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1.
PLoS One ; 14(6): e0218319, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31211815

RESUMO

OBJECTIVES: Primary Sjögren's syndrome (pSS) is an autoimmune disease characterized by reduced lacrimal and salivary secretion. Sicca symptoms together with fatigue and musculoskeletal pain can significantly reduce the patients' quality of life. Furthermore, low salivary secretion may disrupt the oral microbial homeostasis. The aim of this study was to compare the salivary microbiota from pSS patients with patients with sicca symptoms not fulfilling the classification criteria for pSS (non-SS), and with healthy controls without sicca complaints. METHODS: Pellets from centrifuged chewing-stimulated whole saliva from pSS patients (n = 15), non-SS sicca patients (n = 15) and healthy controls (n = 15) were prepared. DNA was extracted and analyzed by 16S rRNA gene sequencing. The acquired sequencing data were performed using the human oral microbiome database (HOMD). RESULTS: We detected 42, 45, and 34 bacterial genera in saliva samples from pSS patients, non-SS sicca patients, and healthy controls, respectively. The most abundant genera in all samples were Prevotella, Veillonella, Streptococcus, and Haemophilus. At species level Streptococcus intermedius, Prevotella intermedia, Fusobacterium nucleatum subsp. vincentii, Porphyromonas endodontalis, Prevotella nancensis, Tannerella spp., and Treponema spp. were detected in the samples from pSS and non-SS only, while Porphyromonas pasteri was mostly found among the healthy controls. CONCLUSION: Our study indicated dysbiosis in the salivary microbiota from pSS and non-SS patients compared to healthy controls. Additionally, the results showed that the salivary microbiome in the pSS group differed significantly from the non-SS group.


Assuntos
Disbiose/microbiologia , Microbiota/genética , Dor Musculoesquelética/microbiologia , Síndrome de Sjogren/microbiologia , Bactérias/classificação , Bactérias/genética , Disbiose/metabolismo , Disbiose/patologia , Feminino , Haemophilus/genética , Haemophilus/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Boca/microbiologia , Dor Musculoesquelética/complicações , Dor Musculoesquelética/genética , Dor Musculoesquelética/fisiopatologia , Prevotella/genética , Prevotella/isolamento & purificação , Qualidade de Vida , RNA Ribossômico 16S/genética , Saliva/microbiologia , Síndrome de Sjogren/complicações , Síndrome de Sjogren/genética , Síndrome de Sjogren/fisiopatologia , Streptococcus/genética , Streptococcus/isolamento & purificação , Veillonella/genética , Veillonella/isolamento & purificação
2.
Int J Oral Maxillofac Surg ; 45(3): 318-22, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26554824

RESUMO

Temporomandibular joint (TMJ) involvement in juvenile idiopathic arthritis (JIA) occurs in up to 80% of affected children. The purpose of this study was to investigate the presence of bacterial DNA in synovial fluid, and to compare this with clinical and immunological findings in children with JIA, adults with persistent JIA, and adults with rheumatoid arthritis, in order to detect whether bacteria contribute to inflammation in TMJ arthritis. Synovial fluid and skin swab samples were collected from 30 patients (54 TMJs). Bacterial detection was performed using 16S rRNA pyrosequencing. Bacterial DNA was detected in 31 TMJs (57%) in 19 patients (63%). A positive statistically significant correlation was registered between bacterial DNA detected in TMJ synovial fluid and the following factors: total protein concentration in synovial fluid, interleukin 1ß, tumour necrosis factor alpha, adrenocorticotropic hormone, and adiponectin, as well as the duration of the general medical disease. Fourteen different bacterial species were detected in synovial fluid. Bacterial DNA in TMJ synovial fluid without contamination was detected in more than 50% of the patients. Studies are needed to evaluate the consequences of this bacterial DNA in synovial fluid with regard to TMJ arthritis.


Assuntos
Artrite Juvenil/microbiologia , Artrite Reumatoide/microbiologia , Líquido Sinovial/microbiologia , Articulação Temporomandibular/microbiologia , Adolescente , Adulto , Idoso de 80 Anos ou mais , Artrite Juvenil/imunologia , Artrite Reumatoide/imunologia , Criança , DNA Bacteriano/análise , Feminino , Humanos , Inflamação , Masculino , Reação em Cadeia da Polimerase , Líquido Sinovial/imunologia
3.
Scand J Immunol ; 79(5): 338-45, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24612355

RESUMO

Chronic periodontitis (CP) and atherosclerotic and aortic aneurysmal vascular diseases (VD) are inflammatory conditions that share a number of predisposing factors. They have a complex genetic heritability and may share genetic risk factors, but a well-defined relationship is still not determined. In addition, distinct genetic patterns of predisposition have been associated with these diseases. Here, we investigated the association of polymorphisms in the IL-1 gene locus with CP in a case-case study analysing VD patients with or without CP. Seventy-four patients with VD of whom 36 had CP were genotyped for single nucleotide polymorphisms in the IL1A -889 (rs1800587), IL1B +3954 (rs1143634) and IL1B at -511 (rs16944) genes and for VNTR polymorphisms in the IL1RN gene. A significantly higher frequency (17%) for allele 1 (four repeats) of the IL1RN VNTR gene was found among the VD patients with CP compared to those without CP. In addition, the frequency of the IL1RN VNTR genotypes 1/1 (4/4 repeats) and 2/2 (2/2 repeats) were significantly higher and lower, respectively, in VD patients with CP. These findings suggest an association of genetic polymorphisms in the IL1-gene locus with risk for CP in patients with VD. The carriage of the risk genotypes, the development and the subsequent influence of CP on systemic health may constitute an additional burden in the pathogenesis of VD. This emphasizes the importance of effective periodontal treatment in patients with VD.


Assuntos
Aneurisma Aórtico/epidemiologia , Aterosclerose/epidemiologia , Periodontite Crônica/epidemiologia , Predisposição Genética para Doença , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-1/genética , Idoso , Idoso de 80 Anos ou mais , Aneurisma Aórtico/genética , Aterosclerose/genética , Periodontite Crônica/genética , Análise Mutacional de DNA , Feminino , Frequência do Gene , Estudos de Associação Genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Países Escandinavos e Nórdicos
4.
J Clin Virol ; 53(4): 364-6, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22261124

RESUMO

BACKGROUND: Human papillomaviruses (HPV) are involved in the etiology of cervix cancer, but it is still unclear whether they play a role in related oral lesions. OBJECTIVES: The presence of HPV in oral leukoplakia biopsies (n=50) and oral squamous carcinoma biopsies (n=50) was compared to normal oral mucosa swabs (n=50) for the purpose of indicating a possible etiological role for the virus. STUDY DESIGN: DNA was extracted from tissue biopsies and from mucosa swabs of control samples. Nested PCR was performed with primers targeting conserved sequences within the capsid gene L1. PCR products were sequenced to identify the HPV genotype. RESULT: The results reveal a profile of low-risk HPV genotypes in oral leukoplakia similar to that in healthy controls, while HPV was less frequently observed in oral squamous carcinoma. CONCLUSIONS: HPV does not seem to represent an important causal factor for the development of oral leukoplakia or oral squamous carcinoma.


Assuntos
Carcinoma de Células Escamosas/virologia , Leucoplasia Oral/virologia , Mucosa Bucal/virologia , Neoplasias Bucais/virologia , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Carcinoma de Células Escamosas/patologia , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/patologia , Papillomaviridae/classificação , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA , Adulto Jovem
5.
Virus Genes ; 23(2): 215-26, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11724277

RESUMO

Cytomegalovirus infection is a major cause of morbidity in immunocompromised patients. The major immediate early promoter/enhancer (MIEP) of the human cytomegalovirus controls the expression of the immediate early genes 1 and 2 which play a central role both in primary and reactivated human cytomegalovirus (HCMV)-infections. Our previous studies have shown that co-infection of A549 cells with human cytomegalovirus and human polyomavirus BK resulted in enhanced expression of the immediate early genes 1 and 2 and that the early gene products of BK virus trans-activated the MIEP. However, neither the MIEP sequences required for mediating this trans-activation, nor the contribution of the individual BK virus early gene products were examined. The MIEP contains multiple binding sites for the transcription factors CREB, AP1, Sp1 and NFkappaB, which may mediate polyomavirus large T- or small t-antigens-induced promoter activation. Transient transfection studies in A549 cells demonstrated that SV40 large T-antigen, but not small t-antigen, trans-activated MIEP activity approximately 9-fold. Mutations in individual binding motifs in the context of the complete MIEP did not impair traits-activation by large T-antigen. The level of induction of a truncated MIEP consisting of a single set of CRE/AP1, NFkappaB, and Sp1 binding motifs by large T-antigen was reduced 2-fold compared to the full length MIEP. Extended truncations diminished trans-activation by large T-antigen. To determine the contribution of a single binding motif in the trans-activation by large T-antigen, a CRE/AP1, an NFkappaB, an Sp1, or a non-consensus Sp1-motif, respectively, was linked to the MIEP TATA-sequence respecting the natural spacing between the two transcription regulatory elements. Only the MIEP TATA-box with the correctly spaced non-consensus Sp1 binding site (GT-motif) was stimulated by large T-antigen. These results suggest that an isolated non-consensus Sp1-motif is important for trans-activation of the MIEP by large T-antigen, but that other cis-acting elements can compensate for this element in the context of the whole MIEP.


Assuntos
Antígenos Transformantes de Poliomavirus/fisiologia , Genes Precoces , Regiões Promotoras Genéticas , Ativação Transcricional/fisiologia , Antígenos Transformantes de Poliomavirus/genética , Antígenos Transformantes de Poliomavirus/metabolismo , Sequência de Bases , Humanos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fatores de Transcrição/metabolismo , Células Tumorais Cultivadas
6.
APMIS ; 106(2): 305-14, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9531964

RESUMO

The interaction of herpes simplex virus (HSV) with mononuclear phagocytes (MP), i.e. monocytes and macrophages, is of importance for the pathogenesis of HSV infections. MP are known to play a significant role in the cellular defence against infections with HSV, but it has also been shown that HSV-1 affects MP. The infection of these cells at different stages of differentiation has different outcomes, and may result in the alteration of important cellular functions. HSV-1 inhibits the morphological differentiation of human monocytes, and this inhibition occurs in spite of the fact that human monocytes are non-permissive to HSV-1. We have studied the effect of HSV infection of monocytes and macrophages on production of essential cytokines and related this effect to the reproduction of the virus. Blood-derived MP were cultured in vitro and inoculated with HSV at different stages of differentiation. Replication of the virus was measured by infectivity titration, detection of HSV antigens by immunofluorescence and detection of HSV-specific mRNA. In monocytes, no viral replication and no production of late protein was seen. HSV IE gene was transcribed in monocytes from some donors, but not from others. In macrophages, virus replicated, but less efficiently than in fully permissive fibroblast cells. The production of IL-1 beta, IL-6 and TNF-alpha in both non-permissive monocytes and permissive macrophages was assayed both at the transcriptional level, as mRNA, and as protein released from the cells. Production of cytokines by MP was affected by HSV-1. The level of cytokine mRNA and cytokine protein did not correspond for all cytokines, which may suggest that translational regulation and/or cytokine inhibitors are important in the regulation of the cytokine response. The cytokine modulation, both at the transcriptional level and measured as biological activity, was different in monocytes and macrophages, and varied between different donors. Our results indicate a relation between permissiveness and cytokine response in mononuclear phagocytes infected with HSV-1. Such a relation may be of importance to both intrinsic and extrinsic defence mechanisms of MP against HSV-1. Our study also demonstrates that even the functions of non-permissive cells such as blood-derived monocytes may be affected by viral infections.


Assuntos
Herpesvirus Humano 1/fisiologia , Macrófagos/fisiologia , Macrófagos/virologia , Monócitos/fisiologia , Monócitos/virologia , Diferenciação Celular , Células Cultivadas , Citocinas/genética , Herpes Simples/virologia , Humanos , Macrófagos/citologia , Monócitos/citologia , RNA Mensageiro/análise , Replicação Viral
7.
APMIS ; 106(12): 1194-203, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10052729

RESUMO

We have studied the effect of herpes simplex virus type 1 (HSV1) infection on in vitro differentiation of blood-derived human monocytes into macrophages using morphological, functional and biochemical parameters that alter during macrophage differentiation. Purified preparations of HSV modified the monocyte-macrophage differentiation, in spite of the fact that the virus did not replicate in monocytes. Disappearance of expression of a monocyte-specific surface antigen and the typical development of morphological appearance were delayed in HSV- infected cells. Production of the lysosomal enzyme acid phosphatase, which normally increases during differentiation, was also reduced in infected cells. Transcription of the oncogenes c-myc and c-fos, and the Hsp70 gene was modified in cells from some donors but not in other cell preparations. Possible mechanisms of these effects are discussed.


Assuntos
Herpesvirus Humano 1/fisiologia , Monócitos/fisiologia , Fosfatase Ácida/sangue , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Genes fos , Proteínas de Choque Térmico HSP70/genética , Humanos , Contagem de Leucócitos , Transcrição Gênica
8.
APMIS ; 105(5): 402-9, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9201242

RESUMO

We have investigated the effect of adenovirus 2 (Ad2) infection on human monocytes and monocyte-derived macrophages with regard to expression of TNF-alpha and IL-1 beta. In monocytes, the virus was bound to the surface without being internalized. On the other hand, Ad2 was internalized by macrophages. No virus replication and no transcription of the Ad2 early genes was observed in either of the cells. Ad2 infection induced transient increase in the mRNA levels for TNF-alpha and IL-1 beta in both monocytes and in macrophages, although the kinetics of the transcription was slightly different. The production of both cytokines, measured by ELISA tests, was enhanced in monocytes. In macrophages, a slight enhancement of TNF-alpha production was seen, whereas IL-1 beta was not detected. The data indicate that cellular genes might be activated by Ad2 virus infection in nonpermissive cells where no viral gene products could be detected.


Assuntos
Adenoviridae/fisiologia , Interleucina-1/biossíntese , Macrófagos/metabolismo , Monócitos/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Proteínas E1A de Adenovirus/genética , Proteínas E2 de Adenovirus/genética , Regulação da Expressão Gênica , Humanos , Interleucina-1/genética , Macrófagos/virologia , Monócitos/virologia , RNA Mensageiro , Ativação Transcricional , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa/genética
9.
Virus Res ; 52(1): 61-71, 1997 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9453145

RESUMO

Co-infections or co-habitations of cells by two or more viruses may occur in the human organism. Human cytomegalovirus (HCMV) and the human polyomavirus BK (BKV) have common host cells and may both establish lifelong latency/persistence following primary infection. Both viruses may become reactivated by immunosuppression or other conditions which upset host-virus balance, and they encode gene products with the inherent potential of acting as heterologous transacting factors for expression of cellular or viral genes. It has been shown that HCMV induces gene expression and replication of primate polyomaviruses. We now demonstrate that BKV is able to enhance the expression of HCMV immediate early (IE1 and 2) as well as the early (E) protein pp65 during double infections in semi-permissive cells. By transfection experiments it was established that the phenomenon is due to heterologous transcriptional transactivation of the HCMV major IE promoter (MIEP) by the BKV large T antigen, without contribution from the small t antigen.


Assuntos
Antígenos Virais de Tumores/fisiologia , Vírus BK/imunologia , Citomegalovirus/genética , Regulação Viral da Expressão Gênica , Células Cultivadas , Infecções por Citomegalovirus/genética , Infecções por Citomegalovirus/virologia , Elementos Facilitadores Genéticos , Genes Precoces , Humanos , Regiões Promotoras Genéticas , Ativação Transcricional , Transfecção , Células Tumorais Cultivadas
10.
Blood ; 87(2): 674-81, 1996 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-8555490

RESUMO

In patients with common variable immunodeficiency (CVI), we have previously defined a subgroup of patients (CVIHyper) characterized by decreased numbers of CD4+ lymphocytes in peripheral blood, splenomegaly, and persistent immune activation in vivo, particularly of monocytes/macrophages. To further characterize this hyperactivity, parameters of activation of the tumor necrosis factor (TNF) system (TNF alpha and soluble TNF receptors [sTNFRs]) were measured in 24 patients with CVI and 20 healthy controls. Patients with CVI had significantly higher serum levels of TNF alpha and both types of sTNFRs, with the highest levels in the CVIHyper subgroup. In vitro, peripheral blood mononuclear cells (PBMC) and purified monocytes from CVIHyper patients spontaneously released significantly higher levels, and, after lipopolysaccharide (LPS) stimulation, significantly lower levels of TNF alpha and soluble p75-TNFR than cells from both other CVI patients and healthy controls. CVIHyper patients also had significantly higher TNF alpha:sTNFRs ratios in both serum and in unstimulated PMBC supernatants. The present study demonstrates persistent in vivo activation of the TNF system in CVI, particularly in the CVIHyper subgroup. This activation may contribute to the pathogenesis of both clinical and immunologic manifestations in CVI.


Assuntos
Imunodeficiência de Variável Comum/fisiopatologia , Fator de Necrose Tumoral alfa/fisiologia , Adulto , Idoso , Contagem de Linfócito CD4 , Imunodeficiência de Variável Comum/classificação , Imunodeficiência de Variável Comum/complicações , Imunodeficiência de Variável Comum/imunologia , Feminino , Humanos , Imunofenotipagem , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Pessoa de Meia-Idade , Receptores do Fator de Necrose Tumoral/análise , Esplenomegalia/etiologia
11.
Arch Virol ; 138(1-2): 85-93, 1994.
Artigo em Inglês | MEDLINE | ID: mdl-7980013

RESUMO

During an HCMV infection, transcription of viral and cellular genes are mutually regulated. Several cellular proteins have been implicated in the regulation of the HCMV major immediate early promoter (MIEP) which have been shown to respond to cAMP as well as activation of protein kinase C (PKC). We have examined the effect of an ongoing HCMV infection at the mRNA level for the catalytic and regulatory subunits of protein kinase A (PKA) and alpha and beta isoforms of PKC. There was a moderate elevation for PKA C alpha and RI alpha at immediate early times (0.5-2 h) after HCMV infection. Later in the infection cycle (24-72 h), mRNA level for PKA regulatory subunit RI alpha and PKC alpha were decreased compared with control cells. Messenger RNA levels for the PKA RII alpha and RII beta as well as PKC beta were not affected by HCMV infection. During the infection cycle the PKA subunits and PKC isoforms appeared to be independently regulated. It was also evident that the basal mRNA levels of PKA subunits and the PKC isoforms were sufficient for the PKA and PKC activity required during an HCMV infection in permissive fibroblast cells.


Assuntos
Citomegalovirus/fisiologia , Proteína Quinase C/biossíntese , Linhagem Celular , Subunidade RIIalfa da Proteína Quinase Dependente de AMP Cíclico , Subunidade RIIbeta da Proteína Quinase Dependente de AMP Cíclico , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico/biossíntese , Proteínas Quinases Dependentes de AMP Cíclico/genética , Fibroblastos/enzimologia , Fibroblastos/virologia , Humanos , Proteína Quinase C/genética , RNA Mensageiro/metabolismo
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