Production of interferon‐gamma, interleukin‐6, and interleukin‐1β by human peripheral blood mononuclear cells stimulated with novel lys‐gingipain synthetic peptides

Abstract

Background: Periodontitis is a progressive inflammatory process, and its pathogenesis is related to the presence of a dysbiotic subgingival biofilm that elicits the immune response. Porphyromonas gingivalis is a keystone pathogen, and its Lys-gingipain (Kgp) virulence factor is involved in the pathogen-host interaction through the production of cytokines by host cells, but the specific mechanisms of this interaction have not been elucidated. The present study evaluated the in vitro production of interferongamma (IFN-), interleukin (IL)-6, and IL-1 cytokines in response to antigenic stimulation of peripheral blood mononuclear cells (PBMCs) with novel Kgp synthetic peptides. Methods:Our previous in silico study predicted 16 immunogenic peptides from Kgp protein. Nine peptides derived from different regions of the protein were chemically synthesized. The synthetic peptides Kgp12, 17, and 18 were selected based on the immunoglobulin G immunoreactivity in the serum of patients with periodontitis (P) and individuals without periodontitis (WP), and they were used in in vitro stimulation of PBMC derived from groups P and WP. Enzyme-linked immunosorbent assay and microsphere-based flow cytometric assay were used to verify the levels of the cytokines produced in PBMC cultures after 48 hours.Results: Kgp12, 17, and 18 peptides induced lower production of IFN-. Kgp12 induced higher levels of IFN-in WP than in P individuals. Kgp12 induced higher production of IL-6 and IL-1 compared with the other stimuli. Conclusion:The novel Kgp synthetic peptides tested herein are immunogenic peptides (epitopes) since they induced the production of cytokines by PBMC and therefore may be useful tools in evaluating the pathogen-host interaction. K E Y W O R D Scytokines, immunology, microbiology, periodontitis J Periodontol. 2019;90:993-1001.

Heme Competition Triggers an Increase in the Pathogenic Potential of Porphyromonas gingivalis in Porphyromonas gingivalis-Candida albicans Mixed Biofilm

“…Participating in the utilization of heme, the gingipains are also one of the major secreted proteins of P. gingivalis and are an important virulence factor (de Diego et al, 2014;Santos-Lima et al, 2019). Imamura et al (1994) reported that HRgpA and RgpB can accelerate the formation of gingival crevicular fluid (GCF) and the progression of inflammation.…”

The cytokine network involved in the host immune response to periodontitis

“…83,84 Because of its pathogen-related upstream regulation, broad derivation from multiple cell groups and direct effect on immune response and osteoclastic activity, IL-6 was applied widely as a representative pro-inflammatory and damage-related cytokines in in vitro studies and in vivo animal studies and shown to be upregulated after stimulation and downregulated after treatment. [85][86][87][88][89] TNF family TNF was first recognized as a tumour necrotizing substance and named in 1975. 90 The two TNF family members, TNF-α and TNF-β, were purified and identified in the 1980s and share the same membrane receptor.…”

Conserved immunogenic peptides of Ebola glycoprotein elicit immune response in human peripheral blood mononuclear cells

“…32 In another study, cytokine measurement was carried out with the help of ELISA in PBMC stimulated with lysgingipain synthetic peptides belonging to Porphyromonas gingivalis. 33 In the present work, P3 emerged as the most potent candidate, as it was able to induce IFN-γ secretion in all (10) the blood samples followed by P2 ( 7) and P1 (6). Moreover, significant positive correlation was observed between peptide specific cell proliferation and IFN-γ production which jointly represents the potential of these peptides to elicit immune response.…”