scholarly journals Enhanced Immunogenicity in the Murine Airway Mucosa with an Attenuated Salmonella Live Vaccine Expressing OprF-OprI from Pseudomonas aeruginosa

2004 ◽  
Vol 72 (11) ◽  
pp. 6546-6553 ◽  
Author(s):  
Heinz Arnold ◽  
Dirk Bumann ◽  
Melanie Felies ◽  
Britta Gewecke ◽  
Meike Sörensen ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Outer Membrane Proteins ◽  
Immunoglobulin A ◽  
Vaccination Strategy ◽  
Booster Vaccination ◽  
Primary Vaccination ◽  
Live Vaccine ◽  
Igg Antibody ◽  
Inducible Protein

ABSTRACT We constructed an oral live vaccine based on the attenuated aroA mutant Salmonella enterica serovar Typhimurium strain SL3261 expressing outer membrane proteins F and I (OprF-OprI) from Pseudomonas aeruginosa and investigated it in a mouse model. Strains with in vivo inducible protein expression with the P pacC promoter showed good infection rates and immunogenicity but failed to engender detectable antibodies in the lung. However, a systemic booster vaccination following an oral primary immunization yielded high immunoglobulin A (IgA) and IgG antibody levels in both upper and lower airways superior to conventional systemic or mucosal booster vaccination alone. In addition, the proportion of IgG1 and IgG2a antibodies suggested that the systemic booster does not alter the more TH1-like type of response induced by the oral Salmonella primary vaccination. We conclude that an oral primary systemic booster vaccination strategy with an appropriate mucosal vector may be advantageous in diseases with the risk of P. aeruginosa airway infection, such as cystic fibrosis.

scholarly journals Role of B7 Costimulatory Molecules in Mediating Systemic and Mucosal Antibody Responses to Attenuated Salmonella enterica Serovar Typhimurium and Its Cloned Antigen

2004 ◽  
Vol 72 (10) ◽  
pp. 5824-5831 ◽  
Author(s):  
Carlos A. Garcia ◽  
Michael Martin ◽  
Suzanne M. Michalek
Keyword(s):  
Salmonella Enterica ◽  
Immunoglobulin A ◽  
Costimulatory Molecules ◽  
Antibody Responses ◽  
Wild Type ◽  
Igg Antibody ◽  
Functional Roles ◽  
Serovar Typhimurium

ABSTRACT The purpose of the present study was to evaluate the ability of an attenuated Salmonella enterica serovar Typhimurium vaccine strain to up-regulate B7-1 and B7-2 on antigen-presenting cells and to examine the functional roles these costimulatory molecules play in mediating immune responses to Salmonella and to an expressed cloned antigen, the saliva-binding region (SBR) of antigen I/II. In vitro stimulation of B cells (B220+), macrophages (CD11b+), and dendritic cells (CD11c+) with S. enterica serovar Typhimurium induced an up-regulation of B7-2 and, especially, B7-1 expression. The in vivo functional roles of B7-1, B7-2, and B7-1/2 were evaluated in BALB/c wild-type and B7-1, B7-2, and B7-1/2 knockout (KO) mice following intranasal immunization with the Salmonella expressing the cloned SBR. Differential requirements for B7-1 and B7-2 were observed upon primary and secondary immunizations. Compared to wild-type controls, B7-1 and B7-2 KO mice had reduced mucosal and systemic anti-Salmonella antibody responses after a single immunization, while only B7-1 KO mice exhibited suppressed anti-Salmonella antibody responses following the second immunization. Mucosal and systemic antibody responses to SBR were reduced following the primary immunization, whereas a compensatory role for either B7-1 or B7-2 was observed after the second immunization. B7-1/2 double KO mice failed to induce detectable levels of mucosal or systemic immunoglobulin A (IgA) or IgG antibody responses to either Salmonella or SBR. These findings demonstrate that B7-1 and B7-2 can play distinct as well as redundant roles for mediating mucosal and systemic antibody responses, which are likely dependent upon the nature of the antigen.

scholarly journals Salmonella chitosan nanoparticle vaccine administration is protective against Salmonella Enteritidis in broiler birds

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259334
Author(s):  
Keila Y. Acevedo-Villanueva ◽  
Sankar Renu ◽  
Revathi Shanmugasundaram ◽  
Gabriel O. Akerele ◽  
Renukaradhy J. Gourapura ◽  
...  
Keyword(s):  
Salmonella Enteritidis ◽  
Body Weight Gain ◽  
Control Strategies ◽  
Outer Membrane Proteins ◽  
Booster Vaccination ◽  
Live Vaccine ◽  
Inos Mrna ◽  
Feed Conversion ◽  
T Lymphocyte Proliferation ◽  
Chitosan Nanoparticle

Salmonella control strategies include vaccines that help reduce the spread of Salmonella in poultry flocks. In this study we evaluated the efficacy of administering a live Salmonella vaccine followed by a killed Salmonella chitosan nanoparticle (CNP) vaccine booster on the cellular and humoral immunity of broilers. The CNP vaccine was synthesized with Salmonella Enteritidis (S. Enteritidis) outer-membrane-proteins (OMPs) and flagellin-proteins. At d1-of-age, one-hundred-sixty-eight chicks were allocated into treatments: 1) No vaccine, 2) Live vaccine (Poulvac®ST), 3) CNP vaccine, 4) Live+CNP vaccine. At d1-of-age, birds were orally vaccinated with PBS, Live vaccine, or CNP. At d7-of-age, the No vaccine, Live vaccine and CNP vaccine groups were boosted with PBS and the Live+CNP vaccine group was boosted with CNP. At d14-of-age, birds were challenged with 1×109 CFU/bird S. Enteritidis. There were no significant differences in body-weight-gain (BWG) or feed-conversion-ratio (FCR). At 8h-post-challenge, CNP and Live+CNP-vaccinated birds had 17% and 24% greater levels (P<0.05) of anti-Salmonella OMPs IgA in bile, respectively, compared to control. At d28-of-age, CNP, Live, and Live+CNP-vaccinated birds had 33%, 18%, and 24% greater levels (P<0.05) of anti-Salmonella OMPs IgA in bile, respectively, compared to control. At d14-of-age, Live+CNP-vaccinated birds had 46% greater levels (P<0.05) of anti-Salmonella OMPs IgY in serum, compared to control. At d21-of-age, splenocytes from CNP and Live-vaccinated birds had increased (P<0.05) T-lymphocyte proliferation at 0.02 mg/mL OMPs stimulation compared to the control. At d28-of-age, CNP and Live+CNP-vaccinated birds had 0.9 Log10 CFU/g and 1 Log10 CFU/g decreased S. Enteritidis cecal loads (P<0.05), respectively, compared to control. The CNP vaccine does not have adverse effects on bird’s BWG and FCR or IL-1β, IL-10, IFN-γ, or iNOS mRNA expression levels. It can be concluded that the CNP vaccine, as a first dose or as a booster vaccination, is an alternative vaccine candidate against S. Enteritidis in broilers.

Ethanol Extract Of Centella Asiatica (L.) Urban Leaves Effectively Inhibit Streptococcus pyogenes and Pseudomonas aeruginosa by Invitro Test

2020 ◽  
Vol 2 (2) ◽  
pp. 69-76
Author(s):  
Dini Aulia Azmi ◽  
Nurlailah Nurlailah ◽  
Ratih Dewi Dwiyanti
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Streptococcus Pyogenes ◽  
Bacterial Infections ◽  
Herbal Medicines ◽  
Centella Asiatica ◽  
Ethanol Extract ◽  
Dilution Method ◽  
Initial Stage ◽  
Independent Variable

Streptococcus pyogenes and Pseudomonas aeruginosa are some of the causes of infectious diseases. Centella asiatica (L.) Urban has many benefits for humans, including overcoming fever, anti-bacterial, and anti-inflammatory. This study aims to determine the inhibition of Centella asiatica (L.) Urban leaves ethanol extract on the growth of Streptococcus pyogenes and Pseudomonas aeruginosa. This research is the initial stage of the development of herbal medicines to treat Streptococcus pyogenes and Pseudomonas aeruginosa infections. The independent variable was the concentration of ethanol extract of Centella asiatica (L.) Urban leaves and the dependent variable was the growth of Streptococcus pyogenes and Pseudomonas aeruginosa. The anti-bacterial activity test was carried out by the liquid dilution method. The concentrations used are 20%, 40%, 60%, 80%. 100% The results showed that the minimum inhibitory concentration (MIC) against Streptococcus pyogenes: 40% and Pseudomonas aeruginosa: 40%. Minimum bactericidal concentration (MBC) results for Streptococcus pyogenes: 60% and Pseudomonas aeruginosa: 60%. So it can be concluded that there is inhibition of the ethanol extract of Centella asiatica (L.) Urban leaves on the growth of Streptococcus pyogenes and Pseudomonas aeruginosa. Centella Asiatica (L.) Urban extract has potential as herbal medicine against bacterial infections but requires further research to determine its effect in vivo.

Reduction of Milk Contamination with Aflatoxin-M1 through Vaccination of Dairy Cattle with Aflatoxin-B1 Vaccine

2020 ◽  
Keyword(s):  
Dairy Cattle ◽  
Aflatoxin B1 ◽  
Booster Vaccination ◽  
Primary Vaccination ◽  
Nano Particles ◽  
Aflatoxin M1 ◽  
Serum Samples ◽  
Serum Albumins ◽  
Milk Contamination ◽  
Before And After

Aflatoxin M1 is one of mycotoxin derivatives, which is secreted in milk of dairy cattle fed on feed contaminated with Aflatoxin-B1 (AFB1). The current study was designed to prepare a vaccine against AFB1and to evaluate its efficacy in reducing or preventing secretion of AFM1 in milk. Aflatoxin-B1 was prepared, purified and transformed into oxime, then it was fixed on bovine serum albumins. The AFB1-BSA conjugate was adjuvanted with Gold Nano particles then Montanide ISA 206. The prepared vaccine was used for immunization of rabbits by S/c routes as 100 µg/dose and dairy cattle by I/M routes as 500 µg/dose. The vaccinated animals were boosted at 3 weeks post primary immunization. Serum samples were collected and examined for the anti-AFB1 using AGPT. A mean titer of 15.2 AGPU/ml was detected at 2 weeks post primary vaccination then significantly increased till reached to 76.8 AGPU/ml at 6 weeks post Booster vaccination. All vaccinated rabbits were challenged with dose of 0.3 mg AFB1 toxin/Kg. The vaccinated rabbit showed 100% protection and no AFB1 toxin residue was detected in their livers. Milk samples were collected from non-vaccinated and AFB1-immunized dairy cattle then examined with ELISA for quantitation of AFM1 residues before and after vaccination. The results showed that the prepared AFB1 vaccine was safe, potent and able to reduce AFM1 release in milk of vaccinated heifers by 70%. So the vaccination of lactating animals with the AFB1vaccine might represent a valid tool for the prevention of AFM1 contamination of milk and dairy products.

scholarly journals Baicalin Represses Type Three Secretion System of Pseudomonas aeruginosa through PQS System

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1497
Author(s):  
Pansong Zhang ◽  
Qiao Guo ◽  
Zhihua Wei ◽  
Qin Yang ◽  
Zisheng Guo ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Mammalian Cells ◽  
Secretion System ◽  
Chinese Herbs ◽  
Infection Model ◽  
Lung Pathology ◽  
Promising Alternative ◽  
The Impact

Therapeutics that target the virulence of pathogens rather than their viability offer a promising alternative for treating infectious diseases and circumventing antibiotic resistance. In this study, we searched for anti-virulence compounds against Pseudomonas aeruginosa from Chinese herbs and investigated baicalin from Scutellariae radix as such an active anti-virulence compound. The effect of baicalin on a range of important virulence factors in P. aeruginosa was assessed using luxCDABE-based reporters and by phenotypical assays. The molecular mechanism of the virulence inhibition by baicalin was investigated using genetic approaches. The impact of baicalin on P. aeruginosa pathogenicity was evaluated by both in vitro assays and in vivo animal models. The results show that baicalin diminished a plenty of important virulence factors in P. aeruginosa, including the Type III secretion system (T3SS). Baicalin treatment reduced the cellular toxicity of P. aeruginosa on the mammalian cells and attenuated in vivo pathogenicity in a Drosophila melanogaster infection model. In a rat pulmonary infection model, baicalin significantly reduced the severity of lung pathology and accelerated lung bacterial clearance. The PqsR of the Pseudomonas quinolone signal (PQS) system was found to be required for baicalin’s impact on T3SS. These findings indicate that baicalin is a promising therapeutic candidate for treating P. aeruginosa infections.

scholarly journals Surrogate post-coital testing for contraceptive efficacy against human sperm activity in the ovine vaginal model†

2020 ◽  
Author(s):  
Yong Zhu ◽  
Jamal Saada ◽  
Shrestha Bhawana ◽  
Sam Lai ◽  
Paula Villarreal ◽  
...  
Keyword(s):  
Human Sperm ◽  
Vaginal Fluid ◽  
Human Semen ◽  
Ovine Model ◽  
Igg Antibody ◽  
Study Objective ◽  
In Vivo Testing ◽  
Efficacy Assessment ◽  
Sperm Immobilization

Abstract High unintended pregnancy rates are partially due to lack of effective nonhormonal contraceptives; development of safe, effective topical vaginal methods will address this need. Preclinical product safety and efficacy assessment requires in vivo testing in appropriate models. The sheep is a good model for the evaluation of vaginally delivered products due to its close similarities to humans. The study objective was to develop an ovine model for efficacy testing of female nonhormonal contraceptives that target human sperm. Fresh human semen was pooled from male volunteers. Nonpregnant female Merino sheep were treated with control or vaginal contraceptive product (IgG antibody with action against sperm or nonoxynol-9 [N9]). Pooled semen was added to the sheep vagina and mixed with product and vaginal secretions. Microscopic assessment of samples was performed immediately and progressive motility (PM) of sperm was compared between treatments. Cytokines CXCL8 and IL1B were assessed in vaginal fluid after instillation of human semen. No adverse reactions or elevations in proinflammatory cytokines occurred in response to human semen. N9 produced signs of acute cellular toxicity while there were no cellular changes after IgG treatment. N9 and IgG had dose-related effects with the highest dose achieving complete sperm immobilization (no sperm with PM). Surrogate post-coital testing of vaginally administered contraceptives that target human semen was developed in an ovine model established for vaginal product preclinical testing. This expanded model can aid the development of much needed nonhormonal topical vaginal contraceptives, providing opportunities for rapid iterative drug development prior to costly, time-intensive human testing.

scholarly journals Anti-Pseudomonas aeruginosa antibody detection in patients with bronchiectasis without cystic fibrosis

Thorax ◽  
2001 ◽  
Vol 56 (9) ◽  
pp. 669-674
Author(s):  
E Caballero ◽  
M-E Drobnic ◽  
M-T Pérez ◽  
J-M Manresa ◽  
A Ferrer ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Antibody Detection ◽  
Individual Bands ◽  
Diagnostic Usefulness ◽  
Group A ◽  
Group B

BACKGROUNDPseudomonas aeruginosa is a frequent cause of infection in patients with bronchiectasis. Differentiation between non-infected patients and those with different degrees of P aeruginosainfection could influence the management and prognosis of these patients. The diagnostic usefulness of serum IgG antibodies againstP aeruginosa outer membrane proteins was determined in patients with bronchiectasis without cystic fibrosis.METHODSFifty six patients were classified according to sputum culture into three groups: group A (n=18) with no P aeruginosain any sample; group B (n=18) with P aeruginosa alternating with other microorganisms; and group C (n=20) with P aeruginosa in all sputum samples. Each patient had at least three sputum cultures in the 6 months prior to serum collection. Detection of antibodies was performed by Western blot and their presence against 20 protein bands (10–121 kd) was assessed.RESULTSAntibodies to more than four bands in total or to five individual bands (36, 26, 22, 20 or 18 kd) differentiated group B from group A, while antibodies to a total of more than eight bands or to 10 individual bands (104, 69, 63, 56, 50, 44, 30, 25, 22, 13 kd) differentiated group C from group B. When discordant results between the total number of bands and the frequency of P aeruginosa isolation were obtained, the follow up of patients suggested that the former, in most cases, predicted chronic P aeruginosacolonisation.CONCLUSIONIn patients with bronchiectasis the degree of P aeruginosa infection can be determined by the number and type of outer membrane protein bands indicating which serum antibodies are present.

scholarly journals A Novel Infection Protocol in Zebrafish Embryo to Assess Pseudomonas aeruginosa Virulence and Validate Efficacy of a Quorum Sensing Inhibitor In Vivo

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 401
Author(s):  
Pauline Nogaret ◽  
Fatima El El Garah ◽  
Anne-Béatrice Blanc-Potard
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Animal Model ◽  
Quorum Sensing ◽  
Drug Testing ◽  
Drug Efficacy ◽  
Embryo Viability ◽  
Immersion Method ◽  
Opportunistic Human Pathogen

The opportunistic human pathogen Pseudomonas aeruginosa is responsible for a variety of acute infections and is a major cause of mortality in chronically infected cystic fibrosis patients. Due to increased resistance to antibiotics, new therapeutic strategies against P. aeruginosa are urgently needed. In this context, we aimed to develop a simple vertebrate animal model to rapidly assess in vivo drug efficacy against P. aeruginosa. Zebrafish are increasingly considered for modeling human infections caused by bacterial pathogens, which are commonly microinjected in embryos. In the present study, we established a novel protocol for zebrafish infection by P. aeruginosa based on bath immersion in 96-well plates of tail-injured embryos. The immersion method, followed by a 48-hour survey of embryo viability, was first validated to assess the virulence of P. aeruginosa wild-type PAO1 and a known attenuated mutant. We then validated its relevance for antipseudomonal drug testing by first using a clinically used antibiotic, ciprofloxacin. Secondly, we used a novel quorum sensing (QS) inhibitory molecule, N-(2-pyrimidyl)butanamide (C11), the activity of which had been validated in vitro but not previously tested in any animal model. A significant protective effect of C11 was observed on infected embryos, supporting the ability of C11 to attenuate in vivo P. aeruginosa pathogenicity. In conclusion, we present here a new and reliable method to compare the virulence of P. aeruginosa strains in vivo and to rapidly assess the efficacy of clinically relevant drugs against P. aeruginosa, including new antivirulence compounds.

scholarly journals DESENVOLVIMENTO DE IMUNÓGENO CONJUGADO DE LIPOPOLISSACARÍDEO DE Pseudomonas Aeruginosa E TOXÓIDE TETÂNICO

2012 ◽  
Vol 17 (2) ◽  
Author(s):  
Bruno Gavinho ◽  
Rosangela Rodrigues Dos Santos ◽  
Luiz Felipe Caron ◽  
Maria Rita Sierakowski ◽  
Lucy Ono
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Clostridium Tetani

O lipopolissacarídeo (LPS) de Pseudomonas aeruginosa foi conjugado ao toxóide tetânico (TT, toxina inativada do Clostridium tetani) utilizando a técnica de aminação redutiva direta, para o desenvolvimento de uma vacina conjugada (LPS-TT). Avaliou-se a resposta imune in vivo após aplicações intraperitoneais do imunógeno LPS-TT em camundongos Swiss, administrando-se 4 aplicações de LPS-TT com intervalos entre as doses de 14 dias. Avaliou-se o título de anticorpos anti-Pseudomonas produzidos pela técnica de soroaglutinação microscópica. Os grupos que foram imunizados com LPS-TT e com LPS+TT (não conjugado quimicamente) apresentaram resposta de anticorpos aglutinantes superior ao controle imunizado apenas com LPS, e a resposta após o reforço vacinal dos animais imunizados com o conjugado LPS-TT foi 567% superior ao dos animais imunizados com LPS.

scholarly journals Tackling Virulence of Pseudomonas aeruginosa by the Natural Furanone Sotolon

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 871
Author(s):  
Mohammed F. Aldawsari ◽  
El-Sayed Khafagy ◽  
Ahmed Al Saqr ◽  
Ahmed Alalaiwe ◽  
Hisham A. Abbas ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Bacterial Infections ◽  
Therapeutic Agent ◽  
Bacterial Resistance ◽  
Inhibitory Concentration ◽  
Bacterial Biofilm ◽  
Resistant Bacteria ◽  
Resistance Development

The bacterial resistance development due to the incessant administration of antibiotics has led to difficulty in their treatment. Natural adjuvant compounds can be co-administered to hinder the pathogenesis of resistant bacteria. Sotolon is the prevailing aromatic compound that gives fenugreek its typical smell. In the current work, the anti-virulence activities of sotolon on Pseudomonas aeruginosa have been evaluated. P. aeruginosa has been treated with sotolon at sub-minimum inhibitory concentration (MIC), and production of biofilm and other virulence factors were assessed. Moreover, the anti-quorum sensing (QS) activity of sotolon was in-silico evaluated by evaluating the affinity of sotolon to bind to QS receptors, and the expression of QS genes was measured in the presence of sotolon sub-MIC. Furthermore, the sotolon in-vivo capability to protect mice against P. aeruginosa was assessed. Significantly, sotolon decreased the production of bacterial biofilm and virulence factors, the expression of QS genes, and protected mice from P. aeruginosa. Conclusively, the plant natural substance sotolon attenuated the pathogenicity of P. aeruginosa, locating it as a plausible potential therapeutic agent for the treatment of its infections. Sotolon can be used in the treatment of bacterial infections as an alternative or adjuvant to antibiotics to combat their high resistance to antibiotics.

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