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Immunogenicity comparison of conjugate vaccines composed of alginate and lipopolysaccharide of Pseudomonas aeruginosa bound to diphtheria toxoid.

Najafzadeh F, Jaberi G, Shapouri R, Rahnema M, Karimi-Nik A, Kianmehr A - Iran J Microbiol (2014)

Bottom Line: The resulting depolymerized alginate (D-ALG) and detoxified LPS (D-LPS) were covalently coupled to diphtheria toxoid (DT) as a carrier protein with adipic acid dihydrazide (ADH) as a spacer molecule and carbodiimide as a linker.The conjugates were non-toxic and non-pyrogenic.ELISA results indicated that antibodies titer of D-ALGDT was more than D-LPSDT.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran.

ABSTRACT

Background and objectives: Treatment of Pseudomonas aeruginosa infections is greatly hampered by innate and acquired antibiotic resistance. The goal of this study was to compure the immunogenicity of conjugates of P. aeruginosa depolymerized alginate-diphtheria toxoid (D-ALGDT) and P. aeruginosa detoxified lipopolysaccharidediphtheria toxoid (D-LPSDT) in mouse model.

Materials and methods: Alginate and LPS were purified from P. aeruginosa strain PAO1. The resulting depolymerized alginate (D-ALG) and detoxified LPS (D-LPS) were covalently coupled to diphtheria toxoid (DT) as a carrier protein with adipic acid dihydrazide (ADH) as a spacer molecule and carbodiimide as a linker. Sterility, safety and pyrogenicity tests were performed. 30 mice in two groups were immunized intraperitoneally on days 0, 14 and 28 with 10 μg of D-ALGDT and D-LPSDT. Conjugates specific antibody levels were also determined by enzyme-linked immunosorbent assay (ELISA).

Results: The conjugates were non-toxic and non-pyrogenic. Conjugates of D-ALGDT and D-LPSDT were shown to be safe and to elicit total IgG, IgM, IgA, IgG1, IgG2a, IgG2b and IgG3 antibodies in mice. ELISA results indicated that antibodies titer of D-ALGDT was more than D-LPSDT.

Conclusion: Immunization with D-ALGDT showed significant increase in all types of antibodies titers in versus D-LPSDT, suggesting D-ALGDT as a vaccine candidate against P. aeruginosa infections.

No MeSH data available.


Related in: MedlinePlus

Silver-stained SDS-PAGE in 14% gel of P. aeruginosa LPS. Lane 1 and 2 were loaded with 10 μg/ml and 5 μg/ml LPS, respectively.
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Figure 1: Silver-stained SDS-PAGE in 14% gel of P. aeruginosa LPS. Lane 1 and 2 were loaded with 10 μg/ml and 5 μg/ml LPS, respectively.

Mentions: Fig. 1 shows the electrophoretic patterns of the LPS bands of P. aeruginosa on an SDS-PAGE. Various characteristics of D-ALG and D-LPS were also shown in Table 1. As can be found from the Table 1, the endotoxin activity assay showed 0.125 EU/ml of D-ALG and D-LPS that were acceptable to use for immunization. D-ALG and D-LPS were non-pyrogenic when tested at a dose of 50 μg/kg and evoked < 0.5°C increase in temperature after 24 h.


Immunogenicity comparison of conjugate vaccines composed of alginate and lipopolysaccharide of Pseudomonas aeruginosa bound to diphtheria toxoid.

Najafzadeh F, Jaberi G, Shapouri R, Rahnema M, Karimi-Nik A, Kianmehr A - Iran J Microbiol (2014)

Silver-stained SDS-PAGE in 14% gel of P. aeruginosa LPS. Lane 1 and 2 were loaded with 10 μg/ml and 5 μg/ml LPS, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4385571&req=5

Figure 1: Silver-stained SDS-PAGE in 14% gel of P. aeruginosa LPS. Lane 1 and 2 were loaded with 10 μg/ml and 5 μg/ml LPS, respectively.
Mentions: Fig. 1 shows the electrophoretic patterns of the LPS bands of P. aeruginosa on an SDS-PAGE. Various characteristics of D-ALG and D-LPS were also shown in Table 1. As can be found from the Table 1, the endotoxin activity assay showed 0.125 EU/ml of D-ALG and D-LPS that were acceptable to use for immunization. D-ALG and D-LPS were non-pyrogenic when tested at a dose of 50 μg/kg and evoked < 0.5°C increase in temperature after 24 h.

Bottom Line: The resulting depolymerized alginate (D-ALG) and detoxified LPS (D-LPS) were covalently coupled to diphtheria toxoid (DT) as a carrier protein with adipic acid dihydrazide (ADH) as a spacer molecule and carbodiimide as a linker.The conjugates were non-toxic and non-pyrogenic.ELISA results indicated that antibodies titer of D-ALGDT was more than D-LPSDT.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran.

ABSTRACT

Background and objectives: Treatment of Pseudomonas aeruginosa infections is greatly hampered by innate and acquired antibiotic resistance. The goal of this study was to compure the immunogenicity of conjugates of P. aeruginosa depolymerized alginate-diphtheria toxoid (D-ALGDT) and P. aeruginosa detoxified lipopolysaccharidediphtheria toxoid (D-LPSDT) in mouse model.

Materials and methods: Alginate and LPS were purified from P. aeruginosa strain PAO1. The resulting depolymerized alginate (D-ALG) and detoxified LPS (D-LPS) were covalently coupled to diphtheria toxoid (DT) as a carrier protein with adipic acid dihydrazide (ADH) as a spacer molecule and carbodiimide as a linker. Sterility, safety and pyrogenicity tests were performed. 30 mice in two groups were immunized intraperitoneally on days 0, 14 and 28 with 10 μg of D-ALGDT and D-LPSDT. Conjugates specific antibody levels were also determined by enzyme-linked immunosorbent assay (ELISA).

Results: The conjugates were non-toxic and non-pyrogenic. Conjugates of D-ALGDT and D-LPSDT were shown to be safe and to elicit total IgG, IgM, IgA, IgG1, IgG2a, IgG2b and IgG3 antibodies in mice. ELISA results indicated that antibodies titer of D-ALGDT was more than D-LPSDT.

Conclusion: Immunization with D-ALGDT showed significant increase in all types of antibodies titers in versus D-LPSDT, suggesting D-ALGDT as a vaccine candidate against P. aeruginosa infections.

No MeSH data available.


Related in: MedlinePlus