Biochemical and biological characteristics of cross-reacting material 197 (CRM197), a non-toxic mutant of diphtheria toxin: Use as a conjugation protein in vaccines and other potential clinical applications

Bröker, Michael; Costantino, Paolo; DeTora, Lisa; McIntosh, E. David G.; Rappuoli, Rino

doi:10.1016/j.biologicals.2011.05.004

Cited by 110 publications

(104 citation statements)

References 64 publications

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“…The effect of varying such parameters is antigen dependent and was investigated in the current study to optimize the design of a vaccine against MenX. Three different conjugate vaccines were synthesized using MenX OS of avDP 15-20 or 80-100 and CRM 197 (34) as carrier protein. The chemistry developed for Menveo was adapted to the shorter OS (35).…”

Section: Discussionmentioning

confidence: 99%

“…MenX oligosaccharides (OS) of different length and three different conjugation chemistries were compared, using CRM 197, a nontoxic mutant of diphtheria toxin (34), as carrier protein. When tested in mice, all MenX-CRM 197 conjugates resulted in high IgG antibody levels and serum bactericidal activity (SBA) titers, indicating the path ahead for the clinical development of a vaccine to prevent the remaining cause of epidemic meningococcal meningitis in Africa for which no vaccine is available.…”

Section: Significancementioning

confidence: 99%

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Development of a glycoconjugate vaccine to prevent meningitis in Africa caused by meningococcal serogroup X

Micoli

Romano

Tontini

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Neisseria meningitidis is a major cause of bacterial meningitis worldwide, especially in the African meningitis belt, and has a high associated mortality. The meningococcal serogroups A, W, and X have been responsible for epidemics and almost all cases of meningococcal meningitis in the meningitis belt over the past 12 y. Currently no vaccine is available against meningococcal X (MenX). Because the development of a new vaccine through to licensure takes many years, this leaves Africa vulnerable to new epidemics of MenX meningitis at a time when the epidemiology of meningococcal meningitis on the continent is changing rapidly, following the recent introduction of a glycoconjugate vaccine against serogroup A. Here, we report the development of candidate glycoconjugate vaccines against MenX and preclinical data from their use in animal studies. Following optimization of growth conditions of our seed MenX strain for polysaccharide (PS) production, a scalable purification process was developed yielding high amounts of pure MenX PS. Different glycoconjugates were synthesized by coupling MenX oligosaccharides of varying chain length to CRM 197 as carrier protein. Analytical methods were developed for in-process control and determination of purity and consistency of the vaccines. All conjugates induced high anti-MenX PS IgG titers in mice. Antibodies were strongly bactericidal against African MenX isolates. These findings support the further development of glycoconjugate vaccines against MenX and their assessment in clinical trials to produce a vaccine against the one cause of epidemic meningococcal meningitis that currently cannot be prevented by available vaccines.

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Section: Discussionmentioning

confidence: 99%

Section: Significancementioning

confidence: 99%

Development of a glycoconjugate vaccine to prevent meningitis in Africa caused by meningococcal serogroup X

Micoli

Romano

Tontini

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…In the present study, we synthesized glycoconjugate vaccines against S. Typhimurium, with OAg linked to the nontoxic mutant of diphtheria toxin, CRM 197 (26). OAg was extracted by acetic acid hydrolysis directly from the bacterial pellet, a process that removes the toxic lipid A by cleaving the labile bond between lipid A and the KDO sugar at the end of the core region (27).…”

mentioning

confidence: 99%

Design of Glycoconjugate Vaccines against Invasive African Salmonella enterica Serovar Typhimurium

et al. 2015

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dNontyphoidal salmonellae, particularly Salmonella enterica serovar Typhimurium, are a major cause of invasive disease in Africa, affecting mainly young children and HIV-infected individuals. Glycoconjugate vaccines provide a safe and reliable strategy against invasive polysaccharide-encapsulated pathogens, and lipopolysaccharide (LPS) is a target of protective immune responses. With the aim of designing an effective vaccine against S. Typhimurium, we have synthesized different glycoconjugates, by linking O-antigen and core sugars (OAg) of LPS to the nontoxic mutant of diphtheria toxin (CRM 197 ). The OAg-CRM 197 conjugates varied in (i) OAg source, with three S. Typhimurium strains used for OAg extraction, producing OAg with differences in structural specificities, (ii) OAg chain length, and (iii) OAg/CRM 197 ratio. All glycoconjugates were compared for immunogenicity and ability to induce serum bactericidal activity in mice. In vivo enhancement of bacterial clearance was assessed for a selected S. Typhimurium glycoconjugate by challenge with live Salmonella. We found that the largest anti-OAg antibody responses were elicited by (i) vaccines synthesized from OAg with the highest glucosylation levels, (ii) OAg composed of mixed-or medium-molecular-weight populations, and (iii) a lower OAg/CRM 197 ratio. In addition, we found that bactericidal activity can be influenced by S. Typhimurium OAg strain, most likely as a result of differences in OAg O-acetylation and glucosylation. Finally, we confirmed that mice immunized with the selected OAg-conjugate were protected against S. Typhimurium colonization of the spleen and liver. In conclusion, our findings indicate that differences in the design of OAg-based glycoconjugate vaccines against invasive African S. Typhimurium can have profound effects on immunogenicity and therefore optimal vaccine design requires careful consideration.

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“…Therefore, to investigate whether DT is really endowed with HS-binding ability and whether this characteristic affects its T cell presentation, we decided to perform studies at pH 7.4 and pH 6. In these studies, we mainly used a structurally similar nontoxic mutant of DT, called CRM197, which differs from the parent protein by a single residue mutation located in the catalytic domain (25,26). In the present study, we describe the results of binding to purified HS-and HSPG-expressing cells and the effects of this binding on uptake as well as on MHC class II-restricted presentation to Th cells.…”

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confidence: 99%

Acidosis Increases MHC Class II–Restricted Presentation of a Protein Endowed with a pH-Dependent Heparan Sulfate–Binding Ability

Knittel

Savatier

Upert

et al. 2015

The Journal of Immunology

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Heparan sulfate proteoglycans (HSPGs) are ubiquitously expressed molecules that participate in numerous biological processes. We previously showed that HSPGs expressed on the surface of APCs can serve as receptors for a hybrid protein containing an HS ligand and an Ag, which leads to more efficient stimulation of Th cells. To investigate whether such behavior is shared by proteins with inherent HS-binding ability, we looked for proteins endowed with this characteristic. We found that diphtheria toxin and its nontoxic mutant, called CRM197, can interact with HS. However, we observed that their binding ability is higher at pH 6 than at pH 7.4. Therefore, as extracellular acidosis occurs during infection by various micro-organisms, we assessed whether HS-binding capacity affects MHC class II–restricted presentation at different pHs. We first observed that pH decrease allows CRM197 binding to HSPG-expressing cells, including APCs. Then, we showed that this interaction enhances Ag uptake and presentation to Th cells. Lastly, we observed that pH decrease does not affect processing and presentation abilities of the APCs. Our findings show that acidic pH causes an HSPG-mediated uptake and an enhancement of T cell stimulation of Ags with the inherent ability to bind HSPGs pH-dependently. Furthermore, they suggest that proteins from micro-organisms with this binding characteristic might be supported more efficiently by the adaptive immune system when acidosis is triggered during infection.

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Biochemical and biological characteristics of cross-reacting material 197 (CRM197), a non-toxic mutant of diphtheria toxin: Use as a conjugation protein in vaccines and other potential clinical applications

Cited by 110 publications

References 64 publications

Development of a glycoconjugate vaccine to prevent meningitis in Africa caused by meningococcal serogroup X

Development of a glycoconjugate vaccine to prevent meningitis in Africa caused by meningococcal serogroup X

Design of Glycoconjugate Vaccines against Invasive African Salmonella enterica Serovar Typhimurium

Acidosis Increases MHC Class II–Restricted Presentation of a Protein Endowed with a pH-Dependent Heparan Sulfate–Binding Ability

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