In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments.
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.



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.



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.



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.
The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.|Maternal and Children Health Care Hospital of Zhuhai City, Zhuhai, China.
Chongcong
C
Wu
The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.|Department of Medical Sciences, Örebro University, Örebro, Sweden.
Sravya
S
Nakka
The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.
Sepahdar
S
Mansouri
Department of Medical Sciences, Örebro University, Örebro, Sweden.
Torbjörn
T
Bengtsson
The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden.
Tayeb
T
Nayeri
The Institute of Protein Environment Affinity Surveys (PEAS Institut), Linköping, Sweden. fariba.nayeri@regionostergotland.se.|Division of Infectious Diseases, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden. fariba.nayeri@regionostergotland.se.
Fariba
F
Nayeri
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D000917:Antibody Formation / Q000276:immunology*; D001419:Bacteria / Q000187:drug effects / Q000254:growth & development* / Q000276:immunology* / Q000472:pathogenicity; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D001769:Blood / Q000276:immunology; D001826:Body Fluids / Q000276:immunology / Q000382:microbiology; D002908:Chronic Disease; D018920:Coculture Techniques; D060085:Coinfection / Q000175:diagnosis / Q000000981:diagnostic imaging / Q000276:immunology* / Q000382:microbiology*; D019937:Diagnostic Techniques and Procedures; D016983:Enterococcus / Q000276:immunology / Q000472:pathogenicity; D013293:Enterococcus faecalis / Q000276:immunology; D005260:Female; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000188:drug therapy; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D013045:Species Specificity; D020349:Surface Plasmon Resonance / Q000379:methods; D014456:Ulcer / Q000382:microbiology
D000900:Anti-Bacterial Agents
Antibodies; Biofilms; Detection; Electron microscopy; Enterococcus; Lymphocytes; Surface plasmon resonance