Avaliação do fenótipo, perfil imunológico e expressão gênica de portadores da síndrome de Papillon-Lefèvre

Trabalho de Conclusão de Curso (graduação)—Universidade de Brasília, Faculdade de Ciências da Saúde, Departamento de Odontologia, 2019.A síndrome de Papillon-Lefèvre (PL) é uma condição rara que afeta um a quatro indivíduos por milhão. Normalmente é identificada ainda na infância, durante a dentição decídua, devido à perda precoce de dentes, doença periodontal grave e hiperceratose palmo-plantar. Atribui-se à mutação no gene da catepsina C, que afeta os processos imunes e inflamatórios nos portadores da síndrome, o grave quadro periodontal. O objetivo do presente trabalho foi avaliar o fenótipo, o perfil imunológico no plasma sanguíneo de três irmãs portadoras da PL e a expressão de genes, possivelmente relacionados à periodontite, no ligamento periodontal e polpa dentária de uma das irmãs portadoras da PL. A metodologia compreendeu a avaliação das condições clínicas, análises das amostras de polpa dentária e ligamento periodontal por PCR em tempo real (qPCR) e exames sanguíneos. O qPCR revelou alta expressão de IL-3 e TNF-α em polpa e ligamento periodontal do paciente PL, quando comparados a amostra controle não-sindrômica. Os resultados indicaram valores plasmáticos elevados de IgE e IgG nos portadores da síndrome. Portanto, pode-se concluir que as imunoglobulinas e os genes alvos estavam super-expressos no paciente PL, o que pode explicar a resposta inflamatória exacerbada nesses pacientes e o comprometimento estrutural severo dos tecidos periodontais.Papillon-Lefèvre syndrome is a rare condition that affects one to four individuals per million. It is usually identified during childhood during deciduous dentition due to premature tooth loss, severe periodontal disease, and palmoplantar keratoderma. The severe periodontal disease in patients with the syndrome is attributed to the mutation in the cathepsin C gene, which affects their immune and inflammatory processes. The aim of this study was to evaluate the phenotype, the blood plasm immunological profile of the three sisters with PL and the expression of genes possibly related to the periodontitis in the periodontal ligament and dental pulp of one of the three sisters. The methodology comprised: Assessment of their clinical conditions, real-time PCR analysis (qPCR) of the dental pulp and periodontal ligament and blood tests. The qPCR of pulp and periodontal ligament tissues revealed a high expression of IL-3 and TNF-α in the PL patient when compared to the non-syndromic controls. The results showed high concentration of IgE and IgG in the plasm of syndromic patients. Therefore, we can conclude that the immunoglobulins and the target genes were over expressed in PL, which may explain the exacerbated inflammatory response in these patients and the severe structural impairment of the periodontal tissues

Oral Phenotype and Salivary Microbiome of Individuals With Papillon-Lefèvre Syndrome.

Papillon-Lefèvre syndrome (PLS) is an autosomal recessive rare disease, main characteristics of which include palmoplantar hyperkeratosis and premature edentulism due to advanced periodontitis (formerly aggressive periodontitis). This study aimed to characterize the oral phenotype, including salivary parameters, and the salivary microbiome of three PLS sisters, comparatively. Two sisters were toothless (PLSTL1 and PLSTL2), and one sister had most of the teeth in the oral cavity (PLST). Total DNA was extracted from the unstimulated saliva, and the amplicon sequencing of the 16S rRNA gene fragment was performed in an Ion PGM platform. The amplicon sequence variants (ASVs) were obtained using the DADA2 pipeline, and the taxonomy was assigned using the SILVA v.138. The main phenotypic characteristics of PLS were bone loss and premature loss of primary and permanent dentition. The PLST sister presented advanced periodontitis with gingival bleeding and suppuration, corresponding to the advanced periodontitis as a manifestation of systemic disease, stage IV, grade C. All three PLS sisters presented hyposalivation as a possible secondary outcome of the syndrome. Interestingly, PLST salivary microbiota was dominated by the uncultured bacteria Bacterioidales (F0058), Fusobacterium, Treponema, and Sulfophobococcus (Archaea domain). Streptococcus, Haemophilus, and Caldivirga (Archaea) dominated the microbiome of the PLSTL1 sister, while the PLSTL2 had higher abundances of Lactobacillus and Porphyromonas. This study was the first to show a high abundance of organisms belonging to the Archaea domain comprising a core microbiome in human saliva. In conclusion, a PLST individual does have a microbiota different from that of the periodontitis' aggressiveness previously recognized. Due to an ineffective cathepsin C, the impairment of neutrophils probably provided a favorable environment for the PLS microbiome. The interactions of Bacteroidales F0058, Caldivirga, and Sulfophobococcus with the microbial consortium of PLS deserves future investigation. Traditional periodontal therapy is not efficient in PLS patients. Unraveling the PLS microbiome is essential in searching for appropriate treatment and avoiding early tooth loss