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New modular platform based on multi-adjuvanted amphiphilic chitosan nanoparticles for efficient lipopeptide vaccine delivery against group a streptococcus

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Asian Journal of Pharmaceutical Sciences 2022年第3期17卷 435-446页

作者： Abdin Shakirin Mohamad Norpi Muhammad Luqman Nordin Nuraziemah Ahmad Haliza Katas Abdullah Al-Hadi Ahmad Fuaad Asif Sukri Nirmal Marasini Fazren Azmi Centre for Drug Delivery Technology Faculty of PharmacyUniversiti Kebangsaan MalaysiaKuala Lumpur 50300Malaysia Faculty Pharmacy and Health Sciences Universiti Kuala LumpurRoyal College of Medicine PerakPerak 30450Malaysia Faculty of Veterinary Medicine Universiti Malaysia KelantanKelantan 16100Malaysia Department of Chemistry Faculty of ScienceUniversiti MalayaKuala Lumpur 50603Malaysia Universiti Teknologi Mara(UiTM) Bandar Puncak AlamIntegrative Pharmacogenomics Institute(iPROMISE)Selangor 43200Malaysia School of Biomedical Sciences The University of QueenslandSt.Lucia QLD 4072Australia

An effective vaccine against group a streptococcus(GAS)is highly desirable for definitive control of GAS infections.In the present study,two variants of amphiphilic chitosan nanoparticles-based GAS vaccines were developed.The vaccines were primarily composed of encapsulated KLH protein(a source of T helper cell epitopes)and lipidated M-protein derived B cell peptide epitope(lipoJ14)within the amphiphilic structure of nanoparticles.The only difference between themwas one of the nanoparticles vaccines received additional surface coating with poly(I:C).The formulated vaccines exhibited nanosized particles within the range of 220–240 nm.Cellular uptake study showed that nanoparticles vaccine without additional poly(I:C)coating has greater uptake by dendritic cells and macrophages compared to nanoparticles vaccine that was functionalized with poly(I:C).Both vaccines were found to be safe in mice and showed negligible cytotoxicity against HEK293 cells.Upon immunization in mice,both nanoparticle vaccines produced high antigen-specific antibodies titres that were regulated by a balanced Th1 and Th2 response compared to physical mixture.These antibodies elicited high opsonic activity against the tested GAS strains.Overall,our data demonstrated that amphiphilic chitosan nanoparticles platform induced a potent immune response even without additional inclusion of poly(I:C).

关键词： Amphiphilic chitosan nanoparticles Peptide vaccine Lipidation Multi-adjuvanting delivery system Immunogenicity group a streptococcus

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The group a streptococcus Interleukin-8 Protease SpyCEP Promotes Bacterial Intracellular Survival by Evasion of Autophagy

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Infectious Microbes & Diseases 2022年第3期4卷 116-123页

作者： RenéBergmann Giuseppe Gulotta Federica Andreoni Tomoko Sumitomo Shigetada Kawabata Annelies S.Zinkernagel Gursharan S.Chhatwal Victor Nizet Manfred Rohde Satoshi Uchiyama Central Unit for Microscopy(ZEIM) Helmholtz Centre for Infection ResearchBraunschweigGermany Division of Infectious Diseases and Hospital Epidemiology University Hospital ZurichZurichSwitzerland Department of Oral and Molecular Microbiology Osaka University Graduate School of DentistrySuita-OsakaJapan Division of Host-Microbe Systems and Therapeutics Department of PediatricsUniversity of CaliforniaSan DiegoLa JollaCalifornia

Autophagy serves an innate immune function in defending the host against invading bacteria,including group a streptococcus(GAS).Autophagy is regulated by numerous host proteins,including the endogenous negative regulator calpain,a cytosolic protease.Globally disseminated serotypeM1T1 GAS strains associated with high invasive disease potential express numerous virulence factors and resist autophagic clearance.Upon in vitro infection of human epithelial cell lines with representative wild-type GAS M1T1 strain 5448(M1.5448),we observed increased calpain activation linked to a specific GAS virulence factor,the interleukin-8 protease SpyCEP.Calpain activation inhibited autophagy and decreased capture of cytosolic GAS in autophagosomes.In contrast,the serotype M6 GAS strain JRS4(M6.JRS4),which is highly susceptible to host autophagy-mediated killing,expresses low levels of SpyCEP and does not activate calpain.Overexpression of SpyCEP in M6.JRS4 stimulated calpain activation,inhibited autophagy,and significantly decreased bacterial capture in autophagosomes.These paired loss-and gain-of-function studies reveal a novel role for the bacterial protease SpyCEP in enabling GAS M1 evasion of autophagy and host innate immune clearance.

关键词： group a streptococcus SpyCEP virulence factor epithelial cells intracellular survival autophagy xenophagy calpain

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A MyD88-JAK1-STAT1 complex directly induces SOCS-1 expression in macrophages infected with group a streptococcus

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Cellular & Molecular Immunology 2015年第3期12卷 373-383页

作者： Jinghua Wu Cuiqing Ma Haixin Wang Shuhui Wu Gao Xue Xinli Shi Zhang Song Lin Wei Hebei Medical University Shijiazhuang China Hebei United University Affiliated Hospital Tangshan China Tangshan City Hospital of TraditionalChinese Medicine Tangshan China

Some pathogens can use host suppressor of cytokine signaling I （SOCS-1）, an important negative-feedback molecule, as the main mode of immune evasion. Here we found that group a streptococcus （GAS） is capable of inducing SOCS-1 expression in RAW264.7 and BMDM macrophages. IFN-p plays a role in GAS-induced SOCS-1 expression in macrophages following the induction of cytokine expression by GAS, representing the classical pathway of SOCS-1 expression. However, GAS also induced STAT1 activation and SOCS-1 expression when GAS-infected cells were incubated with anti-IFN-p monoclonal antibody in this study. Moreover, upon comparing TLR4-/- BMDM macrophages with wild-type （WT） cells, we found that TLR4 also plays an essential role in the induction of SOCS-1. MyD88, which is an adaptor protein for TLR4, contributes to STAT1 activation and phosphorylation by forming a complex with Janus kinase 1 （JAK1） and signal transducer and activator of transcription 1 （STAT1） in macrophages. GAS-stimulated expression of STAT1 was severely impaired in MyD88-/- macrophages, whereas expression of JAK1 was unaffected, suggesting that MyD88 was involved in STAT1 expression and phosphorylation. Together, these data demonstrated that in addition to IFN-p signaling and MyD88 complex formation, JAK1 and STAT1 act in a novel pathway to directly induce SOCS-1 expression in GAS-infected macrophages, which may be more conducive to rapid bacterial infection.

关键词： group a streptococcus macrophages SOCS-1 TLR4/MyD88 signaling

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Cellular & Molecular Immunology 2009年第1期6卷 73-77页

作者： Cuiqing Ma Caihong Li Xiurong Wang Ruihong Zeng Xiaolin Yin Huidong Feng Lin Wei Department of Immunology Basic Medical College Hebei Medical University Shijiazhuang 050017 China

group a streptococcus （GAS）, an important human pathogen, can cause various kinds of infections including superficial infections and potentially lethal infections, and the search for an effective vaccine to prevent GAS infections has been ongoing for many years. This paper compares the immunogenicity and immunoprotection of FbaA （an Fn-binding protein expressed on the surface of GAS） with that of M protein, the best immunogen of GAS. Assay for immune response showed that FbaA, similar to M protein, could induce protein-specific high IgG titer in BALB/c mice. Furthermore, following GAS challenge, the mice immunized with FbaA showed the same protective rate as those with M protein. These results indicate that FbaA is similar in ability to M protein in inducing protective immunity against GAS challenge in mice. Cellular ＆ Molecular Immunology.

关键词： group a streptococcus immunogenicity FbaA M protein immunoprotection

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group A streptococcal antigen exposed rat model to investigate neurobehavioral and cardiac complications associated with post-streptococcal autoimmune sequelae

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Animal Models and Experimental Medicine 2021年第2期4卷 151-161页

作者： Rukshan A.M.Rafeek Catherine M.Lobbe Ethan C.Wilkinson Adam S.Hamlin Nicholas M.Andronicos David J.McMillan Kadaba S.Sriprakash Natkunam Ketheesan School of Science&Technology University of New EnglandArmidaleNSWAustralia School of Science TechnologyEngineering and Genecology Research CentreUniversity of the Sunshine CoastMaroochydore DCQLDAustralia QIMR Berghofer Medical Research Institute HerstonQLDAustralia

Background:The neuropsychiatric disorders due to post-streptococcal autoimmune complications such as Sydenham's chorea(SC)are associated with acute rheumatic fever and rheumatic heart disease(ARF/RHD).An animal model that exhibits char-acteristics of both cardiac and neurobehavioral defects in ARF/RHD would be an important adjunct for future studies.Since age,gender,strain differences,and geno-types impact on the development of autoimmunity,we investigated the behavior of male and female Wistar and Lewis rat strains in two age cohorts(6 weeks and 12 weeks)under normal husbandry conditions and following exposure to group a streptococcus(GAS).Methods:Standard behavioral assessments were performed to determine the impair-ments in fine motor control(food manipulation test),gait and balance(beam walk-ing test),and obsessive-compulsive behavior(grooming and marble burying tests).Furthermore,electrocardiography,histology,and behavioral assessments were per-formed on male and female Lewis rats injected with GAS antigens.Results:For control Lewis rats there were no significant age and gender dependent differences in marble burying,food manipulation,beam walking and grooming be-haviors.In contrast significant age-dependent differences were observed in Wistar rats in all the behavioral tests except for food manipulation.Therefore,Lewis rats were selected for further experiments to determine the effect of GAS.After ex-posure to GAS,Lewis rats demonstrated neurobehavioral abnormalities and cardiac pathology akin to SC and ARF/RHD,respectively.Conclusion:We have characterised a new model that provides longitudinal stability of age-dependent behavior,to simultaneously investigate both neurobehavioral and cardiac abnormalities associated with post-streptococcal complications.

关键词： animal model group a streptococcus Lewis rats paediatric autoimmune neuropsychiatric disorders associated with streptococcal infections(PANDAS) rheumatic fever rheumatic heart disease sydenham chorea(SC)

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Site-Specific Conjugation of Cell Wall Polyrhamnose to Protein SpyAD Envisioning a Safe Universal group A Streptococcal Vaccine

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Infectious Microbes & Diseases 2021年第2期3卷 87-100页

作者： Nina J.Gao Satoshi Uchiyama Lucy Pill Samira Dahesh Joshua Olson Leslie Bautista Shilpa Maroju Aym Berges Janet ZLiu Raymond HZurich Nina Mvan Sorge Jeff Fairman Neeraj Kapoor Victor Nizet Division of Host-Microbe Systems and Therapeutics Department of PediatricsUC San DiegoLa JollaCaliforniaUSA Vaxcyte Inc.Foster CityCaliforniaUSA Department of Medical Microbiology and Infection Prevention Amsterdam University Medical CenterUniversity of AmsterdamAmsterdamThe Netherlands Netherlands Reference Laboratory for Bacterial Meningitis Amsterdam University Medical CenterUniversity of AmsterdamAmsterdamThe Netherlands Skaggs School of Pharmacy and Pharmaceutical Sciences UC San DiegoLa JollaCaliforniaUSA

Development of an effective vaccine against the leading human bacterial pathogen group a streptococcus(GAS)is a public health priority.The species defining group A cell wall carbohydrate(GAC,Lancefield antigen)can be engineered to remove its immunodominant N-acetylglucosamine(GlcNAc)side chain,implicated in provoking autoimmune cross-reactivity in rheumatic heart disease,leaving its polyrhamnose core(GACPR).Here we generate a novel protein conjugate of the GACPR and test the utility of this conjugate antigen in active immunization.Instead of conjugation to a standard carrier protein,we selected SpyAD,a highly conserved GAS surface protein containing both B-cell and T-cell epitopes relevant to the bacterium that itself shows promise as a vaccine antigen.SpyAD was synthesized using the XpressTM cell-free protein expression system,incorporating a non-natural amino acid to which GACpr was conjugated by site-specific click chemistry to yield high molecular mass SpyAD-GACPR conjugates and avoid disruption of important T-cell and B-cell immunological epitopes.The conjugated SpyAD-GACPR elicited antibodies that bound the surface of multiple GAS strains of diverse M types and promoted opsonophagocytic killing by human neutrophils.Active immunization of mice with a multivalent vaccine consisting of SpyAD-GACPR,together with candidate vaccine antigens streptolysin O and C5a peptidase,protected against GAS challenge in a systemic infection model and localized skin infection model,without evidence of cross reactivity to human heart or brain tissue epitopes.This general approach may allow GAC to be safely and effectively included in future GAS subunit vaccine formulations with the goal of broad protection without autoreactivity.

关键词： group a streptococcus streptococcus pyogenes group A carbohydrate Lancefield antigen conjugate vaccine XpressCFTM non-natural amino acids SpyAD Introduction

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重视A族链球菌和相关儿科疾病的研究

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临床儿科杂志 2006年第6期24卷 443-444页

作者：杨永弘沈叙庄首都医科大学附属北京儿童医院北京100045

A族链球菌(GAS)是儿童期感染的常见病原菌,可以造成严重侵袭性感染及感染后并发症。近年来,A族链球菌感染暴发报道增多,已引起人们关注,对A族链球菌基础和临床研究集中在感染的实验室快速诊断技术、临床菌株分型、流行病学、严重感染及... 详细信息

A族链球菌(GAS)是儿童期感染的常见病原菌,可以造成严重侵袭性感染及感染后并发症。近年来,A族链球菌感染暴发报道增多,已引起人们关注,对A族链球菌基础和临床研究集中在感染的实验室快速诊断技术、临床菌株分型、流行病学、严重感染及并发症发病机理等诸多方面。国内有关GAS研究报道尚不多,现将就其国外新近研究进展做一介绍。

关键词： group a streptococcus infection children

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Analysis of 11 cases of neonatal invasive streptococcus pyogenes infection

Analysis of 11 cases of neonatal invasive Streptococcus pyog...

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2019年浙江省医学会医学微生物与免疫学分会学术大会

作者： Yong-Ping Xie Chun-Zhen Hua Hui Yu Hong-Mei Xu Wei Gao Zhi-Wei Xu Ai-Wei Lin Division of Infectious Diseases The Children's HospitalZhejiang University School of Medicine Division of Infectious Diseases Children's Hospital of Fudan University Division of Infectious Diseases Children's Hospital of Chongqing Medical College Division of Infectious Diseases Kaifeng Children's Hospital Division of Infectious Diseases The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medicial University Division of Infectious Diseases Qilu Children's Hospital of Shandong University

Objective:To improve the understanding of clinical characteristics of neonatal iSPI.Methods:A retrospective study was performed to analyze the clinical and laboratory aspects of culture-confirmed neonatal iSPI cases during2010-2017.Results:Eleven cases of neonatal iSPI were confirmed.The mean age was 11.5±9.2 days.Ten cases had positive blood culture results and 1 had positive cerebrospinal fluid culture for streptococcus pyogenes.Seven patients had fever and 7 had identified focal infection,including 5 skin and soft tissue infections,1 case of pneumonia,and 1 skin burn with infection.All of the streptococcus pyogenes strains were sensitive to penicillin,cefotaxime,and vancomycin,and 81.8% were resistant to clindamycin and erythromycin.Six cases received penicillin or cephalosporin monotherapy.Three cases received meropenem plus vancomycin initially,and then two of them were switched to penicillin and one to vancomycin after pathogen identification.Two cases were treated with vancomycin monotherapy.All cases were discharged after being cured or showing improvement.Conclusion:Skin soft tissue infection is a common cause of neonatal iSPI.streptococcus pyogenes strains are highly sensitive to β-lactams,which should be used as the first-choice treatment.

关键词： streptococcus pyogenes group a streptococcus Neonate Invasive Infection

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Novel FbaA- and M protein-based multi-epitope vaccine elicits effective immune protection against group A streptococcui

Novel FbaA- and M protein-based multi-epitope vaccine elicit...

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第八届全国免疫学学术大会

作者： Cuiqing Ma zheng Liu Lin Wei Department of Immunology Basic Medical College Hebei Medical University

A recombinant multi-epitope vaccine of group a streptococcus,containing 7 predominant epitopes of FbaA identified by phages display technology and 5 non-tissue-cross-reactive M protein fragments from 4 different serotypes prevalent in China according to the bioinformatics analysis, was constructed by recombinant tech-niques.On the N-terminal of the vaccine we designed Trojan peptide sequence for the promotion of antigen presenting cells and Poly Alamine DR Epitope for balance of Th lymphocyte responses.BALB/c mice were immunized subcutaneously with the multivalent vaccine formulated with Freund adjuvant to contain 50μg protein per dose, and recombinant FbaA and M protein were performed in parallel as control.Immune sera were assayed for the Ig G titres, the Ig G subclasses and its affinity with M1 GAS strain by enzyme-linked immunosorbent assay, Results indicated that the multivalent vaccine had highly immunogenic and elicited a balanced IgG 1/IgG 2a(Th2/Th1) response.Interestingly, the positive rate, when the multivalent vaccine acted as a antigen to detect ASO-positive patient sera, was 95.8%, indicating that the epitopes of the vaccine were expressed widely in the prevalent serotypes of group a streptococcus.Importantly, the vaccine triggered a robust protective response(survival rate 80%), meanwhile, none of the immune sera cross-reacting with human tissues.

关键词： group a streptococcus Vaccine immune protection Fba protein M protein

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