Cystatin C: immunoregulation role in macrophages infected with Porphyromonas gingivalis

• Published in: PeerJ (San Francisco, CA) Vol. 12; p. e17252
• Main Authors: Blancas-Luciano, Blanca Esther, Becker-Fauser, Ingeborg, Zamora-Chimal, Jaime, Jiménez-García, Luis, Lara-Martínez, Reyna, Pérez-Torres, Armando, González Del Pliego, Margarita, Aguirre-Benítez, Elsa Liliana, Fernández-Presas, Ana María
• Format: Journal Article
• Language: English
• Published: United States PeerJ. Ltd 30-04-2024; PeerJ Inc
• Subjects: Analysis; Apoptosis; Apoptosis - drug effects; Communicable diseases; Cystatin C; Cystatin C - metabolism; Cytokines; Cytokines - metabolism; Enzyme-linked immunosorbent assay; Ethylenediaminetetraacetic acid; Humans; Macrophage; Macrophages; Macrophages - drug effects; Macrophages - immunology; Macrophages - metabolism; Macrophages - microbiology; Metronidazole; Nitric oxide; Nitric Oxide - metabolism; Oral peptide; Periodontitis - drug therapy; Periodontitis - immunology; Periodontitis - microbiology; Periodontitis - pathology; Porphyromonas gingivalis; Porphyromonas gingivalis - immunology; Reactive Oxygen Species - metabolism; Scientific equipment and supplies industry; United States; Germany; Porphyromonas gingivalis; Oral peptide; Cystatin C; Macrophage
• ISSN: 2167-8359; 2167-8359
• DOI: 10.7717/peerj.17252

Periodontitis is a chronic infectious disease, characterized by an exacerbated inflammatory response and a progressive loss of the supporting tissues of the teeth.
is a key etiologic agent in periodontitis. Cystatin C is an antimicrobial salivary peptide that inhibits the growth of
. This study aimed to evaluate the antimicrobial activity of this peptide and its effect on cytokine production, nitric oxide (NO) release, reactive oxygen species (ROS) production, and programmed cell death in human macrophages infected with
Monocyte-derived macrophages generated from peripheral blood were infected with
(MOI 1:10) and stimulated with cystatin C (2.75 µg/ml) for 24 h. The intracellular localization of
and cystatin C was determined by immunofluorescence and transmission electron microscopy (TEM). The intracellular antimicrobial activity of cystatin C in macrophages was assessed by counting Colony Forming Units (CFU). ELISA assay was performed to assess inflammatory (TNFα, IL-1β) and anti-inflammatory (IL-10) cytokines. The production of nitrites and ROS was analyzed by Griess reaction and incubation with 2',7'-dichlorodihydrofluorescein diacetate (H
DCFDA), respectively. Programmed cell death was assessed with the TUNEL assay, Annexin-V, and caspase activity was also determined.
Our results showed that cystatin C inhibits the extracellular growth of
. In addition, this peptide is internalized in the infected macrophage, decreases the intracellular bacterial load, and reduces the production of inflammatory cytokines and NO. Interestingly, peptide treatment increased ROS production and substantially decreased bacterial-induced macrophage apoptosis.
Cystatin C has antimicrobial and immuno-regulatory activity in macrophages infected with
These findings highlight the importance of understanding the properties of cystatin C for its possible therapeutic use against oral infections such as periodontitis.