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1.
Biomed Khim ; 66(2): 162-166, 2020 Feb.
Artigo em Russo | MEDLINE | ID: mdl-32420898

RESUMO

Endometriosis is a common estrogen-dependent chronic disease in women of reproductive age; it is associated with dysregulation of the immune response, local inflammation, and increased formation of autoantibodies. The aim of the study was to investigate the profile of autoantibodies in women with endometriosis and to evaluate their diagnostic value using new modifications of enzyme immunoassay. In women with endometriosis of stage III-IV (n=39), a wide spectrum of autoantibodies was detected, mainly of class G, including antibodies to endometrial antigens (tropomyosin 3, tropomodulin 3), the enzyme α-enolase, steroid (estradiol, progesterone) and gonadotropic hormones. At the same time, the frequency of detection of IgG antibodies to tropomyosin 3, α-enolase, estradiol and human chorionic gonadotropin and their levels in patients with endometriosis were higher than in healthy women (n=26) (p<0.05). IgG-antibodies to tropomyosin 3, α-enolase and estradiol were characterized by higher diagnostic value for endometriosis. The diagnostic value was significantly increased when these antibodies were combined: the AUC reached 0.875 [0.772-0.978] (p<0.0001), the sensitivity and specificity were 83.3% each. Thus, autoantibodies to tropomyosin 3, α-enolase, and estradiol are promising for inclusion in the panel of biomarkers for non-invasive diagnosis of endometriosis.


Assuntos
Autoanticorpos/sangue , Endometriose/diagnóstico , Biomarcadores/sangue , Estradiol/imunologia , Feminino , Humanos , Fosfopiruvato Hidratase/imunologia , Tropomiosina/imunologia
2.
Immunol Lett ; 218: 22-29, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31866401

RESUMO

OBJECTIVE: Pulmonary arterial hypertension (PAH) is an intractable complication in connective tissue diseases, but the pathological mechanisms responsible for progression remain obscure. This study aims to test whether patient IgG possesses biological activity promoting the migration of pulmonary artery smooth muscle cells (PASMCs). METHODS: Cell migration was estimated by lamellipodia formation and by utilizing a Boyden chamber method. The specificity of autoantibodies was established by western blotting, ELISA, and immunocytochemistry. The target antigen was investigated by mass spectrometry. RESULTS: IgG obtained from a patient with systemic lupus erythematosus (SLE) accompanied by PAH was found to promote lamellipodia formation and migration of PASMCs. The IgG bound to a ∼50 kDa protein expressed on the cell membrane, and in the cytoplasm and nucleus. This molecule was identified as enolase 1. Removal of enolase 1-binding antibodies from the IgG fraction, or treatment of the cells with an enolase inhibitor, significantly suppressed the migration of PASMCs. CONCLUSION: Patients with SLE may possess autoantibodies to enolase 1 which stimulate the migration of PASMCs and are likely to play a role in the progression of PAH.


Assuntos
Autoanticorpos/imunologia , Biomarcadores Tumorais/imunologia , Proteínas de Ligação a DNA/imunologia , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/imunologia , Miócitos de Músculo Liso/metabolismo , Fosfopiruvato Hidratase/imunologia , Hipertensão Arterial Pulmonar/complicações , Hipertensão Arterial Pulmonar/metabolismo , Proteínas Supressoras de Tumor/imunologia , Sequência de Aminoácidos , Autoanticorpos/sangue , Autoantígenos/imunologia , Biomarcadores Tumorais/química , Biomarcadores Tumorais/metabolismo , Movimento Celular , Proliferação de Células , Células Cultivadas , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Ligantes , Miócitos de Músculo Liso/imunologia , Fosfopiruvato Hidratase/química , Fosfopiruvato Hidratase/metabolismo , Ligação Proteica , Hipertensão Arterial Pulmonar/diagnóstico , Artéria Pulmonar/citologia , Artéria Pulmonar/imunologia , Artéria Pulmonar/metabolismo , Transdução de Sinais , Proteínas Supressoras de Tumor/química , Proteínas Supressoras de Tumor/metabolismo
3.
Mol Cell Proteomics ; 19(1): 155-166, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-29089373

RESUMO

Plasmodium falciparum malaria continues to evade control efforts, utilizing highly specialized sexual-stages to transmit infection between the human host and mosquito vector. In a vaccination model, antibodies directed to sexual-stage antigens, when ingested in the mosquito blood meal, can inhibit parasite growth in the midgut and consequently arrest transmission. Despite multiple datasets for the Plasmodium sexual-stage transcriptome and proteome, there have been no rational screens to identify candidate antigens for transmission-blocking vaccine (TBV) development. This study characterizes 12 proteins from across the P. falciparum sexual-stages as possible TBV targets. Recombinant proteins are heterologously expressed as full-length ectodomains in a mammalian HEK293 cell system. The proteins recapitulate native parasite epitopes as assessed by indirect fluorescence assay and a proportion exhibits immunoreactivity when tested against sera from individuals living in malaria-endemic Burkina Faso and Mali. Purified IgG generated to the mosquito-stage parasite antigen enolase demonstrates moderate inhibition of parasite development in the mosquito midgut by the ex vivo standard membrane feeding assay. The findings support the use of rational screens and comparative functional assessments in identifying proteins of the P. falciparum transmission pathway and establishing a robust pre-clinical TBV pipeline.


Assuntos
Anticorpos Bloqueadores/imunologia , Malária Falciparum/imunologia , Malária Falciparum/transmissão , Plasmodium falciparum/imunologia , Proteínas de Protozoários/imunologia , Proteínas Recombinantes/imunologia , Adulto , Animais , Anopheles/parasitologia , Epitopos/imunologia , Feminino , Células HEK293 , Humanos , Imunoglobulina G/imunologia , Vacinas Antimaláricas/imunologia , Malária Falciparum/epidemiologia , Malária Falciparum/virologia , Masculino , Mali/epidemiologia , Camundongos , Camundongos Endogâmicos BALB C , Mosquitos Vetores/parasitologia , Fosfopiruvato Hidratase/imunologia , Proteoma , Proteômica/métodos , Vacinação
4.
Clin Immunol ; 210: 108317, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31770612

RESUMO

Autoantibodies (AAbs) against retinal antigens can be found in patients with cancer and unexplained vision loss unrelated to the cancer metastasis. Cancer-associated retinopathy (CAR) is a rare paraneoplastic visual syndrome mediated by AAbs. Our goal was to determine whether CAR patients with different malignancies have a specific AAb or repertoire of AAbs that could serve as biomarkers for retinal disease. We found AAbs against 12 confirmed retinal antigens, with α-enolase being the most frequently recognized. The significant finding of the study was a high incidence of anti-aldolase AAbs in colon-CAR, anti-CAII in prostate-CAR, and anti-arrestin in skin melanoma patients thus these AAbs could serve as biomarkers in the context of clinical presentation and could support the diagnosis of CAR. However, a lack of AAb restriction to any one antigenic protein or to one retinal cellular location makes screening for a CAR biomarker challenging.


Assuntos
Autoanticorpos/sangue , Biomarcadores Tumorais/sangue , Neoplasias do Colo/imunologia , Síndromes Paraneoplásicas Oculares/imunologia , Neoplasias da Próstata/imunologia , Retina/patologia , Idoso , Arrestina/imunologia , Autoantígenos/imunologia , Neoplasias do Colo/diagnóstico , Neoplasias do Colo/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Síndromes Paraneoplásicas Oculares/diagnóstico , Síndromes Paraneoplásicas Oculares/epidemiologia , Fosfopiruvato Hidratase/imunologia , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/epidemiologia , Retina/imunologia , Estados Unidos/epidemiologia
5.
Ann Clin Lab Sci ; 49(4): 503-506, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31471340

RESUMO

GOALS: Rheumatoid arthritis (RA) is a chronic inflammatory joint disease. The α enolase is a nuclear glycolytic enzyme. Antibodies to citrullinated-enolase peptide1(anti-CEP-1) are found in approximately 40% of patients with RA, but the diagnostic value of anti-CEP-1 for RA is not clear. METHODS: We enrolled 282 patients with RA according to the 2010 ACR Classification criteria, referred to the department of Rheumatology in Peking University Third Hospital, 120 sex- and age-matched healthy donors (HD), and 30 patients with osteoarthritis (OA). Anti-CEP-1 IgG antibodies were assessed with a commercially available ELISA kit (Euroimmun, Germany) according to the manufacturers' instructions. Anti-CCP antibodies were assessed with ECLIA (Roche, Germany). Data was processed with SPSS 19. The scatter diagram was drawn in GraphPad Prism. RESULTS: The specificity and sensitivity was 83.3% and 65.2%, respectively. The positive predictive value was 88% and the negative predictive value was 56%. The AUC of anti-CEP1 for diagnosis of RA is 0.80, while that of Anti-CCP is 0.919; the value of anti-CCP combined with anti-CEP1 is 0.914. Ten anti-CEP1 positive results are found in 48 patients of RA with anti-CCP negative result. CONCLUSION: The anti-CEP-1 is suitable for diagnosing of RA, but not superior to anti-CCP.


Assuntos
Anticorpos Anti-Proteína Citrulinada/imunologia , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/imunologia , Peptídeos/imunologia , Fosfopiruvato Hidratase/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Adulto Jovem
6.
Analyst ; 144(16): 4813-4819, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31281909

RESUMO

In the clinical diagnosis of tumor, the immunological detection of single tumor markers may lead to errors and missed inspection. Therefore, it is necessary to establish an accurate and effective method for the simultaneous detection of multiple tumor markers. Thus, we developed a time-resolved chemiluminescence immunoassay (TRCLIA) to simultaneously detect carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in human serum. Horseradish peroxidase (HRP) and alkaline phosphatase (ALP) were used as the detection probes to label the monoclonal antibodies of CEA and NSE by strain-promoted azide-alkyne cycloaddition (SPAAC), respectively. Based on a sandwich immunoassay, the targets in the samples were captured by antibodies immobilized on the surface of carboxylate-modified polystyrene microspheres (CPSMS) and sandwiched by other antibodies labeled with HRP and ALP. Since HRP and ALP had different dynamic characteristics, the CEA and NSE signals were recorded at 0.5 s and 20 min, respectively, and cross-interference could be avoided effectively. The whole signal detection processes could be completed in 20 min. The linear ranges of CEA and NSE were 0.1-64 ng mL-1 and 0.05-64 ng mL-1 and the limits of detection were 0.085 ng mL-1 and 0.044 ng mL-1 (S/N = 2), respectively. Also, 45 human serum samples obtained from patients having lung disease were tested by TRCLIA and commercial chemiluminescence enzyme-linked immunoassay (CLEIA) kits with good correlation. The correlation coefficients of CEA and NSE were 0.985 and 0.970, respectively. The results demonstrated a novel, effective, reliable and convenient TRCLIA method for the clinical diagnosis of CEA and NSE. The TRCLIA method has the potential to be an effective clinical tool for the early screening of lung cancer and can be applied in clinical diagnosis.


Assuntos
Antígeno Carcinoembrionário/sangue , Técnicas Imunoenzimáticas/métodos , Fosfopiruvato Hidratase/sangue , Fosfatase Alcalina/química , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais/imunologia , Armoracia/enzimologia , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/imunologia , Antígeno Carcinoembrionário/imunologia , Peroxidase do Rábano Silvestre/química , Humanos , Limite de Detecção , Luminescência , Substâncias Luminescentes/química , Medições Luminescentes/métodos , Luminol/química , Pneumopatias/sangue , Fosfopiruvato Hidratase/imunologia
7.
Graefes Arch Clin Exp Ophthalmol ; 257(8): 1751-1758, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31065846

RESUMO

PURPOSE: To compare the clinical characteristics of Vogt-Koyanagi-Harada (VKH) disease patients with and without anti-retinal antibodies (ARAs) that are frequently detected in autoimmune retinopathy. METHODS: Using immunoblot analyses, serum autoantibodies for recoverin, carbonic anhydrase II, and α-enolase were examined in 20 treatment-naïve patients with VKH disease. Clinical factors before and after systemic corticosteroid therapy, including best-corrected visual acuity (BCVA) and macular outer retinal morphology, were statistically compared between patients with VKH disease with and without ARAs. RESULTS: Serum ARAs were detected in 50.0% of patients with VKH disease. There were no significant differences in clinical factors between the two groups, including final BCVA, frequency of uveitis recurrence, and recovery of the macular ellipsoid zone after systemic corticosteroid therapy. CONCLUSIONS: Our results suggest that the detected ARAs did not influence visual outcomes, the chronicity of uveitis, or outer retinal morphology in patients with VKH disease.


Assuntos
Autoanticorpos/imunologia , Retina/imunologia , Síndrome Uveomeningoencefálica/imunologia , Acuidade Visual , Adolescente , Adulto , Autoanticorpos/sangue , Anidrase Carbônica II/sangue , Anidrase Carbônica II/imunologia , Criança , Feminino , Glucocorticoides/uso terapêutico , Humanos , Immunoblotting , Masculino , Pessoa de Meia-Idade , Fosfopiruvato Hidratase/sangue , Fosfopiruvato Hidratase/imunologia , Prognóstico , Recoverina/sangue , Recoverina/imunologia , Retina/diagnóstico por imagem , Estudos Retrospectivos , Tomografia de Coerência Óptica , Síndrome Uveomeningoencefálica/tratamento farmacológico , Adulto Jovem
8.
Graefes Arch Clin Exp Ophthalmol ; 257(8): 1759-1764, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31119427

RESUMO

PURPOSE: To explore the presence of serum anti-retinal antibodies (ARAs) in the Chinese patients with presumed autoimmune retinopathy (AIR). METHODS: Twenty-three Chinese patients with presumed AIR, disease controls including 40 RP patients, 22 bilateral uveitis patients, 18 acute zonal outer occult retinopathy (AZOOR) patients, and 30 healthy donors were included. Serum samples of all the subjects were obtained and analyzed for the presence of four ARAs including recoverin, α-enolase, carbonic anhydraseII (CAII), and collapsin response-mediated protein (CRMP)-5 by Western bolt assay. RESULTS: ARAs were present in the serum of either presumed AIR patients, disease control, or healthy donors. One or more ARAs were present in the 78.2% of presumed AIR while they were indicated in the 35.0% of RP patients (p < 0.01) and 33.3% of healthy donors (p < 0.01). The prevalence of ARAs in the bilateral uveitis and AZOOR was 63.3% and 100% respectively. Positive rate of α-enolase antibody present in the presumed AIR, disease control, and healthy donors was 73.9%, 47.5%, and 33.3% respectively. Positive rate of CAII antibody present above groups was 52.1%, 50%, and 33.3% respectively. Recoverin antibody seemed to be specifically present in the serum of patients with cancer-associated retinopathy. CONCLUSION: Presence of serum ARAs including recoverin, α-enolase, CAII, or CRMP-5 in the Chinese patients with presumed AIR occurred significantly more often than RP patients and healthy donors. Seropositivity of ARAs had diagnostic value for the presumed AIR but mere presence was not sufficient for the diagnosis due to identification of them in the healthy controls and other retinal diseases.


Assuntos
Autoanticorpos/sangue , Doenças Autoimunes/imunologia , Retina/imunologia , Doenças Retinianas/imunologia , Adulto , Idoso , Doenças Autoimunes/sangue , Doenças Autoimunes/epidemiologia , Western Blotting , Anidrase Carbônica II/sangue , Anidrase Carbônica II/imunologia , China/epidemiologia , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Proteínas do Tecido Nervoso/sangue , Proteínas do Tecido Nervoso/imunologia , Fosfopiruvato Hidratase/sangue , Fosfopiruvato Hidratase/imunologia , Prevalência , Recoverina/sangue , Recoverina/imunologia , Doenças Retinianas/sangue , Doenças Retinianas/epidemiologia , Estudos Retrospectivos
9.
Exp Parasitol ; 200: 92-98, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30991039

RESUMO

Adult Brugia malayi proteins with high potential as epidemiological markers, diagnostic and therapeutic targets, and/or vaccine candidates were revealed by using microfilaremic human sera and an immunoproteomic approach. They were HSP70, cytoplasmic intermediate filament protein, independent phosphoglycerate mutase, and enolase. Brugia malayi microfilaria-specific proteins that formed circulating immune complexes (ICs) were investigated. The IC-forming proteins were orthologues of hypothetical protein Bm1_12480, Pao retrotransposon peptidase family protein, uncoordinated protein 44, NAD-binding domain containing protein of the UDP-glucose/GDP-mannose dehydrogenase family which contained ankyrin repeat region, ZU5 domain with C-terminal death domain, C2 domain containing protein, and FLJ90013 protein of the eukaryotic membrane protein family. Antibodies to these proteins were not free in the microfilaremic sera, raising the possible role of the IC-forming proteins in an immune evasion mechanism of the circulating microfilariae to avoid antibody-mediated-host immunity. Moreover, detection of these ICs should be able to replace the inconvenient night blood sampling for microfilaria in an evaluation of efficacy of anti-microfilarial agents.


Assuntos
Complexo Antígeno-Anticorpo/imunologia , Antígenos de Helmintos/imunologia , Brugia Malayi/imunologia , Filariose/imunologia , Proteínas de Helminto/imunologia , Soros Imunes/imunologia , Animais , Biologia Computacional , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Filariose/sangue , Proteínas de Choque Térmico HSP70/imunologia , Humanos , Immunoblotting , Proteínas de Filamentos Intermediários/imunologia , Microfilárias/imunologia , Fosfoglicerato Mutase/imunologia , Fosfopiruvato Hidratase/imunologia , Proteômica/métodos
10.
EBioMedicine ; 41: 610-622, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30827932

RESUMO

BACKGROUND: We recently demonstrated the increased abundance of anti-trophoblast antibodies (ATAB) in sera of patients with unexplained recurrent miscarriages (uRM). Further, the ATAB-positive sera bound to JEG-3 human choriocarcinoma cells in vitro, resulting in decreased productions of ß-human chorionic gonadotropin (ß-hCG) and progesterone in these cells. However, the specific antigenic epitopes of ATAB have remained unknown. Therefore, it was the aim of this study to determine specific targets of ATAB in uRM patients. METHODS: Potential targets of ATAB were analyzed by 2-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry, and thereby identifying α-Enolase (ENO1). ATAB targeting of ENO1 was further confirmed in a competitive binding assay. Levels of anti-ENO1 antibodies as well as ß-hCG and progesterone were quantified with enzyme-linked immunosorbent assay (ELISA). Additionally, expression of ENO1 was analyzed in first trimester placentas by immunohistochemistry and immunofluorescence analysis. FINDINGS: We here identified ENO1 as a prominent target of ATAB. Serum levels of anti-ENO1 antibodies were increased in ATAB-positive compared to ATAB-negative patients. Further, increased expression of ENO1 and its co-expression with ß-arrestin was found in the extra villous trophoblasts of uRM patients in first trimester placentas. In vitro, anti-ENO1 antibodies decreased the secretion of ß-hCG and progesterone in JEG-3 and primary human villous trophoblast cells. INTERPRETATION: Serum anti-ENO1 antibodies might be an autoimmune biomarker for uRM. Targeting the formation of anti-ENO1 antibodies or inhibition of ENO1 expression could potentially represent therapeutic strategies for these patients. FUND: All authors declare no conflict of interest. Yao Ye was supported by the China Scholarship Council. Hellen Ishikawa-Ankerhold and Christian Schulz were supported by the SFB914, projects Z01 and A10. None of the rest authors has any conflict of interest to declare.


Assuntos
Aborto Habitual/diagnóstico , Autoanticorpos/sangue , Doenças Autoimunes/diagnóstico , Biomarcadores/sangue , Fosfopiruvato Hidratase/imunologia , Aborto Habitual/patologia , Linhagem Celular Tumoral , Gonadotropina Coriônica/análise , Gonadotropina Coriônica/metabolismo , Citocinas , Eletroforese em Gel Bidimensional , Feminino , Humanos , Espectrometria de Massas , Microscopia Confocal , Placenta/metabolismo , Placenta/patologia , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Gravidez , Progesterona/análise , Progesterona/metabolismo , Trofoblastos/citologia , Trofoblastos/imunologia , Trofoblastos/patologia
11.
Lupus ; 28(3): 365-370, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30813870

RESUMO

OBJECTIVE: Anti-α-enolase antibody (Ab) combined with ß2 microglobulin (ß2-MG) were investigated to predict the incidence of nephritis in systemic lupus erythematosus (SLE) patients. METHODS: Levels of serum anti-α-enolase Ab and urinary ß2-MG were detected in 115 SLE patients, 29 SLE patients with nephritis and 70 healthy controls by ELISA and immunoturbidimetry, respectively. Furthermore, the correlation between anti-α-enolase Ab combined with ß2-MG and the incidence of nephritis in SLE patients was evaluated by correlation analysis. RESULTS: The optical density value of serum anti-α-enolase Ab in SLE patients with nephritis (0.84) was greatly increased compared with SLE patients (0.76) or healthy controls (0.54). Moreover, the levels of urinary ß2-MG in SLE patients with nephritis (6.75 mg/L) were increased compared with SLE patients (3.45 mg/L) or healthy controls (1.48 mg/L). There was a positive correlation between the level of anti-α-enolase Ab and ß2-MG ( r = 0.754). Furthermore, anti-α-enolase Ab combined with ß2-MG for evaluating the incidence of nephritis in SLE patients had the best assessment of the effectiveness (area under the receiver operating characteristic curve (AUC): 92.7%) compared with only anti-α-enolase Ab (AUC: 80.9%) or ß2-MG (AUC: 84.5%). CONCLUSION: These data suggested that anti-α-enolase Ab may be a potential indicator for the prediction of nephritis in SLE patients.


Assuntos
Nefrite Lúpica/sangue , Nefrite Lúpica/urina , Fosfopiruvato Hidratase/sangue , Microglobulina beta-2/urina , Adulto , Área Sob a Curva , Biomarcadores/sangue , Biomarcadores/urina , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Nefrite Lúpica/diagnóstico , Masculino , Pessoa de Meia-Idade , Fosfopiruvato Hidratase/imunologia , Valor Preditivo dos Testes , Microglobulina beta-2/imunologia
12.
Analyst ; 144(6): 2186-2194, 2019 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-30785140

RESUMO

This work was aimed at designing a novel and ultrasensitive electrochemical immunoassay strategy to detect neuron-specific enolase (NSE) with a triple signal amplification strategy. A greatly enhanced sensitive detection of NSE was achieved by using porous three-dimensional graphene-starch architecture (3D-GNS) modified on the immunosensor's surface to construct a unique 3D immunoelectrode, which would greatly accelerate electron transfer and capture more protein molecules. 3D-GNS was prepared with starch as a crosslinking agent and stabilizer, which is biocompatible and environmentally friendly. Aggregation-free gold nanoparticle (AuNP) incorporated ordered mesoporous carbon-silica (OMCSi-Au) with good catalytic activity was synthesized as the tracing tag for labeling signal antibody (Ab2). After a sandwich-type immunoreaction, the OMCSi-Au labeled Ab2 was trapped on the surface of the immunosensor, and the high concentration of AuNPs with high dispersion greatly catalyzed the deposition of silver nanoparticles. The deposited silver nanoparticles (AgNPs) could be tested directly with anodic stripping voltammetric analysis (ASV) in potassium chloride solution to monitor the immunoreactions, which greatly enhanced the sensitivity of protein markers with a detection limit of 0.008 pg mL-1, and a linear range of 0.02 pg mL-1 to 35 ng mL-1 for neuron-specific enolase antigen. This proposed immunosensor displayed acceptable accuracy, good stability, and high sensitivity. Good results were also obtained in agreement with the enzyme-linked immunosorbent assay method, which provides greatly promising potential in clinical applications.


Assuntos
Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Grafite/química , Neoplasias Pulmonares/sangue , Fosfopiruvato Hidratase/sangue , Prata/química , Amido/química , Anticorpos Monoclonais/imunologia , Ouro/química , Voluntários Saudáveis , Humanos , Imunoensaio , Limite de Detecção , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/imunologia , Nanopartículas Metálicas/química , Fosfopiruvato Hidratase/imunologia , Porosidade , Dióxido de Silício/química
13.
Rinsho Shinkeigaku ; 59(2): 102-104, 2019 Feb 23.
Artigo em Japonês | MEDLINE | ID: mdl-30700687

RESUMO

We report a 77-year-old woman suffering from dementia with Lewy bodies (DLB) who presented with Hashimoto encephalopathy. The patient began to mistakenly believe that another person was sleeping in her bed from approximately 70 years of age. She began to show symptoms of parkinsonism after 75 years of age. One night, the patient began to exhibit loitering behavior, and made incomprehensible comments while also exhibiting other abnormal behaviors. Clinical examination revealed rigidity and tremor of the limbs, as well as hallucination, abnormal speech and behavior. We first considered DLB. However, serum anti-thyroglobulin levels turned out to be elevated, indicating Hashimoto encephalopathy as well, and treated the patient with steroid pulse therapy. Her mental symptoms subsequently improved, but rigidity and tremor remained. 123I-ioflupane SPECT demonstrated decreased accumulation in the bilateral caudal basal ganglia. Anti NH2-terminal of α-enolase (NAE) antibody in the serum was positive. Therefore, we diagnosed the patient with the rare comorbidity of DLB and Hashimoto encephalopathy, successfully treated with immunotherapy.


Assuntos
Encefalite/tratamento farmacológico , Doença de Hashimoto/tratamento farmacológico , Imunoterapia/métodos , Doença por Corpos de Lewy/tratamento farmacológico , Metilprednisolona/administração & dosagem , Prednisolona/administração & dosagem , Idoso , Autoanticorpos/sangue , Biomarcadores/sangue , Encefalite/complicações , Encefalite/diagnóstico , Feminino , Doença de Hashimoto/complicações , Doença de Hashimoto/diagnóstico , Humanos , Radioisótopos do Iodo , Doença por Corpos de Lewy/complicações , Doença por Corpos de Lewy/diagnóstico por imagem , Nortropanos , Fosfopiruvato Hidratase/imunologia , Pulsoterapia , Compostos Radiofarmacêuticos , Tomografia Computadorizada de Emissão de Fóton Único , Resultado do Tratamento
14.
Sci Immunol ; 4(31)2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30683669

RESUMO

In the context of solid tumors, there is a positive correlation between the accumulation of cytotoxic CD8+ tumor-infiltrating lymphocytes (TILs) and favorable clinical outcomes. However, CD8+ TILs often exhibit a state of functional exhaustion, limiting their activity, and the underlying molecular basis of this dysfunction is not fully understood. Here, we show that TILs found in human and murine CD8+ melanomas are metabolically compromised with deficits in both glycolytic and oxidative metabolism. Although several studies have shown that tumors can outcompete T cells for glucose, thus limiting T cell metabolic activity, we report that a down-regulation in the activity of ENOLASE 1, a critical enzyme in the glycolytic pathway, represses glycolytic activity in CD8+ TILs. Provision of pyruvate, a downstream product of ENOLASE 1, bypasses this inactivity and promotes both glycolysis and oxidative phosphorylation, resulting in improved effector function of CD8+ TILs. We found high expression of both enolase 1 mRNA and protein in CD8+ TILs, indicating that the enzymatic activity of ENOLASE 1 is regulated posttranslationally. These studies provide a critical insight into the biochemical basis of CD8+ TIL dysfunction.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Glucose/metabolismo , Linfócitos do Interstício Tumoral/imunologia , Melanoma/imunologia , Melanoma/metabolismo , Fosfopiruvato Hidratase/metabolismo , Animais , Antineoplásicos Imunológicos/uso terapêutico , Linhagem Celular Tumoral , Transportador de Glucose Tipo 1/metabolismo , Glicólise , Receptor Celular 2 do Vírus da Hepatite A/antagonistas & inibidores , Humanos , Imunoglobulina G/uso terapêutico , Imunoterapia Adotiva , Melanoma/genética , Melanoma/terapia , Camundongos Endogâmicos C57BL , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/imunologia , Receptor de Morte Celular Programada 1/antagonistas & inibidores
15.
Clin Immunol ; 200: 10-15, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30611755

RESUMO

We evaluated the clinical performance of anti-CEP-1 in a Chinese rheumatoid arthritis (RA) cohort. A total of 264 subjects were tested, including 101 RA patients, 38 juvenile idiopathic arthritis (JIA) patients, 46 disease control (DC) and 79 healthy controls (HC). The presence of anti-CEP-1 in patients with RA, JIA, DCs and HC were 61.4%, 13.2%, 15.2% and 5.1%, respectively. Anti-CCP2 demonstrated the highest positive likelihood ratio of 10.11 in the diagnosis of RA, followed by RF (8.88) and anti-CEP-1 (5.82). Anti-CEP-1 positive RA patients displayed significantly higher DAS28 compared to anti-CEP-1 negative RA patients (p = .045). Significant associations were identified between anti-CEP-1 and joint erosions at anti-CEP-1 value of >124.78 U/ml (p = .0026) and between anti-CEP-1 and ILD at anti-CEP-1 value of >185.91 U/ml (p = .0222). Our findings indicate that anti-CEP-1 may not be able to replace anti-CCP2 for routine diagnosis for RA, but they may be helpful for subtyping of the disease.


Assuntos
Anticorpos Anti-Proteína Citrulinada/imunologia , Artrite Reumatoide/imunologia , Grupo com Ancestrais do Continente Asiático , Doenças Pulmonares Intersticiais/imunologia , Fosfopiruvato Hidratase/imunologia , Adolescente , Adulto , Idoso , Artrite Juvenil/imunologia , Autoanticorpos , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fator Reumatoide/imunologia , Adulto Jovem
16.
Clin Rheumatol ; 38(3): 827-834, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30406565

RESUMO

OBJECTIVES: The aim of the study was to evaluate the frequency of anti-mutated citrullinated vimentin antibodies (a-Sa), anti-citrullinated α-enolase peptide 1 antibodies (a-CEP-1), anti-filaggrin antibodies (AFAs), heterogeneous nuclear ribonucleoprotein compies/anti-RA33-antibodies (a-hnRNP/RA33), anti-carbamylated protein antibodies (a-CarP), and metalloproteinase (MMPs) activity in patients with early inflammatory arthritis (EIA). METHODS: Seventy-four patients with EIA: 51 diagnosed with RA (rheumatoid arthritis) and 23 with UA (undifferentiated arthritis), and 20 healthy volunteers were enrolled to the study. Inflammatory markers, rheumatoid factor (RF), and antibodies mentioned above were assessed in all patients. RESULTS: In the EIA group, we observed significantly higher concentration of a-CEP-1 (65.8 ± 111.6 RU/mL) than in controls (2.0 ± 0.0 RU/mL). In RF(+) RA patients, we observed higher concentration of a-Sa and a-CEP-1 than in other groups. A-Sa were positive in 69% of RF(+) RA, 37% of RF(-) RA, 26% of UA patients and in 10% of controls. A-CEP-1 were positive in 77% of RF(+) RA patients, in 56% of RF(-) RA patients, in 8.7% of UA patients, but they were negative in controls. In patients with RF(+) RA, positive a-CarP were present statistically significantly more often than in RF (-) RA patients. No statistically significant difference in frequency of a-hnRNP/RA33 and AFA between RF(+) RA, RF(-) RA, and UA was observed. CONCLUSIONS: Our results suggest that a-CEP-1 may help in differentiation between RF(-) RA and UA. a-CEP-1 and a-Sa may be useful while diagnosing EIA. a-CarP may be used in differentiation of RA RF(-) and UA. However, a follow-up study is needed to evaluate the prognostic value of analyzed antibodies.


Assuntos
Artrite Reumatoide/imunologia , Autoanticorpos/imunologia , Metaloproteinases da Matriz Secretadas/metabolismo , Adulto , Idoso , Anticorpos Anti-Proteína Citrulinada/imunologia , Artrite/imunologia , Artrite/metabolismo , Artrite Reumatoide/metabolismo , Biomarcadores Tumorais/imunologia , Estudos de Casos e Controles , Proteínas de Ligação a DNA/imunologia , Feminino , Ribonucleoproteínas Nucleares Heterogêneas/imunologia , Humanos , Proteínas de Filamentos Intermediários/imunologia , Masculino , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , Fosfopiruvato Hidratase/imunologia , Carbamilação de Proteínas , Fator Reumatoide/imunologia , Proteínas Supressoras de Tumor/imunologia
17.
Clin Exp Rheumatol ; 37 Suppl 118(3): 29-35, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30418121

RESUMO

OBJECTIVES: Although anti-cyclic citrullinated peptides antibodies are specific markers for rheumatoid arthritis (RA), they might be present in other diseases. Our aim was to assess the native or citrullinated antigens recognised by patients with primary Sjögren's syndrome (pSS) and to evaluate their association with clinical and serological features. METHODS: In an initial screening, we assessed the serum reactivity of 12 patients with pSS against native or in vitro citrullinated antigens of HEp-2 cells by immunoblotting. We identified a 47kDa band, which was preferentially recognised and corresponded to α-enolase. Thus, levels of IgA and IgG anti-native and citrullinated α-enolase antibodies were measured in 50 pSS patients, 20 RA patients and 20 healthy subjects (HS) by ELISA. RESULTS: We identified α-enolase as a predominant antigen recognised in pSS. These patients had higher levels of anti-citrullinated α-enolase IgG antibodies compared with RA or HS (p=0.003 and p<0.0001, respectively). Furthermore, there was an increase of IgG anti-citrullinated α-enolase vs IgG anti-non-citrullinated α-enolase antibodies in pSS patients (p=0.001), by contrast no difference was found in RA. The presence of IgA and IgG anti-non-citrullinated and anti-citrullinated α-enolase antibodies were not associated with any clinical manifestation whatsoever, including non-erosive arthritis among pSS, but an association of IgA anti-citrullinated α-enolase with anti-Ro/SSA antibodies was found. CONCLUSIONS: We characterised α-enolase as a dominant antigen in lysates of HEp- 2 cells in pSS. Nevertheless, their precise role in pSS remains to be elucidated.


Assuntos
Anticorpos Anti-Proteína Citrulinada/imunologia , Fosfopiruvato Hidratase/imunologia , Síndrome de Sjogren/imunologia , Especificidade de Anticorpos , Autoanticorpos , Humanos , Peptídeos Cíclicos , Síndrome de Sjogren/enzimologia
18.
Artigo em Inglês | MEDLINE | ID: mdl-30333963

RESUMO

Three Streptococcus agalactiae (group B streptococci, GBS) immunoreactive proteins: enolase (47.4 kDa), inosine 5'-monophosphate dehydrogenase (IMPDH) (53 kDa) and molecular chaperone GroEL (57 kDa) were subjected to investigation. Enolase protein was described in our previous paper, whereas IMPDH and GroEL were presented for the first time. The aim of our paper was to provide mapping of specific epitopes, highly reactive with umbilical cord blood serum. Bioinformatic analyses allowed to select 32 most likely epitopes for enolase, 36 peptides for IMPDH and 41 immunoreactive peptides for molecular chaperone GroEL, which were synthesized by PEPSCAN. Ten peptides: two in enolase, one in IMPDH and seven in molecular chaperone GroEL have been identified as potentially highly selective epitopes that can be used as markers in rapid immunological diagnostic tests or constitute a component of an innovative vaccine against GBS infections.


Assuntos
Chaperonina 60/imunologia , Mapeamento de Epitopos , Epitopos/imunologia , IMP Desidrogenase/imunologia , Fosfopiruvato Hidratase/imunologia , Streptococcus agalactiae/imunologia , Anticorpos Antibacterianos/sangue , Biologia Computacional , Testes Diagnósticos de Rotina/métodos , Imunoensaio/métodos , Infecções Estreptocócicas/diagnóstico
19.
Am J Ophthalmol ; 196: 181-196, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30195891

RESUMO

PURPOSE: To evaluate clinical features of Japanese patients with anti-α-enolase antibody-positive autoimmune retinopathy (anti-enolase AIR). DESIGN: Multicenter retrospective observational case series. METHODS: Forty-nine eyes of 25 Japanese anti-enolase AIR patients (16 female and 9 male; mean age at first visit, 60.8 years) were included. Fundus characteristics, perimetry, spectral-domain optical coherence tomography (SD-OCT), electroretinography (ERG), best-corrected visual acuity (BCVA), and complicating systemic tumors were assessed. Protein localization of α-enolase was examined by immunohistochemistry in an enucleated eye of 1 patient. RESULTS: Patients were classified into 3 groups: multiple drusen (48%), retinal degeneration (36%), and normal fundus (16%). Drusen varied in size from small deposits to vitelliform-like lesions. Images on SD-OCT revealed dome-shaped hyperreflectivity beneath the retinal pigment epithelium (RPE), corresponding to drusen. Perimetry showed that ring scotoma was the most frequent (39%). Rod-system and/or single-flash cone responses revealed decreased responses in 81% of the eyes. Combined rod and cone system responses demonstrated significantly lower a-wave amplitudes in the degeneration group than in the drusen group (P = .005). BCVA was improved or maintained in 80% of the eyes during follow-up. Malignant or benign tumors were detected in 30% of patients. The RPE and photoreceptor layers were immunopositive for α-enolase. CONCLUSIONS: The drusen subtype, scarcely described in the literature, is suggested to characterize Japanese patients with anti-enolase AIR. The different funduscopic features with different functional severities may have resulted from antibody-mediated damage to RPE as well as photoreceptor cells.


Assuntos
Doenças Autoimunes/patologia , Fosfopiruvato Hidratase/imunologia , Drusas Retinianas/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anidrase Carbônica II/imunologia , Eletrorretinografia , Feminino , Humanos , Imuno-Histoquímica , Japão , Masculino , Pessoa de Meia-Idade , Fosfopiruvato Hidratase/metabolismo , Recoverina/imunologia , Drusas Retinianas/imunologia , Estudos Retrospectivos , Tomografia de Coerência Óptica , Adulto Jovem
20.
PLoS One ; 13(8): e0203214, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30161253

RESUMO

OBJECTIVE: Peptidylarginine deiminase 2 (PAD2) and PAD4 are expressed in the synovium of rheumatoid arthritis (RA) patients and catalyze citrullination of arginine residues in proteins targeted by anti-citrullinated protein antibodies (ACPAs). Little is known about the relative importance of PAD2 and PAD4 in generating citrullinated self-antigens. Here we investigate the ability of PAD2 and PAD4 to generate citrullinated targets for ACPAs in four human proteins. METHODS: Synovial fluid (SF) and plasma were collected from 42 RA patients. Human fibrinogen, human alpha-enolase (ENO1), human histone H3, and human serum albumin (HSA) were citrullinated in vitro by PAD2 or PAD4. The total degree of citrullination was determined using the anti-modified citrulline approach. Antibody binding to native and citrullinated proteins was measured by ELISA. RESULTS: ACPAs within pooled SF from multiple RA patients reacted equally well with, and cross-reacted with, PAD2- and PAD4-citrullinated fibrinogen. ACPAs from most individual patient SF and plasma samples bound equally well to PAD2- and PAD4-citrullinated fibrinogen or ENO1. When histone H3 was used as target, PAD4 was generally superior in generating epitopes recognized by ACPAs. No binding to citrullinated HSA was observed. CONCLUSION: In most patients, PAD2 and PAD4 are equally efficient in generating citrullinated target sites for ACPAs in fibrinogen and ENO1. The binding of autoantibodies to histone H3 was generally higher after citrullination with PAD4 than with PAD2. Citrullinated HSA is not a target for ACPAs.


Assuntos
Anticorpos Anti-Proteína Citrulinada/metabolismo , Artrite Reumatoide/enzimologia , Artrite Reumatoide/imunologia , Desiminases de Arginina em Proteínas/metabolismo , Cálcio/metabolismo , Citrulinação , Fibrinogênio/imunologia , Histonas/imunologia , Humanos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Fosfopiruvato Hidratase/imunologia , Proteína-Arginina Desiminase do Tipo 2 , Proteína-Arginina Desiminase do Tipo 4 , Proteínas Recombinantes/metabolismo , Albumina Sérica/imunologia , Líquido Sinovial
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