Immunization with Bivalent Flagellin Protects Mice against Fatal Pseudomonas aeruginosa Pneumonia

Pseudomonas aeruginosa lung infections present a major challenge to healthcare systems worldwide because they are commonly associated with high morbidity and mortality. Here, we demonstrate the protective efficacy of type a and b flagellins (bivalent flagellin) against acute fatal pneumonia in mice. Mice immunized intranasally with a bivalent flagellin vaccine were challenged by different flagellated strains of P. aeruginosa in an acute pneumonia model. Besides the protective effect of the vaccine, we further measured the host innate and cellular immunity responses. The immunized mice in our study were protected against both strains. Remarkably, active immunization with type a or b flagellin significantly improved survival of mice against heterologous strain compared to flagellin a or b antisera. We also showed that after an intranasal challenge by P. aeruginosa strain, neutrophils are recruited to the airways of vaccinated mice, and that the bivalent flagellin vaccine was proved to be protective by the generated CD4+IL-17+ Th17 cells. In conclusion, bivalent flagellin vaccine can confer protection against different strains of P. aeruginosa in an acute pneumonia mouse model by eliciting effective cellular and humoral immune responses, including increased IL-17 production and improved opsonophagocytic killing

Wide distribution of carbapenem resistant Acinetobacter baumannii in burns patients in Iran

Antimicrobial resistance in carbapenem non-susceptible Acinetobacter baumannii (CNSAb) is a major public health concern globally. This study determined the antibiotic resistance and molecular epidemiology of CNSAb isolates from a referral burn center in Tehran, Iran. Sixty-nine CNSAb isolates were tested for susceptibility to antimicrobial agents using the E test methodology. Multiple locus variable number tandem repeat analysis (MLVA), Multilocus sequence typing (MLST) and multiplex PCR were performed. PCR assays tested for ambler classes A, B, and D β-lactamases. Detection of ISAba1, characterization of integrons, and biofilm formation were investigated. Fifty-three (77) isolates revealed XDR phenotypes. High prevalence of blaOXA-23-like (88) and blaPER-1 (54) were detected. ISAba1 was detected upstream of blaADC, blaOXA-23-like and blaOXA51-like genes in, 97, 42, and 26 of isolates, respectively. Thirty-one (45) isolates were assigned to international clone (IC) variants. MLVA identified 56 distinct types with six clusters and 53 singleton genotypes. Forty previously known MLST sequence types forming 5 clonal complexes were identified. The Class 1 integron (class 1 integrons) gene was identified in 84 of the isolates. The most prevalent (33) cassette combination was aacA4-catB8-aadA1. The IC variants were predominant in the A. baumannii lineage with the ability to form strong biofilms. The XDR-CNSAb from burned patients in Iran is resistant to various antimicrobials, including tigecycline. This study shows wide genetic diversity in CNSAb. Integrating the new Iranian A. baumannii IC variants into the epidemiologic clonal and susceptibility profile databases can help effective global control measures against the XDR-CNSAb pandemic. � 2015 Farshadzadeh, Hashemi, Rahimi, Pourakbari, Esmaeili, Haghighi, Majidpour, Shojaa, Rahmani, Gharesi, Aziemzadeh and Bahador

Wide distribution of carbapenem resistant Acinetobacter baumannii in burns patients in Iran

Antimicrobial resistance in carbapenem non-susceptible Acinetobacter baumannii (CNSAb) is a major public health concern globally. This study determined the antibiotic resistance and molecular epidemiology of CNSAb isolates from a referral burn center in Tehran, Iran. Sixty-nine CNSAb isolates were tested for susceptibility to antimicrobial agents using the E test methodology. Multiple locus variable number tandem repeat analysis (MLVA), Multilocus sequence typing (MLST) and multiplex PCR were performed. PCR assays tested for ambler classes A, B, and D β-lactamases. Detection of ISAba1, characterization of integrons, and biofilm formation were investigated. Fifty-three (77) isolates revealed XDR phenotypes. High prevalence of blaOXA-23-like (88) and blaPER-1 (54) were detected. ISAba1 was detected upstream of blaADC, blaOXA-23-like and blaOXA51-like genes in, 97, 42, and 26 of isolates, respectively. Thirty-one (45) isolates were assigned to international clone (IC) variants. MLVA identified 56 distinct types with six clusters and 53 singleton genotypes. Forty previously known MLST sequence types forming 5 clonal complexes were identified. The Class 1 integron (class 1 integrons) gene was identified in 84 of the isolates. The most prevalent (33) cassette combination was aacA4-catB8-aadA1. The IC variants were predominant in the A. baumannii lineage with the ability to form strong biofilms. The XDR-CNSAb from burned patients in Iran is resistant to various antimicrobials, including tigecycline. This study shows wide genetic diversity in CNSAb. Integrating the new Iranian A. baumannii IC variants into the epidemiologic clonal and susceptibility profile databases can help effective global control measures against the XDR-CNSAb pandemic. � 2015 Farshadzadeh, Hashemi, Rahimi, Pourakbari, Esmaeili, Haghighi, Majidpour, Shojaa, Rahmani, Gharesi, Aziemzadeh and Bahador

Immunogenicity and protective efficacy of Pseudomonas aeruginosa type a and b flagellin vaccines in a burned mouse model

Immunogenicity and efficacy of Pseudomonas aeruginosa type a and b flagellins (hereafter, flagellins) as candidate vaccines were evaluated using an experimental burned mouse model. The protection afforded and the reduction in bacterial burden achieved by these vaccine candidates were determined. Primary immunization with flagellins followed by two booster shots generated a robust immune response. Cytokine analysis demonstrated the secretion of interleukin-4 more than interferon-γ from immunized T-cells in response to in vitro antigen stimulation. IgG response was of Th2 type, predominantly with IgG1 and lower IgG2a levels before and after challenge. In vitro opsonophagocytosis assays confirmed protective potential of immune sera via enhanced bacterial cell killing. Immune sera also inhibited P. aeruginosa motility. Serum cytokine analysis demonstrated high IL-12 and low IL-10 levels in flagellin-immunized mouse sera. Reduced systemic bacterial spread from original infection site into liver and spleen was associated with increased survival. Immunization of mice with flagellins increased the humoral immune response and protection against P. aeruginosa infection in our mouse model. © 2016 Elsevier Ltd

Flagellin and pilin immunization against multi-drug resistant Pseudomonas aeruginosa protects mice in the burn wound sepsis model

Pseudomonas aeruginosa is a formidable pathogen and a major threat to burn patients. Antimicrobial therapy is often unsuccessful because P. aeruginosa can develop multi-drug resistance; thus, immunotherapy and vaccine can be a rational alternative. Flagella and type IV pili have been identified as important virulence factors in the colonization and pathogenesis of P. aeruginosa in burn wound infections. Immunogenicity and efficacy of mixed recombinant full-length type b flagellin (r-b-flagellin) and recombinant PilA (r-PilA) as candidate vaccines were assessed by measuring humoral and cellular responses, using an experimental burned mouse model. Primary immunization with "r-b-flagellin + r-PilA" followed by two booster shots was sufficient to generate a robust humoral response, which was predominantly a Th2 response consisting mainly of subtype IgG1 and low levels of IgG2a. Analysis of the cytokine response among immunized mice showed an increased production of IL-4, INF-γ and IL-17 by splenocytes upon stimulation by "r-b-flagellin + r-PilA". Opsonophagocytosis assays confirmed the enhanced killing of bacteria by anti "r-b-flagellin + r-PilA" immune sera. These antibodies were also able to reduce bacterial load in the site of original infection into the liver and spleen of challenged mice. The reduction of systemic bacterial spread resulted in an increased survival rate of challenged immunized mice. In conclusion, immunization with "r-b-flagellin + r-PilA" proteins provides a better protective response against P. aeruginosa infection in the burn mouse model. © 2016 European Federation of Immunological Societies

Immunogenicity of Pseudomonas aeruginosa recombinant b-type fagellin as a vaccine candidate: Protective efficacy in a murine burn wound sepsis model

Pseudomonas aeruginosa (PA) is a formidable opportunistic pathogen among patients with burn wound infections. Antimicrobial therapy is often unsuccessful because PA can develop multi-drug resistance; thus, immunotherapy can be a rational alternative. The goal of this study was to evaluate the immunogenicity recombinant type b flagellin (r-b-flagellin) as a potential vaccine against P. aeruginosa in a mouse model for burn wound sepsis. Primary immunization with r-b-flagellin (10. μg) followed by two booster shots was sufficient to generate a robust humoral response, which was predominantly a T helper 2 (Th2) type response consisting mainly of subtype IgG1 and low levels of IgG2a. Analysis of the Th1-Th2 response among immunized mice showed an increased production of IL-4, INF-γ and IL-17 by splenocytes upon stimulation by r-b-flagellin. Opsono-phagocytosis assays confirmed the enhanced killing of bacteria by anti r-b-flagellin immune sera. These antibodies were also able to inhibit motility of P. aeruginosa and afforded protection to immunized mice by reducing bacterial load in the site of original infection into the liver of challenged mice. The reduction of systemic bacterial spread resulted in an increase in the survival rate of challenged immunized mice. In conclusion, immunization of mice with r-b-flagellin protein increased the level of humoral and cellular immune response and led to an efficacious protection against P. aeruginosa infection in the burn mouse model. © 2016 Elsevier Ltd and ISBI

Protective effect of pilin protein with alum+naloxone adjuvant against acute pulmonary Pseudomonas aeruginosa infection

Pseudomonas aeruginosa is an important opportunistic human pathogen that causes a wide variety of severe nosocomial infections. Type IV pili of P. aeruginosa are made up of polymerized pilin that aids in bacterial adhesion, biofilm formation and twitching motility. The aim of this study was to evaluate the efficacy of alum and naloxone (alum+NLX) as an adjuvant for P. aeruginosa recombinant PilA (r-PilA) as a vaccine candidate in the improvement of humoral and cellular immunity. Primary immunization with r-PilA in combination with alum+NLX followed by two booster shots was sufficient to generate robust cellular and humoral responses, which were Th1 and Th2 type responses consisting of IgG1 and IgG2a subtypes. Analysis of the cytokine response among immunized mice showed an increased production of IL-4, INF-γ and IL-17 by splenocytes upon stimulation by r-PilA. These sera were also able to reduce bacterial load in the lung tissue of challenged mice. The reduction of systemic bacterial spread resulted in increased survival rates in challenged immunized mice. In conclusion, immunization with r-PilA combined with alum+NLX evokes cellular and humoral immune responses, which play an important role in providing protection against acute P. aeruginosa lung infection among immunized mice. © 2016 International Alliance for Biological Standardizatio