Milad Parvane, S. Nazarian, Emad Kordbache, Javad Fathi, Mohamad Ebrahim Minae, Mohammad Reza Ramezani
{"title":"plga包封重组GroEL鼠伤寒沙门氏菌对肠出血性和肠致病性大肠杆菌免疫应答的评价","authors":"Milad Parvane, S. Nazarian, Emad Kordbache, Javad Fathi, Mohamad Ebrahim Minae, Mohammad Reza Ramezani","doi":"10.18502/ajmb.v14i4.10484","DOIUrl":null,"url":null,"abstract":"Background: Heat Shock Proteins (HSPs) elicit humoral and cellular immune responses. Due to their high sequence homology, they can be developed as a new immunogen for cross prophylactic and vaccination effects against infectious agents such as Enteropathogenic and Enterohemorrhagic Escherichia coli (EPEC and EHEC). This study aimed to evaluate the immunogenicity and cross-protective efficacy of rGroEL of Salmonella typhi (S. typhi) encapsulated in poly lactic-co-glycolic acid (PLGA) nanoparticles against EPEC and EHEC. Methods: Recombinant GroEL was expressed in Escherichia coli (E. coli) and purified using Ni-NTA affinity chromatography. The protein was encapsulated in PLGA by the double emulsion method, and the nanoparticles were characterized physicochemically. BALB/c mice were immunized, and the efficacy of the protein to elicit immune responses was assessed. Results: Over-expression in E. coli led to corresponding 64.5 kDa protein bands in Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Non-aggregated nanoparticles had a spherical shape with a mean diameter of 194.3±3 nm and encapsulation efficiency of 89.5±2.5%. Antibody isotyping revealed that GroEL immunization induced both IgG1 and IgG2a antibodies. Moreover, immunization of the mice with recombinant GroEL protein conferred 80 and 60% protection against lethal infections by EPEC and EHEC, respectively. Furthermore, organ burden studies revealed a significant reduction in infection in the immunized mice compared to the non-immunized ones. Passive immunization with anti-GroEL sera also protected 50% of the mice against the lethal doses of EHEC and EPEC strains. Conclusion: The findings indicated that immunization of the mice with recombinant GroEL of S. typhi elicited cross-protection against other bacterial infections. This represented the immense potential of GroEL to be developed as a single vaccine against multiple pathogens.","PeriodicalId":8669,"journal":{"name":"Avicenna journal of medical biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2022-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":"{\"title\":\"Evaluation of PLGA-Encapsulated Recombinant GroEL of S. typhi immune Responses Against Enterohaemorrhagic and Enteropathogenic Escherichia coli\",\"authors\":\"Milad Parvane, S. Nazarian, Emad Kordbache, Javad Fathi, Mohamad Ebrahim Minae, Mohammad Reza Ramezani\",\"doi\":\"10.18502/ajmb.v14i4.10484\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Background: Heat Shock Proteins (HSPs) elicit humoral and cellular immune responses. Due to their high sequence homology, they can be developed as a new immunogen for cross prophylactic and vaccination effects against infectious agents such as Enteropathogenic and Enterohemorrhagic Escherichia coli (EPEC and EHEC). This study aimed to evaluate the immunogenicity and cross-protective efficacy of rGroEL of Salmonella typhi (S. typhi) encapsulated in poly lactic-co-glycolic acid (PLGA) nanoparticles against EPEC and EHEC. Methods: Recombinant GroEL was expressed in Escherichia coli (E. coli) and purified using Ni-NTA affinity chromatography. The protein was encapsulated in PLGA by the double emulsion method, and the nanoparticles were characterized physicochemically. BALB/c mice were immunized, and the efficacy of the protein to elicit immune responses was assessed. Results: Over-expression in E. coli led to corresponding 64.5 kDa protein bands in Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Non-aggregated nanoparticles had a spherical shape with a mean diameter of 194.3±3 nm and encapsulation efficiency of 89.5±2.5%. Antibody isotyping revealed that GroEL immunization induced both IgG1 and IgG2a antibodies. Moreover, immunization of the mice with recombinant GroEL protein conferred 80 and 60% protection against lethal infections by EPEC and EHEC, respectively. Furthermore, organ burden studies revealed a significant reduction in infection in the immunized mice compared to the non-immunized ones. Passive immunization with anti-GroEL sera also protected 50% of the mice against the lethal doses of EHEC and EPEC strains. Conclusion: The findings indicated that immunization of the mice with recombinant GroEL of S. typhi elicited cross-protection against other bacterial infections. This represented the immense potential of GroEL to be developed as a single vaccine against multiple pathogens.\",\"PeriodicalId\":8669,\"journal\":{\"name\":\"Avicenna journal of medical biotechnology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-09-03\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Avicenna journal of medical biotechnology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.18502/ajmb.v14i4.10484\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Avicenna journal of medical biotechnology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.18502/ajmb.v14i4.10484","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
Evaluation of PLGA-Encapsulated Recombinant GroEL of S. typhi immune Responses Against Enterohaemorrhagic and Enteropathogenic Escherichia coli
Background: Heat Shock Proteins (HSPs) elicit humoral and cellular immune responses. Due to their high sequence homology, they can be developed as a new immunogen for cross prophylactic and vaccination effects against infectious agents such as Enteropathogenic and Enterohemorrhagic Escherichia coli (EPEC and EHEC). This study aimed to evaluate the immunogenicity and cross-protective efficacy of rGroEL of Salmonella typhi (S. typhi) encapsulated in poly lactic-co-glycolic acid (PLGA) nanoparticles against EPEC and EHEC. Methods: Recombinant GroEL was expressed in Escherichia coli (E. coli) and purified using Ni-NTA affinity chromatography. The protein was encapsulated in PLGA by the double emulsion method, and the nanoparticles were characterized physicochemically. BALB/c mice were immunized, and the efficacy of the protein to elicit immune responses was assessed. Results: Over-expression in E. coli led to corresponding 64.5 kDa protein bands in Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Non-aggregated nanoparticles had a spherical shape with a mean diameter of 194.3±3 nm and encapsulation efficiency of 89.5±2.5%. Antibody isotyping revealed that GroEL immunization induced both IgG1 and IgG2a antibodies. Moreover, immunization of the mice with recombinant GroEL protein conferred 80 and 60% protection against lethal infections by EPEC and EHEC, respectively. Furthermore, organ burden studies revealed a significant reduction in infection in the immunized mice compared to the non-immunized ones. Passive immunization with anti-GroEL sera also protected 50% of the mice against the lethal doses of EHEC and EPEC strains. Conclusion: The findings indicated that immunization of the mice with recombinant GroEL of S. typhi elicited cross-protection against other bacterial infections. This represented the immense potential of GroEL to be developed as a single vaccine against multiple pathogens.