RESUMO
The tryptophan (TRP)-kynurenine (KYN) metabolic pathway is a main player of TRP metabolism through which more than 95% of TRP is catabolized. The pathway is activated by acute and chronic immune responses leading to a wide range of illnesses including cancer, immune diseases, neurodegenerative diseases and psychiatric disorders. The presence of positive feedback loops facilitates amplifying the immune responses vice versa. The TRP-KYN pathway synthesizes multifarious metabolites including oxidants, antioxidants, neurotoxins, neuroprotectants and immunomodulators. The immunomodulators are known to facilitate the immune system towards a tolerogenic state, resulting in chronic low-grade inflammation (LGI) that is commonly present in obesity, poor nutrition, exposer to chemicals or allergens, prodromal stage of various illnesses and chronic diseases. KYN, kynurenic acid, xanthurenic acid and cinnabarinic acid are aryl hydrocarbon receptor ligands that serve as immunomodulators. Furthermore, TRP-KYN pathway enzymes are known to be activated by the stress hormone cortisol and inflammatory cytokines, and genotypic variants were observed to contribute to inflammation and thus various diseases. The tryptophan 2,3-dioxygenase, the indoleamine 2,3-dioxygenases and the kynurenine-3-monooxygenase are main enzymes in the pathway. This review article discusses the TRP-KYN pathway with special emphasis on its interaction with the immune system and the tolerogenic shift towards chronic LGI and overviews the major symptoms, pro- and anti-inflammatory cytokines and toxic and protective KYNs to explore the linkage between chronic LGI, KYNs, and major psychiatric disorders, including depressive disorder, bipolar disorder, substance use disorder, post-traumatic stress disorder, schizophrenia and autism spectrum disorder.
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Objectives: Rheumatoid arthritis (RA) is a chronic, inflammatory joint disease with complex pathogenesis involving a variety of immunological events. Recently, it has been suggested that kynurenic acid (KYNA) might be a potential regulator of inflammatory processes in arthritis. KYNA has a definitive anti-inflammatory and immunosuppressive function. The aim of the present study is to investigate the complex effects of a newly synthesized KYNA analog-SZR72 on the in vitro production of tumor necrosis factor-α (TNF-α), tumor necrosis factor-stimulated gene-6 (TSG-6), calprotectin (SA1008/9), SA100 12 (EN-RAGE), and HNP1-3 (defensin-α) in the peripheral blood of patients with RA and the various effects of the disease. Methods: Patients with RA (n = 93) were selected based on the DAS28 score, medication, and their rheumatoid factor (RF) status, respectively. Peripheral blood samples from 93 patients with RA and 50 controls were obtained, and activated by heat-inactivated S. aureus. Parallel samples were pretreated before the activation with the KYNA analog N-(2-N, N-dimethylaminoethyl)-4-oxo-1H-quinoline-2-carboxamide hydrochloride. Following the incubation period (18 h), the supernatants were tested for TNF-α, TSG-6, calprotectin, S100A12, and HNP1-3 content by ELISA. Results: SZR72 inhibited the production of the following inflammatory mediators: TNF-α, calprotectin, S100A12, and HNP1-3 in whole blood cultures. This effect was observed in each group of patients in various phases of the disease. The basic (control) levels of these mediators were higher in the blood of patients than in healthy donors. In contrast, lower TSG-6 levels were detected in patients with RA compared to healthy controls. In addition, the KYNA analog exerted a stimulatory effect on the TSG-6 production ex vivo in human whole blood cultures of patients with RA in various phases of the disease. Conclusion: These data further support the immunomodulatory role of KYNA in RA resulting in anti-inflammatory effects and draw the attention to the importance of the synthesis of the KYNA analog, which might have a future therapeutic potential.
Assuntos
Anti-Inflamatórios/farmacologia , Artrite Reumatoide/imunologia , Mediadores da Inflamação/imunologia , Ácido Cinurênico/análogos & derivados , Idoso , Artrite Reumatoide/sangue , Moléculas de Adesão Celular/sangue , Moléculas de Adesão Celular/imunologia , Feminino , Humanos , Mediadores da Inflamação/sangue , Ácido Cinurênico/farmacologia , Masculino , Pessoa de Meia-Idade , Fator Reumatoide/sangue , Proteínas S100/sangue , Proteínas S100/imunologia , Staphylococcus aureus/imunologia , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/imunologia , alfa-Defensinas/sangue , alfa-Defensinas/imunologiaRESUMO
Purpose: The investigation of anti-inflammatory and immunosuppressive functions of Kynurenic acid (KYNA) is now in focus. There is also substantial evidence that TSG-6 has an anti-inflammatory activity. Therefore, in the present study, we compared the effects of newly synthetized KYNA analogs on the TNF-α production in U-937 monocytic cells in correlation with the effects on the TSG-6 expression. Methods: TNF-α production was measured by ELISA, the TSG-6 expression was determined by RTqPCR method. As cytokine inducers Staphylococcus aureus and Chlamydia pneumoniae were used. Results: KYNA and KYNA analogs attenuated TNF-α production and increased TSG-6 mRNA expression in U-937 cells stimulated by heat inactivated Staphylococcus aureus. In contrast, KYNA and some of the KYNA analogs increased the TNF-α production of C. pneumoniae infected U-937 cells; however, the newly synthetized analogs (SZR104, SZR 105, and SZR 109) exerted significant inhibitory effects on the TNF-α synthesis. The inhibitory and stimulatory effects correlated inversely with the TSG-6 expression. Conclusions: TSG-6 expression following activation with bacterial components could participate in the suppression of inflammatory cytokines, such as TNF-α, We suppose that the elevation of the TSG-6 expression by KYNA and especially by new KYNA analogs might be one of the mechanisms that are responsible for their suppressive effect on TNF-α production as a feedback mechanism. KYNA and KYNA analogs have an important role in influencing TSG-6 expression, and there is a possible benefit of targeting TSG-6 expression by kynurenines in inflammatory conditions following infections.
Assuntos
Moléculas de Adesão Celular/genética , Chlamydophila pneumoniae/imunologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Ácido Cinurênico/farmacologia , Staphylococcus aureus/imunologia , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologia , Moléculas de Adesão Celular/metabolismo , Ensaio de Imunoadsorção Enzimática , Regulação Neoplásica da Expressão Gênica/imunologia , Humanos , Ácido Cinurênico/análogos & derivados , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Monócitos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células U937 , Vacinas Atenuadas/imunologiaRESUMO
BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that involves the selective loss of the upper and lower motor neurons (MNs). Neuroinflammation has been implicated in the pathogenesis of the sporadic form of the disease. We earlier developed immune-mediated animal models of ALS and demonstrated humoral and cellular immune reactions in the nervous system and in the sera of patients and animals. The accumulation of immunoglobulin G (IgG), an elevated intracellular level of calcium, ultrastructural alterations in the MNs, and activation of the microglia were noted in the spinal cord of ALS patients. Similar alterations developed in mice inoculated intraperitoneally with IgG from ALS patients or from an immune-mediated goat model. METHODS: We have now examined whether the intraperitoneal injection of mice with IgG from sporadic ALS patients or from immunized goats with the homogenate of the anterior horn of the bovine spinal cord is associated with changes in the pro-inflammatory (TNF-α and IL-6) and anti-inflammatory (IL-10) cytokines in the spinal cord and serum of the mice. The levels of cytokines were measured by ELISA. RESULTS: Intraperitoneally administered IgG from the ALS patients induced subclinical signs of MN disease, while the injection of IgG from immunized goats resulted in a severe respiratory dysfunction and limb paralysis 24 h after the injections. Significantly increased levels of TNF-α and IL-10 were detected in the spinal cord of the mice injected with the human ALS IgG. The level of IL-6 increased primarily in the serum. The IgG from the immunized goats induced highly significant increases in the levels of all three cytokines in the serum and the spinal cord of mice. CONCLUSIONS: Our earlier experiments had proved that when ALS IgG or IgG from immune-mediated animal models was inoculated into mice, it was taken up in the MNs and had the ability to initiate damage in them. The pathological process was paralleled by microglia recruitment and activation in the spinal cord. The present experiment revealed that these forms of IgG cause significant increases in certain cytokine levels locally in the spinal cord and in the serum of the inoculated mice. These results suggest that IgG directed to the MNs may be an initial element in the damage to the MNs both in human ALS and in its immune-mediated animal models.
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Esclerose Lateral Amiotrófica/sangue , Citocinas/metabolismo , Imunoglobulina G/administração & dosagem , Medula Espinal/metabolismo , Esclerose Lateral Amiotrófica/imunologia , Análise de Variância , Animais , Cálcio/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Cabras , Humanos , Injeções Intraperitoneais , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Masculino , Camundongos , Pessoa de Meia-Idade , Atividade Motora/efeitos dos fármacos , Neurônios Motores/patologia , Neurônios Motores/ultraestrutura , Miastenia Gravis Autoimune Experimental/sangue , Miastenia Gravis Autoimune Experimental/imunologia , Miastenia Gravis Autoimune Experimental/patologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The DEFB4 gene copy numbers were investigated in 206 AD patients and in 250 controls. The levels of the human defensin ß-2 (hBD2) and α-defensins (HNP 1-3) in the sera and in the cerebrospinal fluid (CSF) of the patients and the controls were determined. Higher copy numbers of the DEFB4 gene was observed in AD patients as compared with the controls. The levels of hBD-2 and HNP 1-3 were significantly elevated in the sera and in the CSF of the AD patients These data suggest that both defensin ß-2 and α-defensins have potential role in the development of AD.
Assuntos
Doença de Alzheimer/genética , alfa-Defensinas/genética , beta-Defensinas/genética , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/sangue , Doença de Alzheimer/líquido cefalorraquidiano , Estudos de Casos e Controles , Feminino , Dosagem de Genes , Humanos , Masculino , alfa-Defensinas/sangue , alfa-Defensinas/líquido cefalorraquidiano , beta-Defensinas/sangue , beta-Defensinas/líquido cefalorraquidianoRESUMO
BACKGROUND AND AIMS: Leukoaraiosis (LA), one of the most frequent causes of an age-associated cognitive decline, can be associated with a poor quality of life, leading overall to far-reaching public health problems. Chronic hypoxia of the white matter of the brain may be a factor triggering this entity. LA may develop as a consequence of chronically insufficient cellular energy production and the accumulation of free radicals. METHODS: In this context, after hypothesizing that the number of healthy mitochondria can be crucial in this complex process, a case-control LA study was carried out in which we analyzed the numbers of deleted and non-deleted mitochondria (the common D-loop deletion) per white blood cell. A total of 234 patients with LA and 123 MRI alteration-free subjects served as a control group. RESULTS: Interestingly, it emerged that the ratio of deleted relative to non-deleted mitochondria is strongly associated with the risk of LA. The calculated K ratio in the LA group was significantly lower than the K ratio in the controls (LA: K 0.37 95% CI 0.05; controls: K 0.48, 95% CI 0.076, p < 0.001). CONCLUSIONS: Our study suggests that the ratio of the dmDNA and mDNA can be of great importance in the pathogenesis of LA.
Assuntos
DNA Mitocondrial/genética , Leucoaraiose/patologia , Mitocôndrias/genética , Mitocôndrias/patologia , Idoso , Encéfalo/patologia , Transtornos Cognitivos/etiologia , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Qualidade de VidaRESUMO
AIMS: Recent studies have started to elucidate the contribution of microbiome to the pathogenesis of multiple sclerosis (MS). It is also supposed, that neuropathological alterations might be associated with abnormal expression and regulatory function of antimicrobial peptides (AMPs), including defensins. It is in our interest to investigate the relevance of the single nucleotide polymorphisms (SNPs) of the DEFB1 gene and the copy number polymorphism of the DEFB4 genes in MS. METHODS: DEFBI polymorphisms: c.-20G > A (rsl 1362), DEFB1 c.-44C > G (rsI 800972), DEFB1 c.-52G>A (rsl 799946), and the DEFB4 gene copy number were investigated in 250 MS patients The control patients comprised 232 age- and gender-matched healthy blood donors. The occurrence of the human ß-defensin 2 peptide (hBD2) in the plasma of controls and patients-was determined by ELISA. RESULTS: The DEFB1 c.-44C>G polymorphism the GG protective genotype was much less frequent among patients than among the controls. A higher frequency of a lower (<4) copy number of the DEFB4 gene was observed in the patients with MS as compared with the controls (43% vs. 28%, respectively). The median levels of the circulating hBD2 in the patients were 150.6 +/- 12.71 pg/ml vs. 262.1 +/- 23.82 pg/mI in the control group (p<0.0001). Our results suggest that ß-defensins play role in the development of MS.
Assuntos
Variações do Número de Cópias de DNA , Esclerose Múltipla/genética , Polimorfismo de Nucleotídeo Único , beta-Defensinas/genética , Adulto , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Crônica Progressiva/genética , Esclerose Múltipla Recidivante-Remitente/genéticaRESUMO
AIM: To investigate the genetic background of human defensin expression in type 1 and 2 diabetes. METHODS: Associations between DEFA1/DEFA3 gene copy number polymorphism and diabetes as well as between the promoter polymorphisms of DEFB1 and diabetes were studied. The copy number variation of the DEFA1/DEFA3 genes was determined in 257 diabetic patients (117 patients with type 1 and 140 with type 2 diabetes). The control group consisted of 221 age- and gender-matched healthy blood donors. The cumulative copy numbers of the DEFA1/DEFA3 genes were detected by using quantitative PCR analysis. To evaluate the HNP 1-3 (human neutrophil peptide 1-3 or α-defensin) levels in the circulation, plasma HNP 1-3 concentrations were measured by ELISA. The expression of DEFA1/A3 in peripheral leukocytes of the diabetic patients was measured by quantitative RT PCR analysis. Three SNPs of the human DEFB1 (human defensin ß-1) gene: DEFB1 G-20A (rs11362), DEFB1 C-44G (rs1800972) and DEFB1 G-52A (rs1799946) were genotyped by Custom TaqMan(®) Real Time PCR assay. RESULTS: Significant differences were observed in HNP1-3 levels between the healthy subjects and both groups of diabetic patients. The mean ± SE was 28.78 ± 4.2 ng/mL in type 1 diabetes, and 29.82 ± 5.36 ng/mL in type 2 diabetes, vs 11.94 ± 2.96 ng/mL in controls; P < 0.01 respectively. There was no significant difference between patients with type 1 and type 2 diabetes in the high plasma concentrations of HNP1-3. The highest concentrations of α-defensin were found in diabetic patients with nephropathy (49.4 ± 4.8 ng/mL), neuropathy (38.7 ± 4.8 ng/mL) or cardiovascular complications (45.6 ± 1.45 ng/L). There was no significant difference in the cumulative copy numbers of DEFA1/DEFA3 genes between controls and patients, or between patients with the two types of diabetes. Comparisons of HNP 1-3 plasma level and DEFA1/A3 copy number of the same patient did not reveal significant relationship between defensin-α levels and the gene copy numbers (r (2) = 0.01). Similarly, no positive correlation was observed between the copy numbers and the mRNA expression levels of DEFA1/A3. Regarding the C-44G polymorphism of DEFB1, the GG "protective" genotype was much less frequent (1%-2%) among both groups of patients than among controls (9%). CONCLUSION: Elevated HNP1-3 levels in diabetes are independent of DEFA1/DEFA3 copy numbers, but GG genotype of C-44G SNP in DEFB1 gene may result in decreased defensin ß-1 production.
Assuntos
Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 2/genética , Polimorfismo de Nucleotídeo Único , alfa-Defensinas/genética , beta-Defensinas/genética , Adulto , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Dosagem de Genes , Regulação da Expressão Gênica , Predisposição Genética para Doença , Humanos , Masculino , Fenótipo , Regiões Promotoras Genéticas , RNA Mensageiro/análise , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , alfa-Defensinas/sangue , beta-Defensinas/sangueAssuntos
Calgranulina A/genética , Calgranulina B/genética , Pancreatite/genética , Polimorfismo de Nucleotídeo Único , Proteínas S100/genética , Doença Aguda , Calgranulina A/sangue , Calgranulina B/sangue , Feminino , Frequência do Gene , Genótipo , Humanos , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Pancreatite/sangue , Pancreatite/patologia , Proteínas S100/sangue , Proteína S100A12 , Índice de Gravidade de DoençaRESUMO
In this study we investigated the effects of Candida albicans, Candida krusei, Candida tropicalis and Candida parapsilosis on human beta-defensin 2 (HBD-2) production in Caco-2 intestinal cell line, and the production of alpha-defensins (human neutrophil peptides, HNP 1-3) in peripheral blood. Opportunistic pathogen yeasts can modulate the host immune function by inducing defensins, the natural antimicrobial peptides. Here we show that Candida spp. stimulated HBD-2 expression in and release from Caco-2 cells, with C. albicans inducing the highest levels of HBD-2. Similarly, HNP 1-3 secretion was significantly increased in whole blood after exposure to Candida yeast cells, with C. albicans producing the greatest effect. Our investigations underscore the important role of beta and alpha defensins produced by intestinal epithelial cells locally and neutrophils systemically in the antifungal defense against Candida.
Assuntos
Candida/metabolismo , Defensinas/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Células CACO-2 , Defensinas/genética , Expressão Gênica , Humanos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , alfa-Defensinas/metabolismo , beta-Defensinas/genética , beta-Defensinas/metabolismoRESUMO
Vascular demyelinization of the white matter of the brain is referred to as leukoaraiosis (LA). This very frequent entity is associated with a cognitive decline, thereby resulting in a deteriorating quality of life. Besides poorly controlled hypertension and aging, its development is reported to be associated with an elevated serum homocysteine level. Although the methylenetetrahydrofolate reductase (MTHFR) C677T genetic variant is associated with an elevated serum homocysteine level, it has not been proved to be an independent risk factor for LA. The aim of the present study was to examine whether the MTHFR A1298C genetic variant, which is also believed to be unfavorable, is associated with the presence of LA. The clinical and genetic data on 198 LA patients and 235 neuroimaging alteration-free controls were analyzed. The presence of the A1298C or the 1298CC variant was calculated to be a risk factor for LA, as compared with the absence of both of them. The clustering of the heterozygous A1298C and C677T variants was proved to involve the risk of LA. Our results suggest that the MTHFR A1298C variant confers an independent genetic risk of LA, and this pathological role may be amplified by the MTHFR C677T variant.
Assuntos
Variação Genética/genética , Leucoaraiose/enzimologia , Leucoaraiose/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Mutação Puntual/genética , Adulto , Idoso , Feminino , Triagem de Portadores Genéticos/métodos , Predisposição Genética para Doença/epidemiologia , Predisposição Genética para Doença/genética , Homocisteína/sangue , Homocisteína/genética , Humanos , Hiper-Homocisteinemia/enzimologia , Hiper-Homocisteinemia/epidemiologia , Hiper-Homocisteinemia/genética , Leucoaraiose/epidemiologia , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , Pessoa de Meia-Idade , Família Multigênica , Fatores de RiscoRESUMO
There is developing interest in the role of the kynurenines in the immune function. A considerable amount of evidence has accumulated as concerns interactions between the kynurenine pathway, cytokines and the nervous system. Indoleamine 2,3-dioxygenase (IDO) occupies a key position connecting the immune system and the kynurenine pathway. There are evidences of the immunosuppressive effect of IDO. Following the interferon (IFN)-mediated activation of antigen presenting cells, the induction of IDO and the kynurenine system exerts a counter-regulating effect, maintaining the homeostasis. Inhibition of T cell functions, activation of the regulatory T cells, and the inhibition of Natural Killer cells are among the important factors in the immunosuppressive effects of IDO and kynurenines. There is a close connection between cytokines (IFN-α, IFN-γ, TNF-α, TGF-ß, IL-4 and IL-23) and the kynurenine system, and an imbalance in the TH1/TH2 cytokine profile may possibly lead to neurologic or psychiatric disorders. As the tryptophan metabolic pathway is activated by pro-inflammatory stimuli, the anti-inflammatory effect of kynurenic acid provides a further feedback mechanism in modulating the immune responses.
Assuntos
Tolerância Imunológica/imunologia , Imunomodulação/fisiologia , Cinurenina/fisiologia , Cinurenina/uso terapêutico , Animais , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/fisiologia , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/prevenção & controle , Cinurenina/metabolismo , Vias Neurais/enzimologia , Vias Neurais/imunologia , Vias Neurais/patologia , Linfócitos T Reguladores/enzimologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/patologiaRESUMO
Kynurenic acid (KynA), a broad spectrum antagonist of excitatory amino acid receptors, may serve as a protective agent in neurological disorders. The potential anti-inflammatory effect of KynA in human leukocytes has not been characterized. The aim of this study was to compare the effects of KynA with those of a new analogue, 2-(2-N,N-dimethylaminoethylamine-1-carbonyl)-1H-quinolin-4-one hydrochloride on tumour necrosis factor-α (TNF-α) production and high mobility group box protein 1 (HMGB1) secretion. The effects of KynA on granulocyte activation were investigated via the secretion of human neutrophil peptide 1-3 (HNP1-3). Peripheral blood mononuclear cells and granulocytes or CD14 positive monocytes were applied as effector cells, or whole blood cultures were used. TNF-α, HMGB1 and HNP1-3 concentrations were determined by ELISA, TNF-α and HNP1-3 mRNA expressions were quantified by reverse transcription PCR. KynA attenuated the TNF-α production of human mononuclear cells activated by heat-inactivated Staphylococcus aureus, inhibiting TNF-α production at the transcription level. Furthermore, KynA diminished HMGB1 secretion by U 937 monocytic cells and by peripheral blood monocytes. KynA inhibited the HNP1-3 secretion in whole blood and in granulocyte cultures. The suppressive effect of the KynA analogue was more potent than that of an equimolar concentration KynA in TNF-α, HMGB1 and HNP1-3 inhibition. These results suggest that the new KynA analogue has a more potent immunoregulatory effect than KynA on human mononuclear cells, monocytes and granulocytes and indicate the potential benefits of further exploration of its uses in human inflammatory disease.
Assuntos
Proteína HMGB1/antagonistas & inibidores , Ácido Cinurênico/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Quinolonas/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , alfa-Defensinas/antagonistas & inibidores , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Proteína HMGB1/biossíntese , Humanos , Ácido Cinurênico/análogos & derivados , Leucócitos Mononucleares/imunologia , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Neutrófilos/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Necrose Tumoral alfa/biossíntese , alfa-Defensinas/metabolismoRESUMO
Defensins are an important family of natural antimicrobial peptides. Chlamydophila pneumoniae, a common cause of acute respiratory infection, has a tendency to cause persistent inflammatory diseases such as atherosclerosis, which may lead to cardiovascular disease or stroke. As endothelial cells are related to the physiopathology of stroke, the effects of in vitro C. pneumoniae infection on the expression of human ß-defensin 2 (HBD-2) in brain capillary endothelial cells (BB19) was investigated. A time-dependent increase in HBD-2 mRNA was observed by means of real-time reverse transcription PCR (RT-PCR) in BB19 cells following C. pneumoniae infection, with a maximum increase at 24 h. A gradual induction of HBD-2 protein in the C. pneumoniae-infected endothelial cells was detected by immunoblotting. Immunofluorescence revealed the staining of HBD-2 in the cytoplasm of endothelial cells following C. pneumoniae infection. The secretion of HBD-2 (confirmed by ELISA) was significantly elevated 24 h after C. pneumoniae infection. These novel results indicate that HBD-2 is expressed and produced in the human brain capillary endothelial cells upon infection with C. pneumoniae, and provide evidence that HBD-2 plays a role in the early immune responses to C. pneumoniae and probably in the immunopathogenesis of atherosclerosis.
Assuntos
Infecções por Chlamydophila/imunologia , Chlamydophila pneumoniae/imunologia , Células Endoteliais/metabolismo , Ativação Transcricional , beta-Defensinas/metabolismo , Encéfalo/irrigação sanguínea , Linhagem Celular , Infecções por Chlamydophila/complicações , Chlamydophila pneumoniae/patogenicidade , Citoplasma/metabolismo , Células Endoteliais/imunologia , Células Endoteliais/microbiologia , Células Endoteliais/patologia , Imunofluorescência , Regulação da Expressão Gênica/imunologia , Humanos , Imunidade Inata/genética , Ativação Transcricional/imunologia , beta-Defensinas/genética , beta-Defensinas/imunologiaRESUMO
BACKGROUND: The transcription factor p63 plays a pivotal role in the development and maintenance of epithelial tissues, including the ocular surface. In an effort to gain insight into the pathogenesis of keratitis caused by HSV-1, we determined the expression patterns of the p63 and Bax proteins in the Staatens Seruminstitute Rabbit Cornea cell line (SIRC). METHODS: SIRC cells were infected with HSV-1 at various multiplicities and maintained for different periods of time. Virus replication was measured by indirect immunofluorescence assay and Western blot analysis. Cell viability was determined by MTT assay. The apoptotic response of the infected cells was quantified by ELISA detecting the enrichment of nucleosomes in the cytoplasm. Western blot analysis was used to determine the levels of p63 and Bax proteins. RESULTS: Indirect immunofluorescence assays and Western blot analyses demonstrated the presence of HSV-1 glycoprotein D (gD) in the infected SIRC cell line, and the pattern of gD expression was consistent with efficient viral replication. The results of MTT and ELISA assays showed that HSV-1 elicited a strong cytopathic effect, and apoptosis played an important role in the demise of the infected cells. Mock-infected SIRC cells displayed the constitutive expression of DeltaNp63alpha. The expressions of the Bax-beta and TAp63gamma isoforms were considerably increased, whereas the level of DeltaNp63alpha was decreased in the HSV-1-infected SIRC cells. Experiments involving the use of acyclovir showed that viral DNA replication was necessary for the accumulation of TAp63gamma. CONCLUSION: These data suggest that a direct, virus-mediated cytopathic effect may play an important role in the pathogenic mechanism of herpetic keratitis. By disturbing the delicate balance between the pro-survival DeltaN and the pro-apoptotic TA isoforms, HSV-1 may cause profound alterations in the viability of the ocular cells and in the tissue homeostasis of the ocular surface.
Assuntos
Herpesvirus Humano 1/patogenicidade , Ceratite Herpética/etiologia , Proteínas Supressoras de Tumor/metabolismo , Animais , Apoptose , Linhagem Celular , Sobrevivência Celular , Efeito Citopatogênico Viral , Replicação do DNA , Herpesvirus Humano 1/fisiologia , Ceratite Herpética/metabolismo , Ceratite Herpética/patologia , Ceratite Herpética/virologia , Coelhos , Proteínas do Envelope Viral/metabolismo , Replicação Viral , Proteína X Associada a bcl-2/metabolismoRESUMO
The pathogenesis of multiple sclerosis (MS), a devastating neuroinflammatory disorder of the central nervous system, has been presumed to involve the possible importance of the receptor for advanced glycation end products (RAGE). The aim of this study was to investigate the relevance of the genetic polymorphisms of RAGE in MS patients. A total of 168 patients with MS were enrolled; 136 healthy blood donors served as controls. The -374 T/ A, -479 T/C, and the G82S polymorphisms of RAGE were determined by restriction fragment length polymorphism (RFLP). There was a significant difference in RAGE -374 T/A genotype distribution between the controls and the MS patients. The AA homozygote variants were detected in 8% of the patients with MS, as compared with 19% of healthy controls (OR=2.75; 95% CI=1.319-5.733, p = 0.007). No differences were observed between the MS patients and the controls, concerning the frequencies of the -479 T/C and G82S genotypes of the RAGE. Our results revealed an association between the -374 T/A polymorphism of the RAGE promoter and MS. The genetic variant -374 AA (which has previously been shown to exert significant effects on transcriptional activity) can be considered a preventive factor as regards the occurrence of MS. Our findings support the view that RAGE plays a role in the development of MS.
Assuntos
Predisposição Genética para Doença/genética , Esclerose Múltipla/genética , Polimorfismo Genético/genética , Receptor para Produtos Finais de Glicação Avançada/genética , Adulto , Sistema Nervoso Central/metabolismo , Sistema Nervoso Central/fisiopatologia , Análise Mutacional de DNA , Feminino , Frequência do Gene/genética , Marcadores Genéticos/genética , Testes Genéticos , Variação Genética/genética , Genótipo , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/metabolismo , Esclerose Múltipla/fisiopatologia , Fatores de Crescimento Neural/genética , Ligação Proteica/genética , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/genéticaRESUMO
BACKGROUND: The kinesin light-chain 1 genetic variants G56836C, A185C, and C406T were earlier found to amplify the development of leukoaraiosis in hypertensive smokers. These 3 variants were presumed to affect the function of the mitochondria, thereby giving rise to sensitivity to a chronic ischemic state. We have now extended our investigations to examine how the above genetic variants affect the occurrence of ischemic stroke. METHODS: Genetic and clinical data on 650 ischemic stroke and 340 neuroimaging alteration-free subjects were analyzed. Univariate and logistic regression approaches were used. RESULTS: None of the above genetic variants proved to be risk factors of ischemic stroke, either alone or in combination with other clinical factors. CONCLUSION: The examined 3 genetic variants seem to influence the responses of the glial cells to a slight chronic hypoxia state, rather than the mechanisms resulting in cerebral infarcts themselves.
Assuntos
Variação Genética/genética , Hipóxia-Isquemia Encefálica/genética , Cinesinas/genética , Acidente Vascular Cerebral/genética , Idoso , Idoso de 80 Anos ou mais , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Análise Mutacional de DNA , Feminino , Marcadores Genéticos/genética , Predisposição Genética para Doença/genética , Testes Genéticos , Humanos , Hipóxia-Isquemia Encefálica/epidemiologia , Hipóxia-Isquemia Encefálica/fisiopatologia , Masculino , Pessoa de Meia-Idade , Neuroglia/metabolismo , Consumo de Oxigênio/fisiologia , Polimorfismo Genético/genética , Fatores de Risco , Acidente Vascular Cerebral/epidemiologia , Acidente Vascular Cerebral/fisiopatologiaRESUMO
Multiple sclerosis (MS), which results in damage of the white matter at multiple foci, poses a far-reaching public health problem in view of the burden it imposes on the affected young and middle-aged. Some previous data suggested that roles could be played in the demyelinization of the white matter of the brain by the malfunctioning of the mitochondria and mitochondria-associated reactive oxygen species. In this context, we hypothesized that the finely tuned dynamic stability of the mitochondrial membrane potential (MMP), which is the main mirror of the functional state of the mitochondria, is essential for the intact nature of the glia cells in the brain. Setting out from this, our aim in this study was to examine how the rs10807344 and rs2270450 genetic variants of mitochondrial uncoupling protein 4 (mUCP4) can give rise to the development of MS, since mUCP4 is presumed to be of great importance in the regulation of the MMP and cellular energy metabolism. The clinical and genetic data on 120 relapsing-remitting MS patients and 250 neuroimaging alteration-free subjects were analyzed. The rs10807344 CC genotype proved to exert a protective effect against the occurrence of MS (neuroimaging alteration-free controls, 58%; MS group, 33%; P < 0.0000089; OR, 0.32; 95% CI: 0.2-0.56, P < 0.005). The present findings indirectly raise the possibility that a shift or imbalance in the finally regulated MMP plays a role in the development of MS.
Assuntos
Variação Genética , Proteínas de Membrana Transportadoras/genética , Esclerose Múltipla/genética , Adulto , Idoso , Feminino , Genótipo , Humanos , Masculino , Potencial da Membrana Mitocondrial/fisiologia , Proteínas de Membrana Transportadoras/metabolismo , Pessoa de Meia-Idade , Mitocôndrias/genética , Mitocôndrias/metabolismo , Proteínas de Desacoplamento MitocondrialRESUMO
p63 plays a pivotal role in the development and maintenance of stratified epithelial tissues. In an effort to gain insight into the pathogenic mechanisms of skin infections caused by HSV-1 and HSV-2, we determined the patterns of p63 expression in primary keratinocytes and in the HaCaT cell line. The levels of DeltaNp63alpha and a 50kDa p73 isoform were decreased, Bax-alpha remained unaffected, while the expressions of the Bax-beta, TAp63gamma and a 44.5kDa p73 isoform were highly increased in both HSV-1-infected HaCaT cells and primary keratinocytes. In contrast, in response to HSV-2 infection the levels of DeltaNp63alpha, a 50kDa p73 isoform and a 44.5kDa p73 protein were decreased, Bax-alpha and TAp63gamma remained unaffected, while the expression of Bax-beta was slightly increased. The knockdown of TAp63 expression enhanced the viability of HSV-1-infected cells. Thus, HSV-1 and HSV-2 modulate the patterns of p63 and Bax expression in a serotype-specific manner. The dysregulated pattern of p63 expression observed in HSV-infected keratinocytes may comprise part of a mechanism by which these viruses perturb the functions of keratinocytes and lead to their demise.