title: Preparation of Pseudomonas aeruginosa alginate-flagellin immunoconjugate.
abstract: Mucoid strains of Pseudomonas aeruginosa are closely associated with chronic pulmonary infections. In this report we describe a straightforward approach to conjugate high molecular weight alginate to type b-flagellin (FLB) and investigation of its bioactivity. The conjugation process was performed by using ADH and EDAC. The endotoxin was eliminated from the candidate vaccine by LPS removal resin followed by LAL test. The bioconjugate molecules were verified by simultaneously determination of polysaccharide/protein content followed by gel filtration chromatography and FTIR spectroscopy. Groups of eight BALB/c mice were injected intranasally with 5 μg (per each nostril) of purified alginate, FLB and conjugated alginate-FLB with two week intervals. The functional activity of the vaccine was evaluated by ELISA and opsonophagocytosis tests. Vaccination with the alginate-FLB conjugate induced a significant (P = 0.0033) rise in alginate specific IgG in mice. At all dilution ranges, the opsonic activity of the conjugate vaccine antisera was significantly higher than alginate alone (61.9% vs. 17.3% at 1:4 dilution; P = 0.0067). The alginate-FLB conjugate could elicit high specific antibodies titer against alginate by improving its immunogenicity. In addition, the antisera raised against conjugate vaccine act as a suitable opsonin for phagocytosis of the mucoid strains of P. aeruginosa.
authors.affiliation: Departments of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran; Mycobacteriology Research Center (MRS), Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Sobhan
authors.initials: S
authors.lastname: Faezi
authors.affiliation: Departments of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran; Mycobacteriology Research Center (MRS), Pasteur Institute of Iran, Tehran, Iran. Electronic address: Padideh79@yahoo.com.
authors.forename: Ahmad Reza
authors.initials: AR
authors.lastname: Bahrmand
authors.affiliation: Departments of Immunology, Pasteur Institute of Iran, Tehran, Iran. Electronic address: Mahdavivac@gmail.com.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mahdavi
authors.affiliation: Departments of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran; Mycobacteriology Research Center (MRS), Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Davar
authors.initials: SD
authors.lastname: Siadat
authors.affiliation: Biotechnology Research Center, Drug Design and Bioinformatics Group, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Soroush
authors.initials: S
authors.lastname: Sardari
authors.affiliation: Medical Biotechnology Research Center, Faculty of Paramedicine, Guilan University of Medical Sciences, Rasht, Iran.
authors.forename: Iraj
authors.initials: I
authors.lastname: Nikokar
authors.affiliation: Department of Clinical Biochemistry, Medical Biotechnology Research Center, Faculty of Paramedicine, Guilan University of Medical Sciences, Rasht, Iran.
authors.forename: Korosh
authors.initials: K
authors.lastname: Khanaki
authors.affiliation: Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
authors.forename: Ebrahim
authors.initials: E
authors.lastname: Mirzajani
authors.affiliation: School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Goudarzi
mesh_terms: D000818:Animals; D005260:Female; D005408:Flagellin / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D018796:Immunoconjugates / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D011552:Pseudomonas Infections / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D054406:Pseudomonas Vaccines / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000276:immunology
chemical_list: D018796:Immunoconjugates; D054406:Pseudomonas Vaccines; D005408:Flagellin
keywords: Alginate; Conjugation; Flagellin; Pseudomonas aeruginosa