title: Recombinant C-terminal 311 amino acids of HapS adhesin as a vaccine candidate for nontypeable Haemophilus influenzae: A study on immunoreactivity in Balb/C mouse.
abstract: Hap, an auto-transporter protein, is an antigenically conserved adhesion protein which is present on both typeable and nontypeable Haemophilus influenzae. This protein has central role in bacterial attachment to respiratory tract epithelial cells. A 1000bp C-terminal fragment of Hap passenger domain (HapS) from nontypeable Haemophilus influenzae was cloned into a prokaryotic expression vector, pET-24a. BALB/c mice were immunized subcutaneously with purified rC-HapS. Serum IgG responses to purified rC-HapS, serum IgG subclasses were determined by ELISA and functional activity of antibodies was examined by Serum Bactericidal Assay. The output of rC-HapS was approximately 62% of the total bacterial proteins. Serum IgG responses were significantly increased in immunized group with rC-HapS mixed with Freund's adjuvant in comparison with control groups. Analysis of the serum IgG subclasses showed that the IgG1 subclass was predominant after subcutaneous immunization in BALB/c mice (IgG2a/IgG1 < 1). The sera from rC-HapS immunized animals were strongly bactericidal against nontypeable Haemophilus influenzae. These results suggest that rC-HapS may be a potential vaccine candidate for nontypeable Haemophilus influenzae.
authors.affiliation: Department of Biology, Faculty of Basic Sciences, Islamic Azad University of Ghiamdasht (East Tehran Branch), Tehran, Iran. Electronic address: akram_tabatabaee@yahoo.com.
authors.forename: Akram Sadat
authors.initials: AS
authors.lastname: Tabatabaee Bafroee
authors.affiliation: Department of Mycobacteriology & Pulmonary Research, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran. Electronic address: d.siadat@gmail.com.
authors.forename: Seyed Davar
authors.initials: SD
authors.lastname: Siadat
authors.affiliation: Department of Bacteriology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Fazlollah
authors.initials: SF
authors.lastname: Mousavi
authors.affiliation: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Aghasadeghi
authors.affiliation: Iranian Blood Research and Fractionation Holding Company, Tehran, Iran.
authors.forename: Hashem
authors.initials: H
authors.lastname: Khorsand
authors.affiliation: Department of Bacteriology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Nejati
authors.affiliation: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Mehdi
authors.initials: SM
authors.lastname: Sadat
authors.affiliation: Department of Virology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mahdavi
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001770:Blood Bactericidal Activity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005620:Freund's Adjuvant / Q000008:administration & dosage; D015870:Gene Expression; D005822:Genetic Vectors; D018073:Haemophilus Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D006193:Haemophilus influenzae / Q000235:genetics / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D007279:Injections, Subcutaneous; D008807:Mice, Inbred BALB C; D050296:Microbial Viability; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D018073:Haemophilus Vaccines; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D005620:Freund's Adjuvant
keywords: IgG subclasses; Nontypeable Haemophilus influenzae; Serum bactericidal assay; rC-HapS