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Hmw1555-914, Hmw2553-916, and Hia585-705As Subunit Vaccine Candidates of Nontypeable Haemophilus Influenzae Induce Specific Antibody Responses With Bactericidal Activity in Balb/C Publisher



Abbaszadehgoudarzi K1, 2 ; Abbaszadehgoudarzi G3, 4 ; Khorramizadeh MR5 ; Bouzari S6 ; Rezaie S7 ; Davari M2, 8 ; Siadat SD2, 8 ; Teimooritoolabi L1, 9
Authors
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Authors Affiliations
  1. 1. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  3. 3. Department of Medical Biotechnology, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
  4. 4. Cancer Prevention Research Center, Shahroud University of Medical Sciences, Shahroud, Iran
  5. 5. Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
  7. 7. Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  8. 8. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran
  9. 9. Molecular Medicine Department, Pasteur Institute of Iran, Tehran, Iran

Source: Jundishapur Journal of Microbiology Published:2018


Abstract

Background: Nontypeable Haemophilus influenzae (NTHi) is responsible for diseases such as otitis media, sinusitis, bronchitis, and pneumonia. Unlike H. influenzae Serotype b (Hib), there is no vaccine against diseases induced by NTHi. High molecular weight1 (HMW1), high molecular weight 2 (HMW2), and H. influenzae type a (Hia) proteins are critical protein adhesions of NTHi with the potentiality to provide the protection. Objectives: The current study aimed at investigating the potential of recombinant HMW1555-914, HMW2553-916, and Hia585-705proteins as subunit vaccines to induce immune responses after active immunization in Bagg albino (inbred research mouse strain) or Balb/c mice. Methods: HMW1555-914, HMW2553-916, and Hia585-705amplified by polymerase chain reaction (PCR) were cloned into pET28a. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and theWestern blotting analysis were carried out to investigate the characteristics of proteins. Proteins were purified by Ni-nitrilotriacetic acid (Ni-NTA) as a gel matrix. Mice (Balb/c) were immunized subcutaneously with HMW1555-914, HMW2553-916, and Hia585-705proteins, alone or in binary and ternary combinations. Eventually, specific antibody responses were evaluated by the enzyme-linked immunosorbent assay (ELISA) and serum bactericidal assay (SBA). Results: Antibody responses significantly increased in Balb/cimmunizedby ternary combinationcomparedwith the controlgroup. IgG1/IgG2a ratio indicated that proteins directed immune responses toward T-helper 2. Antisera produced against purified proteins in ternary combination showed the highest bactericidal activity against NTHi strains with the titer of 1:32. Conclusions: The obtained results suggested that HMW1555-914, HMW2553-916, and Hia585-705adhesins were the potential subunit vaccine candidates of NTHi and after further investigation could be considered for protection against NTHi infections. © 2017, Jundishapur Journal of Microbiology.