Investigation on immunoprotectivity of recombinant vaccine candidates glutamine-binding periplasmic protein and putative peptidoglycan-binding protein in mouse model

Çiçek, Mustafa
Bordetella pertussis is a gram negative coccobacillus that causes pertussis known as whooping cough. After mass-vaccination started in 1940s, incidence of the disease has decreased. However, B. pertussis circulation in population has not been prevented completely. Starting from the first vaccination, development of several vaccines have been performed. These whole cell (Pw) and acellular pertussis (Pa) vaccines are not completely effective in terms of sustained, lifelong immunity and thus failure in eliminating subclinical infections poses a threat for both unimmunized infants and adults. The requirements for more effective acellular pertussis vaccines with protective proteins have raised the number of the studies to develop vaccines with high immune protective capacities. Recently, a surface antigen, namely glutamine-binding periplasmic protein GnlH was found to be among novel immunogenic proteins as shown by our immunoproteome group. Besides; another surface antigen putative peptidoglycan binding protein BP0020 was shown to include Lysm domain which is found in some of protective immunogens. These proteins were chosen as novel recombinant vaccine candidates. In the present study these proteins were tested for their immunoprotective capacity in mouse model. In order to stimulate humoral and cellular responses against B. pertussiss infection, the genes coding for gnlH and BP0020 proteins were amplified from the genomic DNA of B. pertussis strains and cloned into pGEM®-T Easy vector and sequenced. For their expression in Escherichia coli BL21(DE3) cells, expression vector pET-28a (+) was used together with IPTG induction system. After the expression of the desired proteins His-tag affinity chromatography together with dialysis was used for the purification of the proteins. Following Western blot analysis, 10 µg of both GlnlH and BP0020 proteins were used to immunize BALB/c mice (16-18 g) at days 0 and 21. B. pertussis Saadet live cells were then administered intranasally to challenge mice. Bacterial colonization in mice was determined after the removal of the lungs at days 5 and 8. When compared to the control groups, bacterial colonization were found to be decreased in the lungs of the mice immunized with 10 µg recombinant GlnH and BP0020 proteins. ELISA for serum-specific IgG levels was performed after the collection of sera. The results showed that IgG levels were significantly higher in immunized mice. In addition; serum IFN-γ levels were found to be higher in vaccinated mice in comparision to control groups.


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Tefon, Burcu Emine; Özcengiz, Gülay; Department of Biology (2012)
Bordetella pertussis is a gram-negative, human pathogen and etiologic agent of whooping cough (pertussis), a highly contagious, acute respiratory illness. In this study, the analysis of whole immunproteome and subproteomes of this microorganism was performed. The soluble cytoplasmic proteomes of B. pertussis Tohama I strain and a local isolate Saadet were separated by 2DE. By Western blot analysis, we identified 25 immunogenic proteins of three categories. In the first group, there were well-known proteins ...
A comprehensive analysis of Bordetella pertussis surface proteome and identification of new immunogenic proteins
Tefon, Burcu E.; Maass, Sandra; Ozcengiz, Erkan; Becher, Doerte; Hecker, Michael; Özcengiz, Gülay (2011-04-27)
Whooping cough, caused by the gram negative pathogen Bordetella pertussis, is a worldwide acute respiratory disease that predominantly involves infants. In the present study, surface proteins of B. pertussis Tohama I and Saadet strains were identified by using 2DE followed by MALDI-TOF-MS/MS analysis and also geLC-MS/MS. With these approaches it was possible to identify 45 and 226 proteins, respectively. When surface proteins of the strains were separated by 2DE and analyzed by Western blotting for their re...
Immune responses against the recombinant FimX and putative peptidyl-prolyl cis-trans isomerase from Bordetella pertussis
Yılmaz, Çiğdem; Özcengiz, Gülay; Department of Biology (2011)
Whooping cough (pertussis) is a highly contagious respiratory infection caused by Bordetella pertussis. It becomes widespread among adolescent and adults as well as infants. Although availability of effective pertussis vaccines seems to decrease the incidence of the disease, B. pertussis circulation in population has not been eliminated. It is thought that the antigenic drifts in major protective antigens and continued circulation of B. pertussis strains will result in gradual loss of the efficacy of the cu...
Immunoproteomic analysis of Bordetella pertussis and identification of new immunogenic proteins
Altindis, Emrah; Tefon, Burcu E.; Yildirim, Volkan; Ozcengiz, Erkan; Becher, Doete; Hecker, Michael; Özcengiz, Gülay (Elsevier BV, 2009-01-22)
Bordetella pertussis is the causative agent of highly communicable respiratory infection whooping cough (pertussis) which remains one of the world's leading causes of vaccine-preventable deaths. In the present study, total soluble proteins extracted from two B. pertussis strains, Tohama I and the local isolate Saadet were separated by two-dimensional gel electrophoresis and analyzed by Western blotting for their reactivity with the antisera obtained from the mice immunized with inactivated whole cells as we...
Assessment of immune protective capacities of the recombinant outer membrane protein q, iron superoxide dismutase and putative lipoprotein from bordetella pertussis
Yılmaz, Çiğdem; Özcengiz, Gülay; Department of Biology (2017)
Pertussis is a contagious disease which is commonly seen among infants and children and caused by a human pathogen known as Bordetella pertussis. There are currently two vaccine types available against the disease; whole-cell (wP) and acellular pertussis (aP) vaccines. Due to the side effects of wP vaccine, aP vaccines are commonly preferred for vaccination. Despite high vaccination coverage, high incidence rates among adolescents and adults have been reported causing the resurgence of pertussis to be the f...
Citation Formats
M. Çiçek, “Investigation on immunoprotectivity of recombinant vaccine candidates glutamine-binding periplasmic protein and putative peptidoglycan-binding protein in mouse model,” M.S. - Master of Science, Middle East Technical University, 2014.