Colorimetric determination of neutral sugars by a resorcinol sulfuric acid micromethod

Monsigny, Michel; Petit, Claire; Roche, Annie‐Claude

doi:10.1016/0003-2697(88)90578-7

Cited by 341 publications

(188 citation statements)

References 6 publications

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“…Unreacted mannoside and polylysine were removed by centrifugal filtration through a dialysis membrane filter (Durapore TM CL3K, Millipore Corp.) at 5000 ϫ g for 30 min. The degree of sugar substitution was determined by the resorcinol sulfuric assay (19). The percentage of sugar in MPL was determined using the equation…”

Section: Methodsmentioning

confidence: 99%

Antisense Inhibition of Silica-induced Tumor Necrosis Factor in Alveolar Macrophages

Rojanasakul¹,

Weissman

Shi

et al. 1997

Journal of Biological Chemistry

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Tumor necrosis factor-␣ (TNF␣) has been shown to play an important role in the pathogenesis of silicotic fibrosis. In this study, antisense oligonucleotides targeted to TNF␣ mRNA were used to inhibit silica-induced TNF␣ gene expression in alveolar macrophages. To achieve macrophage-specific oligonucleotide delivery, a molecular conjugate consisting of mannosylated polylysine that exploits endocytosis via the macrophage mannose receptor was used. Complexes were formed between the mannosylated polylysine and oligonucleotides and added to the cells in the presence of silica. Enzyme-linked immunoadsorbent assay showed that the complex consisting of the conjugate and antisense oligomer effectively inhibited TNF␣ production, whereas the oligomer alone had much less effect. Reverse transcriptase-polymerase chain reaction analysis revealed that the reduction in TNF␣ secretion was associated with specific ablation of targeted TNF␣ mRNA. The conjugate alone or conjugate complexed with inverted or sense sequence oligonucleotide had no effect. The promoting effect of the conjugate on antisense activity was shown to be due to enhanced cellular uptake of the oligomer via mannose receptor-mediated endocytosis. Cells lacking mannose receptors showed no susceptibility to the conjugate treatment. These results indicate that effective and selective inhibition of macrophage TNF␣ expression can be achieved using the antisense mannosylated polylysine system.

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Section: Methodsmentioning

confidence: 99%

Antisense Inhibition of Silica-induced Tumor Necrosis Factor in Alveolar Macrophages

Rojanasakul¹,

Weissman

Shi

et al. 1997

Journal of Biological Chemistry

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“…The solution was mixed on the rotating wheel for 16 h at room temperature and then dialyzed overnight into PBS with 6000-8000 molecular weight cut-off (MWCO) tubing at 4 C. Through dialysis, unconjugated a-D-mannopyranosylphenyl isothiocyanate (MW 313.3) was removed. Mannose residues present in dendrimer were quantitated by a colorimetric assay as previously described [41]. To prepare MDO conjugates, 50 lL 0.2 M TCEP (Pierce, Gainesville, FL) in PBS was added to the MD solution and left for 30 min at room temperature to generate the reduced form of MD.…”

Section: Preparation Of Ova MD Mdo and Fitc Conjugatesmentioning

confidence: 99%

Delivery of antigen using a novel mannosylated dendrimer potentiates immunogenicity in vitro and in vivo

et al. 2008

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Antigen mannosylation has been shown to be an effective approach to potentiate antigen immunogenicity, due to the enhanced antigen uptake and presentation by APC. To overcome disadvantages associated with conventional methods used to mannosylate antigens, we have developed a novel mannose-based antigen delivery system that utilizes a polyamidoamine (PAMAM) dendrimer. It is demonstrated that mannosylated dendrimer ovalbumin (MDO) is a potent immune inducer. With a strong binding avidity to DC, MDO potently induced OVA-specific T cell response in vitro. It was found that the immunogenicity of MDO was due not only to enhanced antigen presentation, but also to induction of DC maturation. Mice immunized with MDO generated strong OVA-specific CD4 + /CD8 + T cell and antibody responses. MDO also targeted lymph node DC to crosspresent OVA, leading to OTI CD8 + T cell proliferation. Moreover, upon challenge with B16-OVA tumor cells, tumors in mice pre-immunized with MDO either did not grow or displayed a much more delayed onset, and had slower kinetics of growth than those of OVA-immunized mice. This mannose-based antigen delivery system was applied here for the first time to the immunization study. With several advantages and exceptional adjuvanticity, we propose mannosylated dendrimer as a potential vaccine carrier.

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“…The average number of glycosyl residues per PEI molecule was determined using the resorcinol/sulfuric acid micromethod. 21 The extent of grafting was expressed as the percentage of glycosyl residues per amino group of PEI and was found to be 5%. The polymer concentration was determined by the trinitrobenzene sulfonic acid assay for B-PEI and by the ninhydrin assay for L-PEI.…”

Section: Preparation Of Glycosylated Pei Conjugatesmentioning

confidence: 99%

Ovarian carcinoma cells are effectively transfected by polyethylenimine (PEI) derivatives

Poulain

Ziller

Muller³

et al. 2000

Cancer Gene Ther

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As a prerequisite to nonviral gene therapy approaches of ovarian carcinoma, we evaluated the possibility of transfecting established tumor cell lines (SKOV3, IGROV1) as well as primary mesothelial and tumor cells by various polyethylenimine (PEI) derivatives. Several PEI-based vectors were able to effectively transfect these cells, as shown by high luciferase expression levels (10 8 to 10 9 relative light units per milligram of cell protein) that corresponded with 25-50% of green fluorescent protein-positive cells after 24 hours. However, unpredictable differences were observed among the vectors and cell types that a posteriori justified the screening procedure. We also showed that cells that were not transfected after the first experiment remained transfectable in a subsequent transfection experiment to a level similar to that of the initial population. This experiment does not support the emergence of a transfection-resistant cell population and opens the door to multiple therapeutic gene deliveries. Although efficacy and cell targeting still remain to be improved, PEI derivatives appear to be promising molecules for the development of nonviral gene therapy of ovarian carcinoma. Cancer Gene Therapy (2000) 7, 644 -652

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Colorimetric determination of neutral sugars by a resorcinol sulfuric acid micromethod

Cited by 341 publications

References 6 publications

Antisense Inhibition of Silica-induced Tumor Necrosis Factor in Alveolar Macrophages

Antisense Inhibition of Silica-induced Tumor Necrosis Factor in Alveolar Macrophages

Delivery of antigen using a novel mannosylated dendrimer potentiates immunogenicity in vitro and in vivo

Ovarian carcinoma cells are effectively transfected by polyethylenimine (PEI) derivatives

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