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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

Sepharose CL-2B gel filtration profile of D-ALG conjugated to DT. Fractions were assayed for alginate at 210 nm and at 280 nm for DT.
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Figure 2: Sepharose CL-2B gel filtration profile of D-ALG conjugated to DT. Fractions were assayed for alginate at 210 nm and at 280 nm for DT.

Mentions: D-ALG-DT and D-LPS-DT were isolated by gel filtration, monitoring the elution profile by the UV absorbance at both 210 nm (for the presence of ALG and LPS) and 280 nm (for the presence of DT) and refractive index. Fractions with the maximum absorbance for both 210 and 280 nm (Fig. 2, fractions 72 to 77 and Fig. 3, fractions 72 to 74) were indicated the formation of D-ALG-DT and D-LPS-DT conjugate molecules. Various characteristics of conjugate vaccines are shown in Table 2. The conjugates were non-pyrogenic when tested at a dose of 50 μg/kg and evoked < 0.5°C increase in temperature after 24 h. The conjugates were non-toxic upon intraperitoneal administration to mice. No overt signs of illness and decrease in weight were observed and all mice survived. Mediums were observed after incubation at 37°C for 24 to 48 h. No signs of microorganism growth were observed. Sterility testing showed that the resulting conjugates were sterile. The above results, demonstrating the safety and stability of the conjugate vaccines, led us to evaluate its acceptability and immunogenicity in animals.


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)

Sepharose CL-2B gel filtration profile of D-ALG conjugated to DT. Fractions were assayed for alginate at 210 nm and at 280 nm for DT.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: Sepharose CL-2B gel filtration profile of D-ALG conjugated to DT. Fractions were assayed for alginate at 210 nm and at 280 nm for DT.
Mentions: D-ALG-DT and D-LPS-DT were isolated by gel filtration, monitoring the elution profile by the UV absorbance at both 210 nm (for the presence of ALG and LPS) and 280 nm (for the presence of DT) and refractive index. Fractions with the maximum absorbance for both 210 and 280 nm (Fig. 2, fractions 72 to 77 and Fig. 3, fractions 72 to 74) were indicated the formation of D-ALG-DT and D-LPS-DT conjugate molecules. Various characteristics of conjugate vaccines are shown in Table 2. The conjugates were non-pyrogenic when tested at a dose of 50 μg/kg and evoked < 0.5°C increase in temperature after 24 h. The conjugates were non-toxic upon intraperitoneal administration to mice. No overt signs of illness and decrease in weight were observed and all mice survived. Mediums were observed after incubation at 37°C for 24 to 48 h. No signs of microorganism growth were observed. Sterility testing showed that the resulting conjugates were sterile. The above results, demonstrating the safety and stability of the conjugate vaccines, led us to evaluate its acceptability and immunogenicity in animals.

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