scholarly journals Pathogenesis of shigella diarrhea. XI. Isolation of a shigella toxin-binding glycolipid from rabbit jejunum and HeLa cells and its identification as globotriaosylceramide.

1986 ◽  
Vol 163 (6) ◽  
pp. 1391-1404 ◽  
Author(s):  
M Jacewicz ◽  
H Clausen ◽  
E Nudelman ◽  
A Donohue-Rolfe ◽  
G T Keusch
Keyword(s):  
Hela Cells ◽  
Neutralizing Antibodies ◽  
Specific Binding ◽  
Jejunal Mucosa ◽  
Toxin B ◽  
Toxin Binding ◽  
B Subunit ◽  
Specific Binding Sites ◽  
Rabbit Jejunum ◽  
Group B

A glycolipid that specifically binds shigella toxin was isolated from both HeLa cells and rabbit jejunal mucosa and identified as globotriaosylceramide (Gb3) by its identical mobility on HPTLC to authentic erythrocyte Gb3. Toxin also bound to a band tentatively identified as alpha-hydroxylated Gb3. In addition, toxin bound to P1 antigen present in group B human erythrocyte glycolipid extracts. The common feature of the three binding glycolipids is a terminal Gal alpha 1----4Gal disaccharide linked beta 1----4 to either Glc or GlcNAc. Globoisotriaosylceramide, which differs from Gb3 only in possessing a Gal alpha 1----3Gal terminal disaccharide, and LacCer, which lacks the terminal Gal residue of Gb3, were incapable of binding the toxin. Binding was shown to be mediated by the B subunit by the use of isolated toxin A and B subunits and monoclonal subunit-specific antibodies. Gb3-containing liposomes competitively inhibited the binding of toxin to HeLa cell monolayers but did not inhibit toxin-induced cytotoxicity. These studies show an identical carbohydrate-specific glycolipid receptor for shigella toxin in gut and in HeLa cells. The toxin B subunit that mediates this binding has also been shown to recognize a glycoprotein receptor with different sugar specificity. Thus, we have demonstrated that the same small (Mr 6,500) B subunit polypeptide has two distinctive carbohydrate-specific binding sites. The Gal alpha 1----4Gal disaccharide of the glycolipid toxin receptor is also recognized by the Gal-Gal pilus of uropathogenic E. coli. This suggests the possibility that the pilus and toxin B subunit contain homologous sequences. If this is true, it may be possible to use the purified Gal-Gal pilus to produce toxin-neutralizing antibodies.

scholarly journals Shiga toxin–binding site for host cell receptor GPP130 reveals unexpected divergence in toxin-trafficking mechanisms

2013 ◽  
Vol 24 (15) ◽  
pp. 2311-2318 ◽  
Author(s):  
Somshuvra Mukhopadhyay ◽  
Brendan Redler ◽  
Adam D. Linstedt
Keyword(s):  
Shiga Toxin ◽  
Transmembrane Domain ◽  
Cell Receptor ◽  
Toxin B ◽  
Toxin Binding ◽  
B Subunit ◽  
Host Cell Receptor ◽  
Developed World ◽  
Trafficking Receptor ◽  
Shed Light

Shiga toxicosis is caused by retrograde trafficking of one of three types of Shiga toxin (STx), STx, STx1, or STx2. Trafficking depends on the toxin B subunits, which for STx and STx1 are identical and bind GPP130, a manganese (Mn)-sensitive intracellular trafficking receptor. Elevated Mn down-regulates GPP130, rendering STx/STx1 harmless. Its effectiveness against STx2, however, which is a serious concern in the developed world, is not known. Here we show that Mn-induced GPP130 down-regulation fails to block STx2 trafficking. To shed light on this result, we tested the purified B subunit of STx2 for binding to GPP130 and found that it failed to interact. We then mapped residues at the interface of the GPP130-STx/STx1 complex. In GPP130, binding mapped to a seven-residue stretch in its lumenal stem domain next to the transmembrane domain. This stretch was required for STx/STx1 transport. In STx/STx1, binding mapped to a histidine–asparagine pair on a surface-exposed loop of the toxin B subunit. Significantly, these residues are not conserved in STx2, explaining the lack of effectiveness of Mn against STx2. Together our results imply that STx2 uses an evolutionarily distinct trafficking mechanism and that Mn as a potential therapy should be focused on STx/STx1 outbreaks, which account for the vast majority of cases worldwide.

scholarly journals A Replicating Single-Cycle Adenovirus Vaccine Effective against Clostridium difficile

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 470
Author(s):  
William E. Matchett ◽  
Stephanie Anguiano-Zarate ◽  
Goda Baddage Rakitha Malewana ◽  
Haley Mudrick ◽  
Melissa Weldy ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Neutralizing Antibodies ◽  
Antibody Responses ◽  
Complete Protection ◽  
Single Cycle ◽  
Toxin B ◽  
Toxin Binding ◽  
Binding Domains ◽  
Binding Antibody

Clostridium difficile causes nearly 500,000 infections and nearly 30,000 deaths each year in the U.S., which is estimated to cost $4.8 billion. C. difficile infection (CDI) arises from bacteria colonizing the large intestine and releasing two toxins, toxin A (TcdA) and toxin B (TcdB). Generating humoral immunity against C. difficile’s toxins provides protection against primary infection and recurrence. Thus, a vaccine may offer the best opportunity for sustained, long-term protection. We developed a novel single-cycle adenovirus (SC-Ad) vaccine against C. difficile expressing the receptor-binding domains from TcdA and TcdB. The single immunization of mice generated sustained toxin-binding antibody responses and protected them from lethal toxin challenge for up to 38 weeks. Immunized Syrian hamsters produced significant toxin-neutralizing antibodies that increased over 36 weeks. Single intramuscular immunization provided complete protection against lethal BI/NAP1/027 spore challenge 45 weeks later. These data suggest that this replicating vaccine may prove useful against CDI in humans.

scholarly journals Intranasal Immunization of Mice with Group B Streptococcal Protein Rib and Cholera Toxin B Subunit Confers Protection against Lethal Infection

2004 ◽  
Vol 72 (2) ◽  
pp. 1184-1187 ◽  
Author(s):  
Charlotte Larsson ◽  
Jan Holmgren ◽  
Gunnar Lindahl ◽  
Charlotta Bergquist
Keyword(s):  
Cholera Toxin ◽  
Group B Streptococcus ◽  
Surface Protein ◽  
Cholera Toxin B Subunit ◽  
Toxin B ◽  
Intranasal Immunization ◽  
Cholera Toxin B ◽  
B Subunit ◽  
A Cell ◽  
Group B

ABSTRACT Intranasal immunization of mice with Rib, a cell surface protein of group B streptococcus (GBS), conjugated to or simply coadministered with the recombinant cholera toxin B subunit, induces systemic immunoglobulin G (IgG) and local IgA antibody responses and confers protection against lethal GBS infection. These findings have implications for the development of a human GBS vaccine.

scholarly journals Oral immunization with a synthetic peptide of cholera toxin B subunit. Obtention of neutralizing antibodies

1986 ◽  
Vol 159 (3) ◽  
pp. 525-528 ◽  
Author(s):  
Anne GUYON-GRUAZ ◽  
Agnes DELMAS ◽  
Sylvie PEDOUSSAUT ◽  
Hubert HALIMI ◽  
Gerard MILHAUD ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Synthetic Peptide ◽  
Neutralizing Antibodies ◽  
Oral Immunization ◽  
Cholera Toxin B Subunit ◽  
Toxin B ◽  
Cholera Toxin B ◽  
B Subunit

scholarly journals Group B Streptococcus Capsular Polysaccharide-Cholera Toxin B Subunit Conjugate Vaccines Prepared by Different Methods for Intranasal Immunization

2001 ◽  
Vol 69 (1) ◽  
pp. 297-306 ◽  
Author(s):  
Xuzhuang Shen ◽  
Teresa Lagergård ◽  
Yonghong Yang ◽  
Marianne Lindblad ◽  
Margareta Fredriksson ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Capsular Polysaccharide ◽  
Cholera Toxin B Subunit ◽  
Small Molecular Weight ◽  
Toxin B ◽  
Conjugate Vaccines ◽  
Intranasal Immunization ◽  
Cholera Toxin B ◽  
B Subunit ◽  
Group B

ABSTRACT Group B Streptococcus (GBS) type III capsular polysaccharide (CPS III) was conjugated to recombinant cholera toxin B subunit (rCTB) using three different methods which employed (i) cystamine and N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), (ii) carbodiimide with adipic acid dihydrazide (ADH) as a spacer, or (iii) reductive amination (RA). The CPS III-rCTB conjugates were divided into large- and small-molecular-weight (M r) fractions, and the immunogenicities of the different preparations after intranasal (i.n.) immunization were studied in mice. Both large- and small-M rconjugates of CPS III-rCTBRA or CPS III-rCTBADHinduced high, almost comparable levels of CPS-specific immunoglobulin G (IgG) in serum, lungs, and vagina that were generally superior to those obtained with CPS III-rCTBSPDP conjugates or a CPS III and rCTB mixture. However, the smaller-M rconjugates of CPS III-rCTBRA or CPS III-rCTBADHin most cases elicited a lower anti-CPS IgA immune response than the large-M r conjugates, and the highest anti-CPS IgA titers in both tissues and serum were obtained with the large-M r CPS III-rCTBRA conjugate. Serum IgG anti-CPS titers induced by the CPS III-rCTBRAconjugate had high levels of specific IgG1, IgG2a, IgG2b, and IgG3 antibodies. Based on the effectiveness of RA for coupling CPS III to rCTB, RA was also tested for conjugating GBS CPS Ia with rCTB. As for the CPS III-rCTB conjugates, the immunogenicity of CPS Ia was greatly increased by conjugation to rCTB. Intranasal immunization with a combination of CPS Ia-rCTB and CPS III-rCTB conjugates was shown to induce anti-CPS Ia and III immune responses in serum and lungs that were fully comparable with the responses to immunization with the monovalent CPS Ia-rCTB or CPS III-rCTB conjugates. These results suggest that the GBS CPS III-rCTB and CPS Ia-rCTB conjugates prepared by the RA method may be used in bivalent and possibly also in multivalent mucosal GBS conjugate vaccines.

Oral immunization with a lettuce-derived Escherichia coli heat-labile toxin B subunit induces neutralizing antibodies in mice

2011 ◽  
Vol 107 (3) ◽  
pp. 441-449 ◽  
Author(s):  
Luzmila Martínez-González ◽  
Sergio Rosales-Mendoza ◽  
Ruth Elena Soria-Guerra ◽  
Leticia Moreno-Fierros ◽  
Rubén López-Revilla ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Neutralizing Antibodies ◽  
Oral Immunization ◽  
Toxin B ◽  
B Subunit ◽  
Heat Labile

Targeting of Shiga Toxin B-Subunit to Retrograde Transport Route in Association with Detergent-resistant Membranes

2001 ◽  
Vol 12 (8) ◽  
pp. 2453-2468 ◽  
Author(s):  
Thomas Falguières ◽  
Frédéric Mallard ◽  
Carole Baron ◽  
Daniel Hanau ◽  
Clifford Lingwood ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Shiga Toxin ◽  
Secretory Pathway ◽  
Retrograde Transport ◽  
Toxin B ◽  
B Subunit ◽  
Early Endosomes ◽  
Golgi Network ◽  
Triton X

In HeLa cells, Shiga toxin B-subunit is transported from the plasma membrane to the endoplasmic reticulum, via early endosomes and the Golgi apparatus, circumventing the late endocytic pathway. We describe here that in cells derived from human monocytes, i.e., macrophages and dendritic cells, the B-subunit was internalized in a receptor-dependent manner, but retrograde transport to the biosynthetic/secretory pathway did not occur and part of the internalized protein was degraded in lysosomes. These differences correlated with the observation that the B-subunit associated with Triton X-100-resistant membranes in HeLa cells, but not in monocyte-derived cells, suggesting that retrograde targeting to the biosynthetic/secretory pathway required association with specialized microdomains of biological membranes. In agreement with this hypothesis we found that in HeLa cells, the B-subunit resisted extraction by Triton X-100 until its arrival in the target compartments of the retrograde pathway, i.e., the Golgi apparatus and the endoplasmic reticulum. Furthermore, destabilization of Triton X-100-resistant membranes by cholesterol extraction potently inhibited B-subunit transport from early endosomes to thetrans-Golgi network, whereas under the same conditions, recycling of transferrin was not affected. Our data thus provide first evidence for a role of lipid asymmetry in membrane sorting at the interface between early endosomes and the trans-Golgi network.

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