A Phase I Study of Chemically Synthesized Verotoxin (Shiga-like Toxin) Pk-Trisaccharide Receptors Attached to Chromosorb for Preventing Hemolytic-Uremic Syndrome

Armstrong, Glen D.; Rowe, Peter C.; Goodyer, Paul; Orrbine, Elaine; Klassen, Terry P; Wells, George A.; Mackenzie, A. M. R.; Lior, H.; Blanchard, Colline; Auclair, François; Thompson, Brad; Rafter, D. J.; McLaine, Peter N.

doi:10.1093/infdis/171.4.1042

Cited by 132 publications

(72 citation statements)

References 10 publications

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“…Theoretically, our findings would support the recently proposed antiadhesive therapies aimed at blocking the binding of Stx to its glycosphingolipid receptor. 62,63 BLOOD, 15 MAY 2001 ⅐ VOLUME 97, NUMBER 10 For personal use only. on May 11, 2018.…”

Section: Discussionmentioning

confidence: 99%

Platelet activation by Shiga toxin and circulatory factors as a pathogenetic mechanism in the hemolytic uremic syndrome

Karpman

Papadopoulou

Nilsson

et al. 2001

Blood

129

117

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Thrombocytopenia caused by platelet consumption in thrombi is a major manifestation of hemolytic uremic syndrome (HUS) associated with Shiga toxin (Stx) producing Escherichia coli. Platelets have glycosphingolipid receptors capable of binding Stx, but a direct interaction between the toxin and platelets, leading to platelet activation, has not been reported. In this study, it is shown that Stx1 and its B (binding) subunit (Stx1B), at 10 pg/mL to 10 ng/mL, bound to platelets. IntroductionShiga toxin (Stx)-producing Escherichia coli (STEC) have been associated with cases of hemolytic uremic syndrome (HUS). Microangiopathic hemolytic anemia, acute renal failure, and thrombocytopenia characterize this syndrome. Stx has been specifically implicated as a causal factor 1 of HUS because cases of the disease have been associated with Stx-producing strains, 2 the toxin has been identified in the kidney of a patient with HUS, 3 and the toxin has been found to be cytotoxic for renal endothelial and epithelial cells. [4][5][6][7][8] In addition, animal models have reproduced aspects of HUS using wild-type bacteria that produce the toxin 9,10 or purified toxin. 11 The toxin is composed of 5 B, or binding, subunits and 1 enzymatically active A subunit. 12 The molecular weight of the A subunit is 33 000 kd, and that of the pentameric B subunit is approximately 35 000 kd. 13 The B subunit (StxB) binds to a glycosphingolipid receptor on cell membranes, globotriaosyl ceramide (Gb3), containing the terminal disaccharide galactose ␣(1-4) galactose ␤. 14 A recent study showed that Stx could bind to platelets through Gb3 and a novel platelet glycosphingolipid termed band 0.03. 15 Other studies have not been able to reproduce these results. 16,17 Binding to the cell mediates endocytosis of the holotoxin. Intracellularly the A subunit is proteolytically nicked, binds to 60S ribosomes, and thereby inhibits protein synthesis leading to cell death. 18 Thrombocytopenia is an important feature of HUS. Platelets are consumed in small thrombi, and these thrombi may compromise the blood supply to various organs, particularly the kidney. The mechanism by which these thrombi are formed is unclear, and platelet activation has been suggested. Studies performed on platelets from patients with HUS showed impaired aggregating responses 19,20 and reduced ␤-thromboglobulin (␤-TG) content. 19 Platelet-derived products such as ␤-TG, platelet factor 4, 21 and soluble P-selectin 22 were found to be elevated during acute HUS. Furthermore, changes in platelet ultramorphology were noted. 23 Increased platelet-derived microvesicles were found in these patients, 24 indicating platelet activation. Plasma from patients with HUS increased aggregation of normal platelets, but the factor causing this was not determined. 25 Several previous studies have investigated a direct interaction between Stx and platelets. Culture filtrates from Stx-producing E coli were found to induce platelet-aggregating activity, 26 but purified Stx did not increase platelet aggregation 16...

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Section: Discussionmentioning

confidence: 99%

Platelet activation by Shiga toxin and circulatory factors as a pathogenetic mechanism in the hemolytic uremic syndrome

Karpman

Papadopoulou

Nilsson

et al. 2001

Blood

129

117

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“…MCO moiety is similar in length to a ceramide of Gb3. A recently completed double-blind, placebo-controlled phase I study in healthy adult volunteers has revealed the safety and stability of Synsorb Pk in the human gut (2).Further, a phase II study has been completed, and a phase III study is now ongoing (3). Information of the clinical trials prompted us to evaluate the potential of Synsorb Pk for use in patients with STEC infections in Japan.…”

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confidence: 99%

In Vitro Assessment of a Chemically Synthesized Shiga Toxin Receptor Analog Attached to Chromosorb P (Synsorb Pk) as a Specific Absorbing Agent of Shiga Toxin 1 and 2

Takeda¹,

Yoshino²,

Adachi³

et al. 1999

Microbiology and Immunology

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A synthetic analog of Shiga toxin (Stx) receptor (Synsorb Pk) was quantitatively assessed to determine whether it can protect human renal adenocarcinoma cells (ACHN cells) from the cytotoxicity of Stx1 and Stx2 by coincubation experiments. Coincubation of 100 and 20 ng of Stx1 and Stx2 with 50 mg of Synsorb Pk for 1 hr at 37 C in 1 ml of Eagle's Minimum Essential Medium supplemented with 1% (v/v) nonessential amino acid and 10% (v/v) fetal calf serum protected 50% of the cells from the cytotoxic effect. Chromosorb P, an inert matrix control, did not absorb the Stxs at all. Heat‐treatment (boiled for 10 min) to Synsorb Pk caused a 50% decrease in Stx2‐binding activity, but did not effect the Stx1 binding. Further, Stxs bound to Synsorb Pk could be demonstrated. When 20 mg of Synsorb Pk was coincubated for 30 min at 37 C in 1 ml of phosphate‐buffered saline with 1 and 10 ng or more of Stx1 or Stx2, respectively, the toxins could be detected on the surface when the bound toxins on Synsorb Pk were used as the solid phase in enzyme immunoassay. The amount of 100 ng/ml of both Stx1 and Stx2 appeared to saturate 20 mg/ml of Synsorb Pk after coincubating for 30 min at 37 C. While assessing the Stxs' binding activity to Synsorb Pk, it was demonstrated that Stx1 had a higher affinity to Pk trisaccharide than Stx2. These observations provide useful information on the effectiveness of Synsorb Pk to trap and eliminate free Stxs produced in the gut of patients infected by Stx‐producing Escherichia coli, and to prevent the progression of hemorrhagic colitis to hemolytic uremic syndrome.

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“…In the course of this meeting, the clinical, epidemiological and biological features of shigellosis and STEC infections will be thoroughly reviewed, highlighting the prospects for developing new control measures. With respect to the Shiga toxins there are already clinical trials underway to fully examine the potential of adsorption of toxin in the intestinal lumen using a nonabsorbable receptor analogue (Synsorb-pk) (62). This strategy aims at blocking toxin uptake to the systemic circulation by irreversibly binding toxin to a silica particle that will be excreted in the stool, however the toxin pool present in the bowel lumen may be of limited importance in pathogenesis, as the small amount of toxin needed for systemic manifestations may be absorbed directly at the site of the intimate attachment of the organisms to the mucosa.…”

Section: Introductionmentioning

confidence: 99%

The Rediscovery of Shiga Toxin and Its Role in Clinical Disease

Keusch¹

1998

JJMSB

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A Phase I Study of Chemically Synthesized Verotoxin (Shiga-like Toxin) Pk-Trisaccharide Receptors Attached to Chromosorb for Preventing Hemolytic-Uremic Syndrome

Cited by 132 publications

References 10 publications

Platelet activation by Shiga toxin and circulatory factors as a pathogenetic mechanism in the hemolytic uremic syndrome

Platelet activation by Shiga toxin and circulatory factors as a pathogenetic mechanism in the hemolytic uremic syndrome

In Vitro Assessment of a Chemically Synthesized Shiga Toxin Receptor Analog Attached to Chromosorb P (Synsorb Pk) as a Specific Absorbing Agent of Shiga Toxin 1 and 2

The Rediscovery of Shiga Toxin and Its Role in Clinical Disease

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