Differential Cytokine Patterns in Mouse Macrophages and Gingival Fibroblasts After Stimulation With Porphyromonas gingivalis or Escherichia coli Lipopolysaccharide

Abstract

Background A major cause of chronic inflammatory periodontal diseases is Porphyromonas gingivalis (P. gingivalis), a non-motile, gram-negative, rod-shaped, anaerobic bacterium. Within gingival tissue, both macrophages and fibroblasts participate in the immune response to foreign entities by releasing cytokines and expressing molecules to recruit and activate lymphocytes. However, the contribution of gingival macrophages and fibroblasts to the immune response to P. gingivalis infection is not fully known. Methods The AMJ2-C8 cell line (AM cells), a mouse alveolar macrophage cell line, and ESK-1 cells, a mouse gingival fibroblast cell line made in our laboratory, were treated with lipopolysaccharide (LPS) from either P. gingivalis or Escherichia coli (E. coli). The expression of immune response molecules was quantified by real-time PCR and enzyme-linked immunoassay. Results AM and ESK-1 cells responded differently to P. gingivalis and E. coli LPS stimulation. The ESK-1 gingival fibroblast cell line was more responsive to E. coli LPS stimulation as seen by elevated levels of interleukin (IL)-6, inducible nitric oxide (iNOS), and monocyte chemotactic protein-1 (MCP-1) expression relative to stimulation by P. gingivalis LPS. Conversely, the AM macrophage cell line was more responsive to P. gingivalis LPS stimulation, particularly for interleukin IL-1β, IL-6, and MCP-1, relative to stimulation by E. coli LPS. Conclusion These findings demonstrate that E. coli LPS induces a stronger cytokine/chemokine response in gingival fibroblasts, while P. gingivalis LPS induces a stronger response in macrophages.

Oral Administration of Prostaglandin E2‐Specific Receptor 4 Antagonist Inhibits Lipopolysaccharide‐Induced Osteoclastogenesis in Rat Periodontal Tissue

“…Additionally, several reports indicated that P. gingivalis LPS showed different and unique patterns from other LPS in inflammatory response 40,49,50 . Additional study using pathogens for specific types of the each disease will help to evaluate clinical significance.…”

Assessing the effect of triethyleneglycol dimethacrylate on tissue repair in 3D organotypic cultures

“…The SV40LT immortalized mouse GF cell line (ESK‐1; gift from Dr. John R Klein) was originally established as primary cells isolated from the gingival tissues (below the mandibular incisors) of adult female 6‐10‐week‐old C57BL/6 mice and infected with the pBABE‐PURO SV40LT retrovirus supernatant. The cells express the fibroblast marker vimentin (Jones, Ekhlassi, Montufar‐Solis, Klein, & Schaefer, ). The SV40LT immortalized mouse DPMC line (iMDP3; gift from Dr. Shuo Chen) was originally established as primary cells, isolated from dental pulps (first molars of mandibles) of 3‐day‐old C57BL/6 mice and transfected with the pSV3neo plasmid containing the coding sequence of SV40LT and neomycin (G418)‐resistance gene (ATCC, no.…”

“…The SV40LT immortalized mouse GF cell line (ESK-1; gift from Dr. John R Klein) was originally established as primary cells isolated from the gingival tissues (below the mandibular incisors) of adult female 6-10-week-old C57BL/6 mice and infected with the pBABE-PURO SV40LT retrovirus supernatant. The cells express the fibroblast marker vimentin (Jones, Ekhlassi, Montufar-Solis, Klein, & Schaefer, 2010).…”

Enhancing the Three-Dimensional Structure of Adherent Gingival Fibroblasts and Spheroids via a Fibrous Protein-Based Hydrogel Cover

“…The culture medium was replaced twice a week with a fresh medium. The ESK-1 cells were obtained as a generous gift from Dr. John R. Klein (University of Texas Health Science Center) [Jones et al, 2010].…”