Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation

Woodward, Michael P.; Young, William W.; Bloodgood, Robert A.

doi:10.1016/0022-1759(85)90337-0

Cited by 706 publications

(430 citation statements)

References 19 publications

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“…Modifications carried out on cell surface carbohydrates, therefore, are likely to influence the adhesive properties of cell surfaces. Periodate oxidation at acid pH has long been used as a tool to cleave carbohydrate residues (Woodward, Young & Bloodgood, 1985). The results presented here after the use of a whole range of periodate concentrations at low and neutral pH, show that carbohydrates are likely to be involved in T. congolense binding to endothelial cells.…”

Section: Discussionmentioning

confidence: 67%

The interaction ofTrypanosoma congolensewith endothelial cells

1994

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SUMMARYFactors which affect adhesion of cultured Trypanosoma congolense bloodstream forms to mammalian feeder cells have been examined. Using an in vitro binding assay, the initial events following interaction of trypanosomes with bovine aorta endothelial (BAE) cells were monitored by both light-and electron microscopy. Metabolic inhibitors and other biochemicals were incubated with either cells or parasites, to test whether any inhibited the process. Our findings suggest that adhesion of the parasites is an active process requiring metabolic energy from the trypanosomes, but not from endothelial cells. We also provide data suggesting that T. congolense bloodstream forms possess a lectin-like domain, localized at distinct sites on their flagellar surface, which interacts with specific carbohydrate receptors, most likely sialic acid residues, on the endothelial cell plasma membrane. We also suggest that the cytoskeletal protein actin is probably involved in this interaction.

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Section: Discussionmentioning

confidence: 67%

The interaction ofTrypanosoma congolensewith endothelial cells

1994

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“…Following the method of Woodward et al (1985) modified by Alarcón de Noya et al (2000), we incubated the blocked nitrocellulose strips in 10 mM SMP. After this treatment, tested sera were diluted 1:100 in blocking solution.…”

Section: Methodsmentioning

confidence: 99%

Antigenic community between Schistosoma mansoni and Biomphalaria glabrata: on the search of candidate antigens for vaccines

Chacón

Losada

Noya

et al. 2002

Mem. Inst. Oswaldo Cruz

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AWA (16, 30, 36, 60 and 155 kDa (Dissous et al. 1986, Iwanaga 1994, Weston et al. 1994, Gamal-Eddin et al. 1996, 1997. We demonstrated that sera from schistosome-infected persons reacted against soluble crude Biomphalaria glabrata antigen (SBgA) by ELISA (100% of sensitivity) (Alarcón de Noya et al. 1989) and that sera from mice immunized with SBgA recognized several homologous snail molecules by Western-blot .Cross-reactive antigens may probably result from the adaptation of parasites to their invertebrate and vertebrate hosts and in consequence, could prevent immune recognition in the latter. Our basic hypothesis is that the parasite acquires snail molecules on its surface that would cover critical antigens, allowing its survival during skin penetration and the evasion from the protective immune mechanisms of definitive host during the first events of invasion. In the present work, non-singenic mice were immunized with a crude antigen of non-infected B. glabrata in order to demonstrate, first, the cross-reactivity and crossinhibition between S. mansoni and B. glabrata common proteins. Second, to characterize SBgA, particularly its glycoprotein components and third, to determine the protective effect of the crude preparation of B. glabrata in mice against S. mansoni infection.

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“…Host cell-free HGE agent was incubated with 20 mM sodium periodate (Sigma) in 50 mM sodium acetate buffer (pH 4.5) at room temperature for 1 h in the dark as described by Woodward et al (25). Following a brief rinse with 50 mM sodium acetate, ehrlichiae were incubated with 50 mM sodium borohydride (Sigma) in phosphate-buffered saline at room temperature for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Human Granulocytic Ehrlichiosis Agent Inhibits Superoxide Anion Generation by Human Neutrophils

Mott

Rikihisa

2000

Infect Immun

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The human granulocytic ehrlichiosis (HGE) agent, which replicates in neutrophils, was found not to induce superoxide anion (O 2 ؊ ) generation or extracellular release by human peripheral blood neutrophils, as measured by a luminol-dependent chemiluminescence assay or a cytochrome c reduction assay, respectively. Furthermore, the HGE agent completely prevented O 2 ؊ release by neutrophils upon stimulation with phorbol myristate acetate (PMA), formylmethionyl-leucyl-phenylalanine, or Escherichia coli. The inhibition was HGE agent dose dependent, required ehrlichial contact with the host cells, and was reversible upon removal of the extracellular HGE agent bound to the host cells prior to PMA stimulation. Structural integrity of or new protein synthesis by the HGE agent was not required for the inhibition; carbohydrate but not surface protein of the HGE agent was required. The HGE agent did not prevent O 2 ؊ generation in human peripheral blood monocytes derived from the same individual. This neutrophil-specific prevention of O 2 ؊ generation by the HGE agent would be critical in survival of the HGE agent. This is the first demonstration of the rapid inhibition of preexisting NADPH oxidase in human neutrophils by the HGE agent.An emerging tick-borne zoonosis caused by the human granulocytic ehrlichiosis (HGE) agent was first described in 1994 (5) and has been increasingly recognized in the United States and several European countries. HGE is a disease characterized by systemic illness such as fever, chills, headache, malaise, and/or myalgia. Laboratory tests may reveal thrombocytopenia, leukopenia, elevated C-reactive protein levels, and elevated liver enzyme activities. The HGE agent is a unique obligatory intracellular bacterium that replicates in neutrophils. Neutrophils have a strong ability to kill invading microorganisms through activation of the NADPH oxidase, which rapidly generates superoxide anion (O 2 Ϫ ) and subsequent formation of other bactericidal reactive oxygen intermediates (hydrogen peroxide, myeloperoxide, hypochlorous acid, hydroxyl radical, or longer-lived N-chloroamines) (1, 7, 9). Recently Banerjee et al. (4) reported that after 5 days of infection with the HGE agent, HL-60 cells (a human promyelocytic leukemia cell line) exhibited a lack of luminol-dependent chemiluminescence (LDCL) response to phorbol myristate acetate (PMA) and down regulation of mRNA of gp91 phox , one component of the NADPH complex. However, for the HGE agent to survive in neutrophils, this inhibition must occur immediately rather than 5 days after establishment of infection. Although use of neutrophils is more difficult than use of cell lines, we determined whether human peripheral blood neutrophils produce O 2 Ϫ upon exposure to the HGE agent. Since we found that the HGE agent does not induce O 2 Ϫ generation, we determined whether this is a generalized and/or active inhibition and whether both intracellular and extracellular O 2 Ϫ generation is prevented. We further examined what type of ehrlichial factors and inter...

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Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation

Cited by 706 publications

References 19 publications

The interaction ofTrypanosoma congolensewith endothelial cells

The interaction ofTrypanosoma congolensewith endothelial cells

Antigenic community between Schistosoma mansoni and Biomphalaria glabrata: on the search of candidate antigens for vaccines

Human Granulocytic Ehrlichiosis Agent Inhibits Superoxide Anion Generation by Human Neutrophils

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