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Application of the In Vitro HoxB8 Model System to Characterize the Contributions of Neutrophil–LPS Interaction to Periodontal Disease

1
Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland
2
Institute of Medical Microbiology and Hygiene, Medical Center–University of Freiburg, Faculty of Medicine, 79104 Freiburg, Germany
3
Malopolska Centre of Biotechnology, Jagiellonian University, 30-387 Krakow, Poland
4
Broegelmann Research Laboratory, Department of Clinical Science, University of Bergen, 5020 Bergen, Norway
5
Department of Oral Immunity and Infectious Diseases, University of Louisville, School of Dentistry, Louisville, KY 40202, USA
*
Author to whom correspondence should be addressed.
Pathogens 2020, 9(7), 530; https://doi.org/10.3390/pathogens9070530
Received: 5 June 2020 / Revised: 21 June 2020 / Accepted: 29 June 2020 / Published: 1 July 2020
(1) Background: Studying neutrophils in vitro is difficult since these cells are terminally differentiated and are easily activated during isolation. At the same time, most of the available model cell lines are associated with certain limitations, such as functional deficiency or a lack of expression of surface markers characteristic of neutrophils. P. gingivalis is a periodontopathogen that causes dysbiosis in subgingival bacterial biofilm. This triggers the accumulation of functional neutrophils in the periodontium. However, until now, the specific effects of P. gingivalis-derived lipopolysaccharide on neutrophil functions have not been analyzed. (2) Methods: The impact of two variants of commercially available P. gingivalis endotoxin on neutrophil functions was tested using the HoxB8 in vitro system that is well suited to analyze neutrophil response to different stimuli in a controlled manner. (3) Results: The Standard P. gingivalis lipopolysaccharide (LPS), known to activate cells through Toll-like receptor 2 (TLR2)- and Toll-like receptor 4 (TLR4)-dependent pathways, prolonged neutrophil survival and exhibited pro-inflammatory effects. In contrast, Ultrapure LPS, binding exclusively to TLR4, neither protected neutrophils from apoptosis, nor induced an inflammatory response. (4) Conclusion: Two variants of P. gingivalis-derived LPS elicited effects on neutrophils and, based on the obtained results, we concluded that the engagement of both TLR2 and TLR4 is required for the manipulation of survival and the stimulation of immune responses of HoxB8 neutrophils. View Full-Text
Keywords: Porphyromonas gingivalis; innate immune system; virulence factors; apoptosis; neutrophil biology Porphyromonas gingivalis; innate immune system; virulence factors; apoptosis; neutrophil biology
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Sochalska, M.; Stańczyk, M.B.; Użarowska, M.; Zubrzycka, N.; Kirschnek, S.; Grabiec, A.M.; Kantyka, T.; Potempa, J. Application of the In Vitro HoxB8 Model System to Characterize the Contributions of Neutrophil–LPS Interaction to Periodontal Disease. Pathogens 2020, 9, 530.

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