Immunoassay standardization for the detection of immunoglobulin A (IgA) against Porphyromonas gingivalis antigens in saliva of individuals with and without leprosy.

Leprosy reactions are immune processes that cause neural damage in individuals with leprosy. As periodontitis is an infectious disease related to its development, specific antibodies to periodontal pathogens must be evaluated to better understand the humoral mechanisms underlying this relationship. Therefore, the objective of this study was to standardize an immunoassay to measure IgA specific to P. gingivalis antigens in the saliva of individuals with leprosy. An ELISA checkerboard titration was performed. A validation test involving 53 individuals with leprosy, 24 with and 19 without periodontitis, was conducted and a ROC curve constructed to calculate sensitivity and specificity. The coefficient of the optical densities was 2.21 and 2.66 for P. gingivalis crude extract and the recombinant protein HmuY, respectively. Sensitivity and specificity for the P. gingivalis crude extract were 66.7% and 73.7%, respectively, and for HmuY, were 62.5% and 52.6%, respectively. Specific recognition of P. gingivalis occurred predominantly in individuals with periodontitis, which validates the use of this test for studying periodontitis in individuals with leprosy.Trial registration CAEE 64476117.3.0000.0049, 21/07/2017, retrospectively registered.

In silico analysis as a strategy to identify candidate epitopes with human IgG reactivity to study Porphyromonas gingivalis virulence factors.

Porphyromonas gingivalis (Pg) is one of the main pathogens in chronic periodontitis (CP). Studies on the immunogenicity of its virulence factors may contribute to understanding the host response to infection. The present study aimed to use in silico analysis as a tool to identify epitopes from Lys-gingipain (Kgp) and neuraminidase virulence factors of the Pg ATCC 33277 strain. Protein sequences were obtained from the NCBI Protein Database and they were scanned for amino acid patterns indicative of MHC II binding using the MHC-II Binding Predictions tool from the Immune Epitope Database (IEDB). Peptides from different regions of the proteins were chemically synthesized and tested by the indirect ELISA method to verify IgG immunoreactivity in serum of subjects with CP and without periodontitis (WP). T cell epitope prediction resulted in 16 peptide sequences from Kgp and 18 peptide sequences from neuraminidase. All tested Kgp peptides exhibited IgG immunoreactivity whereas tested neuraminidase peptides presented low IgG immunoreactivity. Thus, the IgG reactivity to Kgp protein could be reaffirmed and the low IgG reactivity to Pg neuraminidase could be suggested. The novel peptide epitopes from Pg were useful to evaluate its immunoreactivity based on the IgG-mediated host response. In silico analysis was useful for preselecting epitopes for immune response studies in CP.

Porphyromonas gingivalis HmuY-induced production of interleukin-6 and IL-6 polymorphism in chronic periodontitis.

BACKGROUND: In chronic periodontitis (CP), the gene polymorphism of interleukin-6 (IL-6) to 174C/G has been associated with the altered production of this cytokine. The aim of this pilot study is to compare the allelic and genotypic frequencies in patients with CP with control individuals without periodontitis (NP) and to measure the production of IL-6 by whole blood cells stimulated with Porphyromonas gingivalis HmuY protein. METHODS: DNA was isolated from peripheral blood cells of 49 patients with CP and 60 control individuals classified as NP, and genotyping was performed by polymerase chain reaction using sequence-specific primers. Whole blood cells from 29 patients with CP and 30 control individuals were stimulated for 48 hours with HmuY, and IL-6 levels were measured using enzyme-linked immunosorbent assay. RESULTS: The proportion of individuals carrying the G allele at position -174 of the IL-6 gene was higher in the group with CP (85.7%) than in the normal control group (73.3%; P <0.03). P. gingivalis HmuY-induced production of IL-6 was higher in the group with CP (P <0.05). CONCLUSIONS: Our findings suggest that P. gingivalis HmuY may be associated with increased IL-6 production during CP. Furthermore, patients with periodontitis and individuals with higher HmuY-induced production of IL-6 show a high frequency of the G allele at position -174.

Oral health profile in patients infected with HTLV-1: clinical findings, proviral load, and molecular analysis from HTLV-1 in saliva.

Human T-lymphotropic virus type 1 (HTLV-1) is associated with adult T-cell leukemia (ATL) and HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) and has also been implicated in several disorders, including periodontal disease. The proviral load is an important biological marker for understanding HTLV-1 pathogenesis and elucidating whether or not the virus is related to the clinical manifestation of the disease. This study describes the oral health profile of HTLV-1 carriers and HAM/TSP patients in order to investigate the association between the proviral load in saliva and the severity of the periodontal disease and to examine virus intra-host variations from peripheral blood mononuclear cells and saliva cells. It is a cross-sectional analytical study of 90 individuals carried out from November 2006 to May 2008. Of the patients, 60 were HTLV-1 positive and 30 were negative. Individuals from the HTLV-1 positive and negative groups had similar mean age and social-economic status. Data were analyzed using two available statistical software packages, STATA 8.0 and SPSS 11.0 to conduct frequency analysis. Differences of P < 0.05 were considered statistically significant. HTLV-1 patients had poorer oral health status when compared to seronegative individuals. A weak positive correlation between blood and saliva proviral loads was observed. The mean values of proviral load in blood and saliva in patients with HAM/TSP was greater than those in HTLV-1 carriers. The HTLV-1 molecular analysis from PBMC and saliva specimens suggests that HTLV-1 in saliva is due to lymphocyte infiltration from peripheral blood. A direct relationship between the proviral load in saliva and oral manifestations was observed.

Tratamento para o controle da infecção pelo virus HTLV-1 e a saude bucal dos pacientes / Treatment for the control of HTLV-1 virus infection and the oral health of patients

Introdução: O virus linfotropico humano de celulas T do tipo 1 (HTLV-1) pertence a familia Retroviridae e a subfamilia Oncornavirus. A infecção pode permanecer por decadas sem manifestação clinica. No Brasil, a Bahia apresenta o mais elevado indice de prevalência da infecção. Manifestações clinicas associadas a infecção pelo virus incluem a PET/MAH, LLTA e uveite. A doença ‚ controlada por tratamento multidisciplinar. Antiinflamatórios, antidepressivos, antibi¢ticos e hormonios são empregados. Al‚m da possibilidade de desenvolver a Sindrome de Sjãgren, que afeta a sa£de bucal, os medicamentos utilizados por estes pacientes podem apresentar efeitos colaterais modificando a salivação e/ou causando lesães na mucosa oral. Objetivo: Descrever os medicamentos mais utilizados no controle da infecção pelo HTLV-1 e sua relação com a saude bucal para fornecer aos cirurgiães-dentistas da rede publica e privada informa‡ães relevantes para o atendimento odontologico a estes pacientes. Método: Foi realizado levantamento bibliogr fico em literatura especializada e os medicamentos foram relacionados quanto a categoria farmacológica, mecanismo de ação e influencia sobre a saude bucal de acordo com as informações apresentadas pelos fabricantes. Resultados: Excetuando-se o Danazol, a Pentoxifilina, a Fosfomicina e a Nitrofurantoina os demais medicamentos podem causar danos a cavidade oral. Os antiinflamatórios podem provocar lesão herpatica. Os antibióticos e antidepressivos, na grande maioria, causam boca seca. Zalcitabina e Carbamazepina podem causar ulceração bucal. O antidepressivo Imipramina além de causar boca seca e problemas gengivais pode também, aumentar os efeitos pressores do vasoconstrictor utilizado em anestesia oral. Conclusão: Os cirurgiões dentistas, particularmente em areas endemicas para o HTLV-1, devem conhecer os efeitos colaterais dos medicamentos utilizados no controle da infecção para garantir um atendimento odontológico de qualidade a este grupo especial de pacientes.

Introduction: The human T-lymphotropic virus Type I (HTLV-1) belongs to the Retroviridae family and Oncornavirus subfamily. Infection may remain for decades without clinical manifestation. In Brazil, the state of Bahia presents the highest prevalence of this infection. Clinical manifestations associated with HTLV-1 infection include PET/MAH, LLTA and uveitis. The disease is controlled by multidisciplinary treatment. Anti-Inflammatory drugs, antidepressants, antibiotics and hormones can be used. In addition to the possibility of developing Sjãgren's Syndrome, which affects oral health, the medications used by the patients can produce side effects, modifying the salivary flow and/or causing oral mucosa injuries. Objective: To describe the most frequently used drugs in the control of HTLV-1 infection and their relationship with oral health in order to provide relevant information for the dental treatment of these patients by dentists working at public and private services. Methods: A review of literature was performed and the drugs were listed according to their pharmacological category, mechanisms of action and influence on oral health, based on the information presented by the manufacturers. Results: Except for danazol, pentoxiphiline, phosphomicin and nitrofurantoin, the other drugs may cause oral problems. Anti-inflammatory drugs may cause herpetic lesions. The majority of antibiotics and antidepressants cause dry mouth. Zalcitabin and carbamazepine may cause oral mucosa ulceration. The antidepressant imipramine, in addition to causing dry mouth and gingival problems, may also increase the pressor effects of the vasoconstrictor used in oral anesthesia. Conclusion: Especially in endemic areas for HTLV-1, dentists should be familiar with the side effects of the drugs used in the control of the infection to ensure a high-quality dental treatment for this especial group of patients.