Structure, dynamics and immunogenicity of a catalytically inactive CXC chemokine-degrading protease SpyCEP from Streptococcus pyogenes

McKenna, Sophie; Malito, E.; Rouse, Sarah L.; Abate, Francesca; Bensi, Giuliano; Chiarot, Emiliano; Micoli, Francesca; Mancini, Francesca; Moriel, Danilo Gomes; Grandi, Guido; Mossakowska, Danuta E.; Pearson, Max; Xu, Yingqi; Pease, James E.; Sriskandan, Shiranee; Margarit, Immaculada; Bottomley, Matthew J.; Matthews, Stephen

doi:10.1016/j.csbj.2020.03.004

Cited by 19 publications

(30 citation statements)

References 63 publications

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“…In this case, the extent and location of the saccharide chains on the protein carrier might be relevant in terms of preservation of its key B-cell epitopes [ 21 ]. Moreover our results seem to confirm the importance of GAS protein antigens conformation for immunogenicity [ 40 , 41 ], as conjugation impacted on protein folding, as evidenced through DSC analysis, which was probably correlated with the loss of the functionality observed. Alternative conjugation methods, including protein site-selective chemical or enzymatic approaches [ 42 ] or bioconjugation [ 43 ], could be investigated to preserve protein protective epitopes, without impacting too much on protein structures and conformation.…”

Section: Discussionsupporting

confidence: 78%

Rational Design of a Glycoconjugate Vaccine against Group A Streptococcus

Benedetto

Mancini

Carducci

et al. 2020

IJMS

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No commercial vaccine is yet available against Group A Streptococcus (GAS), major cause of pharyngitis and impetigo, with a high frequency of serious sequelae in low- and middle-income countries. Group A Carbohydrate (GAC), conjugated to an appropriate carrier protein, has been proposed as an attractive vaccine candidate. Here, we explored the possibility to use GAS Streptolysin O (SLO), SpyCEP and SpyAD protein antigens with dual role of antigen and carrier, to enhance the efficacy of the final vaccine and reduce its complexity. All protein antigens resulted good carrier for GAC, inducing similar anti-GAC IgG response to the more traditional CRM197 conjugate in mice. However, conjugation to the polysaccharide had a negative impact on the anti-protein responses, especially in terms of functionality as evaluated by an IL-8 cleavage assay for SpyCEP and a hemolysis assay for SLO. After selecting CRM197 as carrier, optimal conditions for its conjugation to GAC were identified through a Design of Experiment approach, improving process robustness and yield This work supports the development of a vaccine against GAS and shows how novel statistical tools and recent advancements in the field of conjugation can lead to improved design of glycoconjugate vaccines.

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Section: Discussionsupporting

confidence: 78%

Rational Design of a Glycoconjugate Vaccine against Group A Streptococcus

Benedetto

Mancini

Carducci

et al. 2020

IJMS

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“…Several prospective antigens have been shown to be protective and effective vaccine candidates despite low OPA titres, such as SpyCEP [ 15 , 24 ]. SpyCEP and Mac immune sera are unlikely to show protective activity in an assay such as an OPA due to their localisation and the functional protease activity of the antigens.…”

Section: Resultsmentioning

confidence: 99%

“…Several prospective antigens have been shown to be protective and effective vaccine candidates despite low OPA titres, such as SpyCEP [15,24]. SpyCEP and Mac immune Figure 2.…”

Section: Immune Sera Can Block the Functional Activity Of Secreted Gas Virulence Factorsmentioning

confidence: 99%

“…In the present study, we have prepared and evaluated four recombinant protein antigens, in addition to M-proteins, particularly to be representative of different stages of GAS infection: Cpa, a pilin tip protein which caps the pilus polymer has been shown to display adhesion properties in some serotypes, hypothesised to play a role in GAS colonisation [ 16 , 17 ]; MalE, a cell associated maltodextrin binding protein essential for GAS survival in saliva [ 18 , 19 ]; Mac-1/IdeS (henceforth referred to as Mac), a secreted IgG protease that plays a role in the evasion of immune mechanisms by preventing opsonisation and phagocytosis [ 20 , 21 , 22 ] and SpyCEP, a cell surface and secreted IL-8 protease that reduces the recruitment of neutrophils, allowing GAS to evade immune clearance [ 23 , 24 ] and additionally playing a role in the invasion of cells and dissemination of GAS [ 25 , 26 ]. SpyCEP is included in the GSK combo vaccine and Combo5 and has been shown to be protective against infection in several preclinical studies [ 25 , 26 ], while the remaining three antigens are novel to vaccine evaluations.…”

Section: Introductionmentioning

confidence: 99%

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Multicomponent Vaccines against Group A Streptococcus Can Effectively Target Broad Disease Presentations

et al. 2021

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Group A Streptococcus (GAS) is an important global human pathogen, with a wide range of disease presentations, from mild mucosal infections like pharyngitis to invasive diseases such as toxic shock syndrome. The effect on health and mortality from GAS infections is substantial worldwide, particularly from autoimmune sequelae-like rheumatic heart disease (RHD), and there is currently no licenced vaccine. We investigated protein antigens targeting a broad range of GAS disease presentations as vaccine components in individual and combination formulations. The potency and functional immunity generated were evaluated and compared between groups. Antibodies against all components were found in pooled human IgG (IVIG) and an immune response generated following the subcutaneous immunisation of mice. A combination immunisation showed a reduction in IgG response for SpyCEP but an increase for Cpa and Mac-1 (IdeS). An opsonophagocytosis assay (OPA) showed the killing of GAS with immune sera against M protein and combination groups, with a lower killing activity observed for immune sera against other individual antigens. Specific antigen assays showed functional immunity against SpyCEP and Mac-1 from both individual and combination immunisations, with the activity correlating with antibody titres. However, efficient blocking of the binding activity of Cpa to collagen I and fibronectin could not be demonstrated with immune sera or purified IgG. Our data indicate that combination immunisations, while effective at covering a broader range of virulence factors, can also affect the immune response generated. Further, our results showed that an OPA alone is inadequate for understanding protection from vaccination, particularly when considering protection from immune evasion factors and evaluation of the colonisation leading to pharyngitis.

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“…Indeed, invasive GAS infections are estimated to cause more than 18 million cases worldwide leading to 517,000 deaths and an approximated 1.78 million new cases per year ( 8 ). As there is no vaccine against this pathogen ( 9 ), therapies need to be improved which in turn requires a better understanding of the pathological manifestations leading to severe disease courses like streptococcal toxic shock syndrome, necrotizing fasciitis, and septic arthritis (SA).…”

Section: Introductionmentioning

confidence: 99%

Inflammatory Joint Disease Is a Risk Factor for Streptococcal Sepsis and Septic Arthritis in Mice

et al. 2020

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Septic arthritis is a medical emergency associated with high morbidity and mortality, yet hardly any novel advances exist for its clinical management. Despite septic arthritis being a global health burden, experimental data uncovering its etiopathogenesis remain scarce. In particular, any interplay between septic arthritis and preceding joint diseases are unknown as is the contribution of the synovial membrane to the onset of inflammation. Using C57BL/6 mice as a model to study sepsis, we discovered that Group A Streptococcus (GAS) – an important pathogen causing septic arthritis - was able to invade the articular microenvironment. Bacterial invasion resulted in the infiltration of immune cells and detrimental inflammation. In vitro infected fibroblast-like synoviocytes induced the expression of chemokines ( Ccl2 , Cxcl2 ), inflammatory cytokines ( Tnf , Il6 ), and integrin ligands (ICAM-1, VCAM-1). Apart from orchestrating immune cell attraction and retention, synoviocytes also upregulated mediators impacting on bone remodeling ( Rankl ) and cartilage integrity ( Mmp13 ). Using collagen-induced arthritis in DBA/1 × B10.Q F1 mice, we could show that an inflammatory joint disease exacerbated subsequent septic arthritis which was associated with an excessive release of cytokines and eicosanoids. Importantly, the severity of joint inflammation controlled the extent of bone erosions during septic arthritis. In order to ameliorate septic arthritis, our results suggest that targeting synoviocytes might be a promising approach when treating patients with inflammatory joint disease for sepsis.

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Structure, dynamics and immunogenicity of a catalytically inactive CXC chemokine-degrading protease SpyCEP from Streptococcus pyogenes

Cited by 19 publications

References 63 publications

Rational Design of a Glycoconjugate Vaccine against Group A Streptococcus

Rational Design of a Glycoconjugate Vaccine against Group A Streptococcus

Multicomponent Vaccines against Group A Streptococcus Can Effectively Target Broad Disease Presentations

Inflammatory Joint Disease Is a Risk Factor for Streptococcal Sepsis and Septic Arthritis in Mice

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