Interleukin-17 plays a role in exacerbation of inflammation within chronic periapical lesions

Coli M, Vasiliji S, Gazivoda D, Vucevi D, Marjanovi M, Luki A

Diego Halab Robles Programa de Doctorado en Ciencias Mdicas Unversidad Austral de Chile

Backround

Backround

Backround

Development of periapical lesions

Th1 response Bone destruction

IFN- Th2 response Porphyromonas gingivalis IL-4 IL-5 IL-10 humoral response

TFG-

IL-1 TNF-

TFG-
IL-17 IL-6 IL-23 Th-17 Chronic periodontitis

Material and methods

Periapical lesions (n = 38) Tooth extraction (n = 32) Specimens (n = 23) Apical surgery (n = 6)

Symptomatic (n = 12)

Asymptomatic (n = 11)

Gentamycin, 60 g ml -1 Penicillin, 100 U ml -1 Streptomycin, 0.1 mg ml -1 Amphotericin, 100 g ml -1

Preparation of total and mononuclear inflammatory cells

Collagenase type IV DNAse

Remove tissue fragments

Hemocytometer

Centrifugation

Trypan Blue dye exclusion

Suspension

5.0 x 105 - 2.2 x 106 cells per lesion Preparation of PB-MNC; from healthy volunteers (n = 7) and patients with periapical lesions (n = 8)

Cell cultures
RPMI 1640 10% heat-inactivated FCS Phorbol myristate acetate (PMA) Ca2+ ionophore
24 hours

Recombinant human IL-17

Centrifuged and frozen at -70C

Detection of IL-17 and IL-8 in culture supernatants

Quantikine human IL-17 immunoassay IL-8: flow cytometry

Statistical analysis
ANOVA, t-student and r Spearman = 0.05

Monoclonal antibodies and immunocytochemistry

Anti-CD3, -CD4, -CD8, -CD19, and - CD11c Anti-human Ig rabbit conjugated with streptavidin alkaline phosphatase, and Fast Red. 500 cells per lesion

T-type = CD3+

B-type= CD19+/CD38+

Production of IL-17 by PL-MNC in vitro

PMA and Ca2+

% neutrophils and the level of IL-17 in the group of asymptomatic lesions ; r = 0.78; P < 0.01

Conclusions
PL-MNC, isolated from periapical lesions, produce significant amounts of IL-17 upon additional stimulation IL-17, by stimulating the production of IL-8, plays a role in the exacerbation of chronic inflammation within chronic periapical lesions.