP-res type 1 described in Creutzfeldt-Jakob disease, was found in five of the seven subjects and correlated with the presence of spongiform degeneration and "synaptic" pattern of PrP deposition whereas the 8-kDa fragment, similar to those described in other variants of GSS, was found in all subjects in brain regions showing PrP-positive multicentric amyloid deposits. These data further indicate that the neuropathology of prion diseases largely depends on the type of PrP-res fragment that forms in vivo. Because the formation of PrP-res fragments of 7-8 kDa with ragged N and C termini is not a feature of Creutzfeldt-Jakob disease or fatal familial insomnia but appears to be shared by most GSS subtypes, it may represent a molecular marker for this disorder.

Adult↗
PubMed1998

Administration of interleukin 13 to simian immunodeficiency virus-infected macaques: induction of intestinal epithelial atrophy.

Increase Th2 cytokine production may contribute to some clinical manifestations of HIV infection, and studies have suggested that IL-13 rather than IL-4 is involved in these conditions. We directly tested this hypothesis by administrating IL-13 to SIV-infected macaques. SIV-infected rhesus macaques received a daily subcutaneous injection for 21 days of either IL-13 (10 microg/kg/day) or a placebo. The four macaques treated with IL-13 experienced body weight loss (9.95 +/- 0.71%) related to intestinal tract damage: they all suffered from a complete atrophy of duodenal villi. This was presumably due to premature epithelial cell death: proliferating Ki67+ cells in glandular crypts were as numerous as in control animals, but many epithelial cells developed apoptosis. The duodenal mucosa was infiltrated with cells expressing CD56 and PEN5, two markers of NK cells, and there was a deregulation of local cytokine and chemokine production characterized by a decrease in IL-10 gene expression (25% of controls) and an increase in gene expression for IFN-gamma (4-fold control), MIP-1alpha (8-fold control), and MIP-1beta (13-fold control). Thus, IL-13 can induce digestive epithelial cell injury in vivo in primates infected with a retrovirus. Therefore, its role should be considered in digestive manifestations of HIV infection as well as in other disorders associated with intestinal epithelial atrophy.

Animals↗
PubMed1998

Oligosaccharide fragments of the type III group B streptococcal polysaccharide derived from S. pneumoniae type 14 capsular polysaccharide by a chemoenzymatic method.

Partial N-deacetylation fo the GlcNAc residues in S. pneumoniae type 14 capsular polysaccharide (Pn14-PS) backbone was achieved by treatment with base, and the product was subsequently enzymatically sialylated at the 3-O-positions of the terminal galactose residues. The resultant, partially N-deacetylated type III Group B streptococcus capsular polysaccharide (GBSIII-PS) was subjected to nitrous acid deamination, which resulted in the degradation of GBSIII-PS polysaccharide into oligosaccharides containing increasing numbers of the identical repeating units. The oligosaccharides were then separated by passage through a Superdex 30 column and characterized by ESIMS and NMR spectroscopic analysis.

Bacterial Capsules↗
PubMed1998

[The evaluation of electron microscopy in the pathological diagnosis of renal biopsies].

OBJECTIVE: To evaluate the use of electron microscopy(EM) in pathological diagnosis of renal biopsies. METHOD: 777 cases of renal biopsy in Department of Nephrology, First Hospital of Beijing Medical University from Jan 1995 to Jun 1997 were studied. We compared the preliminary diagnosis by light microscopy(LM) and immuno-fluorescence(IF) with the final diagnosis by electron microscopy(EM). The use of EM in the diagnosis was divided into three parts. EM was needed to make a correct final diagnosis, provided confirmatory data or additional information relevant to the diagnosis and was complementary to LM and IF. RESULTS: Among 777 cases of renal biopsy, EM was needed to make a correct final diagnosis in 18.5%, and provided important referential information in 13.5%. CONCLUSION: EM provided useful diagnostic information in about one third cases of renal biopsy.

Biopsy, Needle↗
PubMed1997

Production and roles of IL-6, IL-10, and IL-13 in B-lymphocyte malignancies and in B-lymphocyte hyperactivity of HIV infection and autoimmunity.

We analyzed the production and the roles of IL-6, IL-10, and IL-13 in B-lymphoid malignancies and in specific diseases with B-lymphocyte hyperactivity. Both IL-13 and IL-10 genes are expressed in B-cell lymphomas. However, their contribution to tumor progression is unclear. In certain lymphoproliferative disorders that develop in transplanted patients, IL-6 is produced by malignant cells and is a major factor of their proliferation. In other lymphomas, the IL-6 gene is expressed only in malignancies where differentiated malignant cells are present. In these lymphomas, IL-6 is produced by stromal cells, and the malignant cells express the IL-6 receptor. In patients with HIV infection, the level of production of IL-6, IL-10, and IL-13 is not higher than those of other conditions with immune activation. However, IL-6 contributes to increased production of IgG and IgA in vivo. In Castleman's disease, IL-6 is produced in the lymph node germinal centers, partly originating from follicular dendritic cells, which may explain some of the pathogenesis of this disease. In systemic lupus erythematosus, the critical cytokine is IL-10, which is produced in large amounts by B lymphocytes and monocytes and is responsible for autoantibody production. Taken together, these data emphasize the roles of IL-6 and IL-10, usually produced by nonlymphoid cells, on B lymphocytes, either malignant or hyperactivated.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Allelic origin of the abnormal prion protein isoform in familial prion diseases.

The hallmark of prion diseases is the presence of an aberrant isoform of the prion protein (PrP(res)) that is insoluble in nondenaturing detergents and resistant to proteases. We investigated the allelic origin of PrP(res) in brains of subjects heterozygous for the D178N mutation linked to fatal familial insomnia (FFI) and a subtype of Creutzfeldt-Jakob disease (CJD178), as well as for insertional mutations associated with another CJD subtype. We found that in FFI and CJD178 subjects, only mutant PrP was detergent-insoluble and protease-resistant. Therefore, PrP(res) derives exclusively from the mutant allele carrying the D178N mutation. In contrast, in the CJD subtype harboring insertional mutations, wild-type PrP was also detergent-insoluble and likely to be protease-resistant. Our findings indicate that the participation of the wild-type PrP in the formation of PrP(res) depends on the type of mutations, providing an insight into the molecular mechanisms underlying the phenotypic heterogeneity in familial prion diseases.

Alleles↗
PubMed1997

Early cytokine and chemokine gene expression in lymph nodes of macaques infected with simian immunodeficiency virus is predictive of disease outcome and vaccine efficacy.

Competitive PCR was used to evaluate the expression of cytokine, granzyme B, and chemokine genes in lymph nodes of macaques recently infected with the simian immunodeficiency virus (SIV) pathogenic molecular clone SIVmac239 (n = 16), the nonpathogenic vaccine strain SIVmac239 delta nef (n = 8), and the nonpathogenic molecular clone SIVmac1A11 (n = 8). For both SIVmac239 and its nef-deleted derivative, strong expression was observed as early as 7 days postinfection for interleukin 1beta (IL-1beta), IL-6, tumor necrosis factor alpha, gamma interferon, and IL-13. The levels of gene induction were equally intense for both viruses despite a lower viral load for SIVmac239 deltanef compared with that for SIVmac239. However, the nature of the cytokine network activation varied with the viral inocula. Primary infection with SIVmac239 was characterized by a higher level of IL-4, IL-10, MIP-1alpha, MIP-1beta, MCP-1, and RANTES gene expression and a lower level of IL-12 and granzyme B gene expression compared with infection with SIVmac239 delta nef. Thus, infection with nef-deleted SIV was associated with a preferential Th1 versus Th2 pattern of cytokine production. Infection with SIVmac1A11 was characterized by a delayed immune response for all markers tested. The unique patterns of cytokine and chemokine gene expression in lymph nodes correlated nicely with the pathogenic potential of the SIV strains used as well as with differences in their ability to serve as protective vaccines.

Animals↗
PubMed1997

[The relationship between tumor necrosis factor and glomerular mesangial cells].

OBJECTIVE: To study the autocrine function of tumor necrosis factor (TNF) in glomerular mesangial cells (MC) and the action of TNF on MC. METHODS: Cultured rat and human MC were studied by RT-PCR, in situ hybridization and immunohistochemical methods. RESULTS: The expression of both TNF and TNF receptor mRNA and protein synthesis in MC were manifested. CONCLUSION: (1) MC is the target cell for TNF, MC have specific cell surface receptor to TNF. (2) TNF acts on MC through both autocrine and paracrine mechanism.

Animals↗