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  • 1.
    Khalaf, Hazem
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Nakka, Sravya Sowdamini
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden. PEAS Institut AB, Söderleden 1, Linköping.
    Sandén, Camilla
    Division of Molecular Physics, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
    Svärd, Anna
    Division of Molecular Physics, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
    Hultenby, Kjell
    Division of Clinical Research Centre, Department of Laboratory Medicine, Karolinska.
    Scherbak, Nikolai
    Örebro University, School of Science and Technology.
    Aili, Daniel
    PEAS Institut AB, Söderleden 1, Linköping.
    Bengtsson, Torbjörn
    Örebro University, School of Medicine, Örebro University, Sweden.
    Antibacterial effects of lactobacillus and bacteriocin NC8 αβ on the periodontal pathogen Porphyromonas gingivalisManuscript (preprint) (Other academic)
  • 2.
    Khalaf, Hazem
    et al.
    Örebro University, School of Medical Sciences.
    Nakka, Sravya Sowdamini
    Örebro University, School of Medical Sciences. PEAS Institut AB, Linköping, Sweden.
    Sandén, Camilla
    Division of Molecular Physics, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
    Svärd, Anna
    Division of Molecular Physics, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
    Hultenby, Kjell
    Division of Clinical Research Centre, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.
    Scherbak, Nikolai
    Örebro University, School of Science and Technology.
    Aili, Daniel
    Division of Molecular Physics, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
    Bengtsson, Torbjörn
    Örebro University, School of Medical Sciences.
    Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis2016In: BMC Microbiology, ISSN 1471-2180, E-ISSN 1471-2180, Vol. 16, no 1, article id 188Article in journal (Refereed)
    Abstract [en]

    Background: The complications in healthcare systems associated with antibiotic-resistant microorganisms have resulted in an intense search for new effective antimicrobials. Attractive substances from which novel antibiotics may be developed are the bacteriocins. These naturally occurring peptides are generally considered to be safe and efficient at eliminating pathogenic bacteria. Among specific keystone pathogens in periodontitis, Porphyromonas gingivalis is considered to be the most important pathogen in the development and progression of chronic inflammatory disease. The aim of the present study was to investigate the antimicrobial effects of different Lactobacillus species and the two-peptide bacteriocin PLNC8 αβ on P. gingivalis.

    Results: Growth inhibition of P. gingivalis was obtained by viable Lactobacillus and culture media from L. plantarum NC8 and 44048, but not L. brevis 30670. The two-peptide bacteriocin from L. plantarum NC8 (PLNC8 αβ) was found to be efficient against P. gingivalis through binding followed by permeabilization of the membranes, using Surface plasmon resonance analysis and DNA staining with Sytox Green. Liposomal systems were acquired to verify membrane permeabilization by PLNC8 αβ. The antimicrobial activity of PLNC8 αβ was found to be rapid (1 min) and visualized by TEM to cause cellular distortion through detachment of the outer membrane and bacterial lysis.

    Conclusion: Soluble or immobilized PLNC8 αβ bacteriocins may be used to prevent P. gingivalis colonization and subsequent pathogenicity, and thus supplement the host immune system against invading pathogens associated with periodontitis.

  • 3.
    Nakka, Sravya Sowdamini
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Development of novel tools for prevention and diagnosis of Porphyromonas gingivalis infection and periodontitis 2015Licentiate thesis, comprehensive summary (Other academic)
    Abstract [en]

    Periodontitis is a chronic infectious disease causing inflammation and destruction of tooth-supporting structures eventually leading to tooth loss. Periodontitis is induced by different subgingival microorganisms in combination with an excessive and dysregulated immune-inflammatory response against this microbial challenge. The pathogenicity of the periodontal biofilm is highly dependent upon some "keystone" species of which Porphyromonas gingivalis is considered to be one of the most important pathogens. P. gingivalis expresses a broad range of virulence factors including lipopolysaccharides (LPS), fimbriae, adhesins, hemagglutinins and an array of proteolytic enzymes. Among these factors, cysteine proteases (gingipains) are of special importance both for the bacterial survival/proliferation and for the pathological outcome. Gingipains consist of arginine-specific proteases (RgpA and RgpB) and lysine-specific protease (Kgp). The major aim of this thesis was to develop and test novel methods for diagnosis and prevention of P. gingivalis infection and periodontitis. In study I, host lymphocytes were stimulated with P. gingivalis to develop antibodies in vitro and the anti-P. gingivalis antibodies were used for immunodetection in clinical samples. The in vitro method of antibody production developed during this study could be used for an efficient real-time detection of P. gingivalis infection and periodontitis, while the attenuating effects of the antibodies suggest a role in passive immunization to prevent periodontitis and its associated diseases. In study II, we have elucidated the properties and antimicrobial effects of different lactobacillus species and the two-peptide bacteriocin NC8 αβ on P. gingivalis. NC8 αβ was found to be efficient against P. gingivalis through bacterial binding followed by permeabilization of the membranes. Liposomal systems were acquired to verify membrane permeabilization by NC8 αβ. The antimicrobial activity of NC8 αβ was found to be rapid, potent and instant. In conclusion, soluble or immobilized NC8 αβ bacteriocins may be used to prevent P. gingivalis colonization and subsequent pathogenicity, and thus supplement the host immune system against invading pathogens associated with periodontitis.

    List of papers
    1. Antibodies produced in vitro in the detection of periodontal bacteria by using surface plasmon resonance analysis
    Open this publication in new window or tab >>Antibodies produced in vitro in the detection of periodontal bacteria by using surface plasmon resonance analysis
    Show others...
    2015 (English)In: Clinical and Experimental Dental Research, ISSN 2057-4347, Vol. 1, no 1, p. 32-44Article in journal (Refereed) Published
    Abstract [en]

    Porphyromonas gingivalis (P. gingivalis) is a major etiological agent associated with periodontitis. This study aims to develop antibodies to P. gingivalis in vitro for real-time detection of bacteria in clinical samples. Lymphocytes were isolated from whole blood of patient treated for periodontitis and were stimulated with P. gingivalis ATCC 33277. B-cell maturation to long-living antibody secreting-plasma cells was studied using flow cytometry and immunofluorescence staining. The antibodies developed in vitro were immobilized onto a CM-5 sensor chip of a biosensor to detect the presence of P. gingivalis in the gingival crevicular fluid of patients with periodontitis compared to periodontally healthy controls (n = 30). Surface plasmon resonance (SPR) analysis was performed to evaluate specific interactions of bacteria in samples with the immobilized antibodies. The results of SPR analysis were compared to the detection of P. gingivalis in the samples using DNA–DNA checkerboard hybridization technique. A clear and distinct change in lymphocyte morphology upon stimulation with P. gingivalis was observed. Anti-P. gingivalis antibodies secreted by CD38+ plasma cells showed the presence of all the four IgG subclasses. The results of DNA–DNA checkerboard analysis were in agreement with that of SPR analysis for the detection of P. gingivalis in patient samples. Furthermore, incubation with anti-P. gingivalis attenuated the bacterial response in SPR. The in vitro method for antibody production developed during this study could be used for an efficient real-time detection of periodontitis, and the attenuating effects of in vitro antibodies suggest their role in passive immunization to prevent periodontitis and their associated risk factors.

    Place, publisher, year, edition, pages
    Wiley-Blackwell, 2015
    Keywords
    Antibodies, B-cell, plasma cell, periodontitis, P. gingivalis, surface plasmon resonance
    National Category
    Microbiology in the medical area
    Research subject
    Biomedicine
    Identifiers
    urn:nbn:se:oru:diva-46728 (URN)10.1002/cre2.6 (DOI)000216889200006 ()2-s2.0-84986898105 (Scopus ID)
    Funder
    Knowledge Foundation
    Available from: 2015-11-24 Created: 2015-11-23 Last updated: 2018-04-07Bibliographically approved
    2. Antibacterial effects of lactobacillus and bacteriocin NC8 αβ on the periodontal pathogen Porphyromonas gingivalis
    Open this publication in new window or tab >>Antibacterial effects of lactobacillus and bacteriocin NC8 αβ on the periodontal pathogen Porphyromonas gingivalis
    Show others...
    (English)Manuscript (preprint) (Other academic)
    National Category
    Microbiology in the medical area
    Research subject
    Biomedicine
    Identifiers
    urn:nbn:se:oru:diva-46730 (URN)
    Available from: 2015-11-24 Created: 2015-11-24 Last updated: 2018-01-10Bibliographically approved
  • 4.
    Nakka, Sravya Sowdamini
    Örebro University, School of Medical Sciences.
    Development of novel tools for prevention and diagnosis of Porphyromonas gingivalis infection and periodontitis2016Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Periodontitis is a chronic inflammatory disease caused by exaggerated host immune responses to dysregulated microbiota in dental biofilms leading to degradation of tissues and alveolar bone loss. Porphyromonas gingivalis is a major periodontal pathogen and expresses several potent virulence factors. Among these factors, arginine and lysine gingipains are of special importance, both for the bacterial survival/proliferation and the pathological outcome. The major aim of this thesis was to develop and test novel methods for diagnosis and prevention of P. gingivalis infection and periodontitis. In study I, anti-P. gingivalis antibodies were developed in vitro for immunodetection of bacteria in clinical samples using a surface plasmon resonance (SPR)-based biosensor. Specific binding of the antibodies to P. gingivalis was demonstrated in samples of patients with periodontitis and the results were validated using real-time PCR and DNA-DNA checkerboard analysis. In study II, we elucidated the properties and antimicrobial effects of different lactobacillus species and the two-peptide bacteriocin PLNC8 αβ on P. gingivalis. L. plantarum NC8 and 44048 effectively inhibited P. gingivalis growth and pure PLNC8 αβ induced bacterial lysis by damaging P. gingivalis membrane. In study III, we demonstrated that PLNC8 αβ dose-dependently induces proliferation and release of growth factors in gingival epithelial cells (GECs). Furthermore, PLNC8 αβ decreased P. gingivalis-induced cytotoxic effects in GECs but did not alter the effect of gingipains on cytokine expression. In study IV, we elucidated the effects of anti-P. gingivalis antibodies and PLNC8 αβ in regulating cellular responses during P. gingivalis infection. Both antibodies and PLNC8 αβ modulated P. gingivalis-induced expression of growth factors in GECs, however, their effects were diminished when used in combination. The results of this thesis demonstrate a possible role of anti-P. gingivalis antibodies and PLNC8 αβ in prevention and treatment of P. gingivalis infection and periodontitis with no cytotoxic effects on human cells.

    List of papers
    1. Antibodies produced in vitro in the detection of periodontal bacteria by using surface plasmon resonance analysis
    Open this publication in new window or tab >>Antibodies produced in vitro in the detection of periodontal bacteria by using surface plasmon resonance analysis
    Show others...
    2015 (English)In: Clinical and Experimental Dental Research, ISSN 2057-4347, Vol. 1, no 1, p. 32-44Article in journal (Refereed) Published
    Abstract [en]

    Porphyromonas gingivalis (P. gingivalis) is a major etiological agent associated with periodontitis. This study aims to develop antibodies to P. gingivalis in vitro for real-time detection of bacteria in clinical samples. Lymphocytes were isolated from whole blood of patient treated for periodontitis and were stimulated with P. gingivalis ATCC 33277. B-cell maturation to long-living antibody secreting-plasma cells was studied using flow cytometry and immunofluorescence staining. The antibodies developed in vitro were immobilized onto a CM-5 sensor chip of a biosensor to detect the presence of P. gingivalis in the gingival crevicular fluid of patients with periodontitis compared to periodontally healthy controls (n = 30). Surface plasmon resonance (SPR) analysis was performed to evaluate specific interactions of bacteria in samples with the immobilized antibodies. The results of SPR analysis were compared to the detection of P. gingivalis in the samples using DNA–DNA checkerboard hybridization technique. A clear and distinct change in lymphocyte morphology upon stimulation with P. gingivalis was observed. Anti-P. gingivalis antibodies secreted by CD38+ plasma cells showed the presence of all the four IgG subclasses. The results of DNA–DNA checkerboard analysis were in agreement with that of SPR analysis for the detection of P. gingivalis in patient samples. Furthermore, incubation with anti-P. gingivalis attenuated the bacterial response in SPR. The in vitro method for antibody production developed during this study could be used for an efficient real-time detection of periodontitis, and the attenuating effects of in vitro antibodies suggest their role in passive immunization to prevent periodontitis and their associated risk factors.

    Place, publisher, year, edition, pages
    Wiley-Blackwell, 2015
    Keywords
    Antibodies, B-cell, plasma cell, periodontitis, P. gingivalis, surface plasmon resonance
    National Category
    Microbiology in the medical area
    Research subject
    Biomedicine
    Identifiers
    urn:nbn:se:oru:diva-46728 (URN)10.1002/cre2.6 (DOI)000216889200006 ()2-s2.0-84986898105 (Scopus ID)
    Funder
    Knowledge Foundation
    Available from: 2015-11-24 Created: 2015-11-23 Last updated: 2018-04-07Bibliographically approved
    2. Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis
    Open this publication in new window or tab >>Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis
    Show others...
    2016 (English)In: BMC Microbiology, ISSN 1471-2180, E-ISSN 1471-2180, Vol. 16, no 1, article id 188Article in journal (Refereed) Published
    Abstract [en]

    Background: The complications in healthcare systems associated with antibiotic-resistant microorganisms have resulted in an intense search for new effective antimicrobials. Attractive substances from which novel antibiotics may be developed are the bacteriocins. These naturally occurring peptides are generally considered to be safe and efficient at eliminating pathogenic bacteria. Among specific keystone pathogens in periodontitis, Porphyromonas gingivalis is considered to be the most important pathogen in the development and progression of chronic inflammatory disease. The aim of the present study was to investigate the antimicrobial effects of different Lactobacillus species and the two-peptide bacteriocin PLNC8 αβ on P. gingivalis.

    Results: Growth inhibition of P. gingivalis was obtained by viable Lactobacillus and culture media from L. plantarum NC8 and 44048, but not L. brevis 30670. The two-peptide bacteriocin from L. plantarum NC8 (PLNC8 αβ) was found to be efficient against P. gingivalis through binding followed by permeabilization of the membranes, using Surface plasmon resonance analysis and DNA staining with Sytox Green. Liposomal systems were acquired to verify membrane permeabilization by PLNC8 αβ. The antimicrobial activity of PLNC8 αβ was found to be rapid (1 min) and visualized by TEM to cause cellular distortion through detachment of the outer membrane and bacterial lysis.

    Conclusion: Soluble or immobilized PLNC8 αβ bacteriocins may be used to prevent P. gingivalis colonization and subsequent pathogenicity, and thus supplement the host immune system against invading pathogens associated with periodontitis.

    Place, publisher, year, edition, pages
    London, United Kingdom: BioMed Central, 2016
    Keywords
    Periodontitis, P. gingivalis, Lactobacillus, Bacteriocin, PLNC8
    National Category
    Microbiology
    Identifiers
    urn:nbn:se:oru:diva-51749 (URN)10.1186/s12866-016-0810-8 (DOI)000383422500001 ()27538539 (PubMedID)2-s2.0-84982308370 (Scopus ID)
    Funder
    Swedish Heart Lung FoundationKnowledge FoundationMagnus Bergvall Foundation
    Note

    Funding Agency:

    Foundation of Olle Engkvist

    Available from: 2016-08-23 Created: 2016-08-23 Last updated: 2017-11-28Bibliographically approved
    3. Bacteriocin plantaricin NC8 αβ antagonizes Porphyromonas gingivalis infection and induces proliferation of gingival epithelial cells
    Open this publication in new window or tab >>Bacteriocin plantaricin NC8 αβ antagonizes Porphyromonas gingivalis infection and induces proliferation of gingival epithelial cells
    (English)Manuscript (preprint) (Other academic)
    National Category
    Other Basic Medicine
    Research subject
    Biomedicine
    Identifiers
    urn:nbn:se:oru:diva-52858 (URN)
    Available from: 2016-10-06 Created: 2016-10-06 Last updated: 2018-01-14Bibliographically approved
    4. Effects of plantaricin NC8 αβ and antibodies on gingival epithelial cells infected by Porphyromonas gingivalis
    Open this publication in new window or tab >>Effects of plantaricin NC8 αβ and antibodies on gingival epithelial cells infected by Porphyromonas gingivalis
    Show others...
    (English)Manuscript (preprint) (Other academic)
    National Category
    Other Basic Medicine
    Research subject
    Biomedicine
    Identifiers
    urn:nbn:se:oru:diva-52859 (URN)
    Available from: 2016-10-06 Created: 2016-10-06 Last updated: 2018-01-14Bibliographically approved
  • 5.
    Nakka, Sravya Sowdamini
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden. The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden.
    Lönn, Johanna
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden. The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden.
    Starkhammar Johansson, Carin
    Centre for Oral Rehabilitation, Public Dental Health Care, County Council of Östergötland, Linköping, Sweden.
    Bengtsson, Torbjörn
    Örebro University, School of Medicine, Örebro University, Sweden.
    Nayeri, Fariba
    The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden; Division of Infectious Diseases, University Hospital, Linköping, Sweden.
    Antibodies produced in vitro in the detection of periodontal bacteria by using surface plasmon resonance analysis2015In: Clinical and Experimental Dental Research, ISSN 2057-4347, Vol. 1, no 1, p. 32-44Article in journal (Refereed)
    Abstract [en]

    Porphyromonas gingivalis (P. gingivalis) is a major etiological agent associated with periodontitis. This study aims to develop antibodies to P. gingivalis in vitro for real-time detection of bacteria in clinical samples. Lymphocytes were isolated from whole blood of patient treated for periodontitis and were stimulated with P. gingivalis ATCC 33277. B-cell maturation to long-living antibody secreting-plasma cells was studied using flow cytometry and immunofluorescence staining. The antibodies developed in vitro were immobilized onto a CM-5 sensor chip of a biosensor to detect the presence of P. gingivalis in the gingival crevicular fluid of patients with periodontitis compared to periodontally healthy controls (n = 30). Surface plasmon resonance (SPR) analysis was performed to evaluate specific interactions of bacteria in samples with the immobilized antibodies. The results of SPR analysis were compared to the detection of P. gingivalis in the samples using DNA–DNA checkerboard hybridization technique. A clear and distinct change in lymphocyte morphology upon stimulation with P. gingivalis was observed. Anti-P. gingivalis antibodies secreted by CD38+ plasma cells showed the presence of all the four IgG subclasses. The results of DNA–DNA checkerboard analysis were in agreement with that of SPR analysis for the detection of P. gingivalis in patient samples. Furthermore, incubation with anti-P. gingivalis attenuated the bacterial response in SPR. The in vitro method for antibody production developed during this study could be used for an efficient real-time detection of periodontitis, and the attenuating effects of in vitro antibodies suggest their role in passive immunization to prevent periodontitis and their associated risk factors.

  • 6.
    Nakka, Sravya Sowdamini
    et al.
    Örebro University, School of Medical Sciences. The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden.
    Palm, Eleonor
    Örebro University, School of Medical Sciences.
    Bengtsson, Torbjörn
    Örebro University, School of Medical Sciences.
    Khalaf, Hazem
    Örebro University, School of Medical Sciences.
    Bacteriocin plantaricin NC8 αβ antagonizes Porphyromonas gingivalis infection and induces proliferation of gingival epithelial cellsManuscript (preprint) (Other academic)
  • 7.
    Nakka, Sravya Sowdamini
    et al.
    Örebro University, School of Medical Sciences. The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden.
    Palm, Eleonor
    Örebro University, School of Medical Sciences.
    Nayeri, Fariba
    The Institution for Protein Environmental Affinity Surveys, PEAS Institut AB, Linköping, Sweden; Division of Infectious Diseases, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden.
    Bengtsson, Torbjörn
    Örebro University, School of Medical Sciences.
    Khalaf, Hazem
    Örebro University, School of Medical Sciences.
    Effects of plantaricin NC8 αβ and antibodies on gingival epithelial cells infected by Porphyromonas gingivalisManuscript (preprint) (Other academic)
  • 8.
    Sorour, Ashraf E.
    et al.
    Department of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University, Cairo, Egypt.
    Lönn, Johanna
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Nakka, Sravya Sowdamini
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Nayeri, Tayeb
    The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.
    Nayeri, Fariba
    Division of Infectious Diseases, Department of Medical and Health Sciences, University Hospital, Linköping, Sweden; The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.
    Evaluation of hepatocyte growth factor as a local acute phase response marker in the bowel: the clinical impact of a rapid diagnostic test for immediate identification of acute bowel inflammation2015In: Cytokine, ISSN 1043-4666, E-ISSN 1096-0023, Vol. 71, no 1, p. 8-15Article in journal (Refereed)
    Abstract [en]

    Background: There are no rapid tests that can distinguish contagious gastroenteritis, which requires isolation at its onset, from exacerbation of chronic inflammatory bowel disease (IBD) or bowel engagement in the course of systemic inflammatory response syndrome (SIRS). Hepatocyte growth factor (HGF) is an acute phase cytokine that is produced at the site of injury. It has high affinity to sulfated glycan, and this binding affinity is lost during chronic inflammation. The fecal pH strongly impacts the prognosis for severe bowel disease. We developed a strip test to evaluate HGF as a local acute phase response marker in the bowel. This test assessed the binding affinity of HGF to sulfated glycans in fecal samples and determined fecal pH as an indicator of illness severity.

    Methods: Fresh feces from patients with diarrhea (n = 513) were collected and tested blindly, and information about patient illness course and outcome was collected. Patients were classified based on the focus of inflammation and the cause of the symptoms. Objectively verified diagnoses of infectious gastroenteritis (n = 131) and IBD onset/exacerbation and bowel cancer (n = 44) were used to estimate the performance of the test strip. ELISA was performed on 101 freeze-thawed feces samples to determine the fecal HGF levels.

    Results: The test rapidly distinguished infectious gastroenteritis from non-infectious inflammatory causes of diarrhea (sensitivity, 87.96%; specificity, 90.9%; positive predictive value, 96.6%; negative predictive value, 71.4%; accuracy, 89.1%). Fecal pH (p < 0.0001) and mortality within 28 days of sampling (p < 0.04) was higher in patients with sepsis/SIRS and diarrhea. The concentration of HGF was higher in strip test-positive stool samples (p < 0.01).

    Conclusions: HGF is a good local acute phase response marker of acute bowel inflammation. Test-strip determination of the binding affinity of fecal HGF to sulfated glycan was a rapid, equipment-free way to assess patients with diarrhea and to guide the diagnostic and therapeutic approaches on admission. (C) 2014 The Authors. Published by Elsevier Ltd.

  • 9.
    Wu, Chongcong
    et al.
    The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden; Maternal and Children Health Care Hospital of Zhuhai City, Zhuhai, China .
    Nakka, Sravya
    Örebro University, School of Medical Sciences. The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden .
    Mansouri, Sepahdar
    The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden .
    Bengtsson, Torbjörn
    Örebro University, School of Medical Sciences.
    Nayeri, Tayeb
    The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden .
    Nayeri, Fariba
    The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden; Division of Infectious Diseases, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden .
    In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments2016In: BMC Microbiology, ISSN 1471-2180, E-ISSN 1471-2180, Vol. 16, no 1, article id 209Article in journal (Refereed)
    Abstract [en]

    Background: There is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the host’s defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms. The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms.

    Methods: Lymphocytes were separated from whole blood of patients (n = 10) and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system (SPR) and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies.

    Results: The produced antibodies recognized bacteria with high sensitivity (SPR). The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy.

    Conclusion: In the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.

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