title: Streptococcus pneumoniae fructose-1,6-bisphosphate aldolase, a protein vaccine candidate, elicits Th1/Th2/Th17-type cytokine responses in mice.
abstract: Streptococcus pneumoniae (S. pneumoniae) is a major pathogen worldwide. The currently available polysaccharide-based vaccines significantly reduce morbidity and mortality. However, the inherent disadvantages of the currently available polysaccharide-based vaccines have motivated the search for other bacterial immunogens capable of eliciting a protective immune response against S. pneumoniae. Fructose-1,6-bisphosphate aldolase (FBA) is a glycolytic enzyme, which was found to localize to the bacterial surface, where it functions as an adhesin. Previously, immunizing mice with recombinant FBA (rFBA) in the presence of alum elicited a protective immune response against a lethal challenge with S. pneumoniae. Thus, the aim of the present study was to determine the cytokine responses that are indicative of protective immunity following immunization with rFBA. The protective effects against pneumococcal challenge in mice immunized with rFBA with complete Freund's adjuvant (CFA) in the initial immunization and with incomplete Freund's adjuvant (IFA) in booster immunizations surpassed the protective effects observed following immunization with either rFBA + alum or pVACfba. CD4+ T-cells obtained from the rFBA/CFA/IFA/IFA-immunized mice co-cultured with rFBA-pulsed antigen-presenting cells (APCs), exhibited a significantly greater proliferative ability than CD4+ T-cells obtained from the adjuvant-immunized mice co-cultured with rFBA‑pulsed APCs. The levels of the Th1-type cytokines, interferon (IFN)-γ, interleukin (IL)-2, tumor necrosis factor (TNF)-α and IL-12, the Th2-type cytokines, IL-4, IL-5 and IL-10, and the Th17-type cytokine, IL-17A, significantly increased within 72 h of the initiation of co-culture with CD4+ T-cells obtained from the rFBA‑immunized mice, in comparison with the co-cultures with CD4+ T-cells obtained from the adjuvant-immunized mice. Immunizing mice with rFBA resulted in an IgG1/IgG2 ratio of 41, indicating a Th2 response with substantial Th1 involvement. In addition, rabbit and mouse anti-rFBA antisera significantly protected the mice against a lethal S. pneumoniae challenge in comparison with preimmune sera. Our results emphasize the mixed involvement of the Th1, Th2 and Th17 arms of the immune system in response to immunization with pneumococcal rFBA, a potential vaccine candidate.
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Shirin
authors.initials: S
authors.lastname: Elhaik Goldman
authors.affiliation: NasVax/Protea Vaccine Technologies Ltd., Kiryat Weizmann, Science Park, Ness Ziona 74140, Israel.
authors.forename: Shahar
authors.initials: S
authors.lastname: Dotan
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Amir
authors.initials: A
authors.lastname: Talias
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Lilo
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Shalhevet
authors.initials: S
authors.lastname: Azriel
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Itay
authors.initials: I
authors.lastname: Malka
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Maxim
authors.initials: M
authors.lastname: Portnoi
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Ariel
authors.initials: A
authors.lastname: Ohayon
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kafka
authors.affiliation: NasVax/Protea Vaccine Technologies Ltd., Kiryat Weizmann, Science Park, Ness Ziona 74140, Israel.
authors.forename: Ronald
authors.initials: R
authors.lastname: Ellis
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Moshe
authors.initials: M
authors.lastname: Elkabets
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Angel
authors.initials: A
authors.lastname: Porgador
authors.affiliation: Prof. Ephraim Katzir Department of Biotechnology Engineering, ORT Braude College, Karmiel 21982, Israel.
authors.forename: Ditza
authors.initials: D
authors.lastname: Levin
authors.affiliation: Prof. Ephraim Katzir Department of Biotechnology Engineering, ORT Braude College, Karmiel 21982, Israel.
authors.forename: Rosa
authors.initials: R
authors.lastname: Azhari
authors.affiliation: Division of Infectious Diseases, University of Mississippi Medical Center, Jackson, MS 39216, USA.
authors.forename: Edwin
authors.initials: E
authors.lastname: Swiatlo
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Eduard
authors.initials: E
authors.lastname: Ling
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Galia
authors.initials: G
authors.lastname: Feldman
authors.affiliation: NasVax/Protea Vaccine Technologies Ltd., Kiryat Weizmann, Science Park, Ness Ziona 74140, Israel.
authors.forename: Michael
authors.initials: M
authors.lastname: Tal
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Ron
authors.initials: R
authors.lastname: Dagan
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center, Beer‑Sheva 84100, Israel.
authors.forename: Yaffa
authors.initials: Y
authors.lastname: Mizrachi Nebenzahl
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D016207:Cytokines / Q000276:immunology*; D005260:Female; D005620:Freund's Adjuvant / Q000276:immunology / Q000627:therapeutic use; D005634:Fructose-Bisphosphate Aldolase / Q000276:immunology / Q000627:therapeutic use*; D007114:Immunization; D008055:Lipids / Q000276:immunology / Q000627:therapeutic use; D051379:Mice; D008807:Mice, Inbred BALB C; D011008:Pneumococcal Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D011817:Rabbits; D022541:Streptococcal Vaccines / Q000276:immunology / Q000627:therapeutic use*; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology* / Q000382:microbiology; D018417:Th1 Cells / Q000276:immunology; D058504:Th17 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; D008055:Lipids; D022541:Streptococcal Vaccines; C114843:incomplete Freund's adjuvant; D005620:Freund's Adjuvant; D005634:Fructose-Bisphosphate Aldolase