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1.
Front Immunol ; 13: 890502, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35707546

RESUMO

Objective: The aim of the research is to study the association between the serum levels of autoantibodies against one important epitope (168FMILPVGAANFREAMR183, designated as P6) of α-enolase (ENO1-P6Abs) and miscarriage among euthyroid females with thyroid autoimmunity (TAI). Methods: Anti-ENO1-P6 total IgG was investigated in 432 euthyroid women, and its four subclasses were analyzed in 184 euthyroid women. The serum FT4, TSH, TgAb, and TPOAb levels were determined using an electrochemiluminescence immunoassay. The serum ENO1-P6Ab and anti-protein disulfide isomerase A3 autoantibody (PDIA3Ab) levels were determined using an enzyme-linked immunosorbent assay. Results: The serum levels of anti-ENO1-P6 total IgG, IgG2, IgG3, and IgG4 were significantly higher in euthyroid TAI females than in non-TAI controls. Additionally, anti-ENO1-P6 total IgG and its 4 subtypes were all markedly higher in euthyroid TAI females with pregnancy loss than those without miscarriage. Moreover, logistic regression analysis showed that highly expressed anti-ENO1-P6 total IgG, IgG1, IgG2, and IgG3 subtypes in the serum were all independent risk factors for euthyroid TAI-related miscarriage, and its IgG1 was also for non-TAI-related abortion. According to the trend test, the prevalence of miscarriage was increased in a titer-dependent manner with the raised levels of serum anti-ENO1-P6 total IgG and IgG1, IgG2, and IgG3 subtypes among euthyroid TAI females. The receiver operating characteristic curve analysis of anti-ENO1-P6 total IgG and IgG1, IgG2, and IgG3 subclass expressions in the serum for miscarriage prediction in euthyroid TAI females exhibited that the total areas under the curves were 0.773 ± 0.041, 0.761 ± 0.053, 0.827 ± 0.043, and 0.760 ± 0.050, respectively (all P <0.0001). Their corresponding optimal cut-off OD450 values were 0.68 (total IgG), 0.26 (IgG1), 0.97 (IgG2), and 0.48 (IgG3), with sensitivities of 70.8, 87.5, 83.3, and 85.4%, and specificities of 70.8, 59.1, 77.3, and 56.8%, respectively. There was an additive interaction between serum anti-ENO1-P6 and anti-PDIA3 total IgGs on the development of miscarriage (RERI = 23.6, AP = 0.79, SI = 5.37). Conclusion: The highly expressed ENO1-P6Abs may be important risk factors for euthyroid TAI-related miscarriage. The serum levels of ENO1-P6Abs may become good predictive markers for pregnancy loss in euthyroid TAI females, especially its IgG2 subclass expression.


Assuntos
Aborto Espontâneo , Autoanticorpos , Proteínas de Ligação a DNA , Fosfopiruvato Hidratase , Doenças da Glândula Tireoide , Autoimunidade , Biomarcadores Tumorais/imunologia , Proteínas de Ligação a DNA/imunologia , Epitopos , Feminino , Humanos , Imunoglobulina G , Fosfopiruvato Hidratase/imunologia , Gravidez , Doenças da Glândula Tireoide/imunologia , Glândula Tireoide/fisiopatologia , Proteínas Supressoras de Tumor/imunologia
2.
Microb Pathog ; 166: 105539, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35447314

RESUMO

Sporotrichosis is a cosmopolitan mycosis caused by pathogenic species of Sporothrix genus, that in Brazil is often acquired by zoonotic transmission involved infected cats with S. brasiliensis. Previous studies showed that the Sporothrix spp. recombinant enolase (rSsEno), a multifunctional protein with immunogenic properties, could be a promising target for vaccination against sporotrichosis in cats. Nevertheless, the considerable sequence identity (62%) of SsEno with its feline counterpart is a great concern. Here, we report the identification in silico, chemical synthesis and biological validation of six peptides of SsEno with low sequence identity to its cat orthologue. All synthesized peptides exhibit B-cell epitopes on the molecular surface of SsEno and proved to be highly reactive with the serum of infected mice with S. brasiliensis and sera of cats with sporotrichosis. Interestingly, our study revealed that anti-peptide sera did not react with the recombinant enolase from Felis catus (cats, rFcEno), thus, may not trigger autoimmune response in these felines if used as a vaccine antigen. The immunization with peptide mixture (PeptMix) formulated with Freund adjuvant (FA), induced high levels of antigen-specific IgG, IgG1 and IgG2b antibodies that conferred protection upon passive transference in infected BALB/c mice with S. brasiliensis. We also observed, that the FA+PeptMix formulation induced a Th1/Th2/Th17 cytokine profile ex vivo, associated with protecting effect against the experimental sporotrichosis. Our results suggest that the six SsEno-derived peptides here evaluated, could be used as safe antigens for the development of vaccine strategies against feline sporotrichosis, whether prophylactic or therapeutic.


Assuntos
Vacinas Fúngicas , Fosfopiruvato Hidratase , Esporotricose , Animais , Brasil , Gatos , Epitopos , Vacinas Fúngicas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/imunologia , Sporothrix/enzimologia , Sporothrix/genética , Esporotricose/prevenção & controle
3.
JCI Insight ; 6(24)2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34935642

RESUMO

Fibroproliferative disorders such as systemic sclerosis (SSc) have no effective therapies and result in significant morbidity and mortality. We recently demonstrated that the C-terminal domain of endostatin, known as E4, prevented and reversed both dermal and pulmonary fibrosis. Our goal was to identify the mechanism by which E4 abrogates fibrosis and its cell surface binding partner(s). Our findings show that E4 activated the urokinase pathway and increased the urokinase plasminogen activator (uPA) to type 1 plasminogen activator inhibitor (PAI-1) ratio. In addition, E4 substantially increased MMP-1 and MMP-3 expression and activity. In vivo, E4 reversed bleomycin induction of PAI-1 and increased uPA activity. In patients with SSc, the uPA/PAI-1 ratio was decreased in both lung tissues and pulmonary fibroblasts compared with normal donors. Proteins bound to biotinylated-E4 were identified as enolase-1 (ENO) and uPA receptor (uPAR). The antifibrotic effects of E4 required uPAR. Further, ENO mediated the fibrotic effects of TGF-ß1 and exerted TGF-ß1-independent fibrotic effects. Our findings suggest that the antifibrotic effect of E4 is mediated, in part, by regulation of the urokinase pathway and induction of MMP-1 and MMP-3 levels and activity in a uPAR-dependent manner, thus promoting extracellular matrix degradation. Further, our findings identify a moonlighting function for the glycolytic enzyme ENO in fibrosis.


Assuntos
Proteínas E4 de Adenovirus/imunologia , Biomarcadores Tumorais/imunologia , Proteínas de Ligação a DNA/imunologia , Fibrose/imunologia , Fosfopiruvato Hidratase/imunologia , Proteínas Supressoras de Tumor/imunologia , Ativador de Plasminogênio Tipo Uroquinase/imunologia , Humanos , Transdução de Sinais , Transfecção
4.
J Immunol Res ; 2021: 6629824, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34222496

RESUMO

Haemophilus influenzae is a common organism of the human upper respiratory tract; this bacterium is responsible of a wide spectrum for respiratory infections and can generate invasive diseases such as meningitis and septicemia. These infections are associated with H. influenzae encapsulated serotype b. However, the incidence of invasive disease caused by nontypeable H. influenzae (NTHi) has increased in the post-H. influenzae serotype b (Hib) vaccine era. Currently, an effective vaccine against NTHi is not available; due to this, it is important to find an antigen capable to confer protection against NTHi infection. In this study, 10 linear B cell epitopes and 13 CTL epitopes and a putative plasminogen-binding motif (252FYNKENGMY260) and the presence of enolase on the surface of different strains of H. influenzae were identified in the enolase sequence of H. influenzae. Both in silico and experimental results showed that recombinant enolase from H. influenzae is immunogenic that could induce a humoral immune response; this was observed mediating the generation of specific polyclonal antibodies anti-rNTHiENO that recognize typeable and nontypeable H. influenzae strains. The immunogenic properties and the superficial localization of enolase in H. influenzae, important characteristics to be considered as a new candidate for the development of a vaccine, were demonstrated.


Assuntos
Proteínas de Bactérias/imunologia , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/imunologia , Haemophilus influenzae/imunologia , Fosfopiruvato Hidratase/imunologia , Infecções Respiratórias/prevenção & controle , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Clonagem Molecular , Biologia Computacional , Epitopos/genética , Epitopos/imunologia , Infecções por Haemophilus/imunologia , Infecções por Haemophilus/microbiologia , Vacinas Anti-Haemophilus/genética , Vacinas Anti-Haemophilus/uso terapêutico , Haemophilus influenzae/enzimologia , Haemophilus influenzae/genética , Humanos , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/isolamento & purificação , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Infecções Respiratórias/imunologia , Infecções Respiratórias/microbiologia , Vacinas de Subunidades/genética , Vacinas de Subunidades/imunologia , Vacinas de Subunidades/uso terapêutico
5.
Biomed Res Int ; 2021: 8845826, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34095312

RESUMO

Immunotherapy, a treatment based on host immune system activation, has been shown to provide a substitute for marginally effective conventional chemotherapy in controlling visceral leishmaniasis (VL), the deadliest form of leishmaniasis. As the majority of endemic inhabitants exhibit either subclinical or asymptomatic infection which often develops into the active disease state, therapeutic intervention seems to be an important avenue for combating infections by stimulating the natural defense system of infected individuals. With this perspective, the present study focuses on two immunodominant Leishmania (L.) donovani antigens (triosephosphate isomerase and enolase) previously proved to be potent prophylactic VL vaccine candidates, for generating a recombinant chimeric antigen. This is based on the premise that in a heterogeneous population, a multivalent antigen vaccine would be required for an effective response against leishmaniasis (a complex parasitic disease). The resulting molecule rLdT-E chimeric protein was evaluated for its immunogenicity and immunotherapeutic efficacy. A Th1 stimulating adjuvant BCG was employed with the protein which showed a remarkable 70% inhibition of splenic parasitic multiplication positively correlated with boosted Th1 dominant immune response against lethal L. donovani challenge in hamsters as evidenced by high IFN-γ and TNF-α and low IL-10. In addition, immunological analysis of antibody subclass presented IgG2-based humoral response besides considerable delayed-type hypersensitivity and lymphocyte proliferative responses in rLdT-E/BCG-treated animals. Our observations indicate the potential of the chimera towards its candidature for an effective vaccine against Leishmania donovani infection.


Assuntos
Leishmania donovani/genética , Leishmaniose Visceral/terapia , Células Th1/imunologia , Imunidade Adaptativa/imunologia , Animais , Antígenos de Protozoários/imunologia , Quimera , Cricetinae , Citocinas/metabolismo , Feminino , Fatores Imunológicos/metabolismo , Imunoterapia/métodos , Leishmania donovani/imunologia , Leishmania donovani/patogenicidade , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/metabolismo , Leucócitos Mononucleares/metabolismo , Ativação Linfocitária/imunologia , Linfócitos/metabolismo , Fosfopiruvato Hidratase/imunologia , Proteínas de Protozoários/metabolismo , Proteínas Recombinantes/metabolismo , Triose-Fosfato Isomerase/imunologia , Vacinas/farmacologia
6.
ACS Appl Mater Interfaces ; 13(24): 29179-29186, 2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34101420

RESUMO

This research put forward a novel split-type electrochemical (EC) immunosensor which integrated the controlled-release strategy with EC detection for application in the field of biosensing. Concretely, ascorbic acid (AA) was packaged in a cadmium sulfide (CdS)-capped spherical mesoporous bioactive glass (SBG) nanocarrier (SBGCdS) on account of encapsulation technology. To reduce the complexity of the bioanalysis, the detection antibody-labeled SBGCdS-AA bioconjugate was applied in a 96-well microplate for the immunoreaction process, which is independent of the EC determination procedure. Thus, the immune interference and steric hindrance caused by the accumulation of nanomaterials on the electrode could be minimized. Subsequently, AA was released efficiently via the destruction effect of dithiothreitol on the disulfide bond. In addition, for the as-prepared FcAI/l-Cys/gold nanoparticles (GNPs)/porous BiVO4 (p-BVO)/ITO EC sensing platform in the detection solution, the synergetic catalysis of Fc and GNPs/p-BVO toward the oxidation of the released AA could be realized, which triggered AA-mediated significant signal magnification throughout this study. In particular, p-BVO with an ordered nanoarray structure could accelerate the electron transfer to assist in sensitivity improvement of this system. This novel biosensor was capable of assaying the neuron-specific enolase (NSE) biomarker sensitively, from which a linear range of 0.001-100 ng/mL was derived along with a low detection limit of 1.08 pg/mL. An innovative way could be paved in the bioanalysis of NSE and other biomarkers.


Assuntos
Ácido Ascórbico/química , Imunoensaio/métodos , Nanopartículas Metálicas/química , Fosfopiruvato Hidratase/sangue , Anticorpos Imobilizados/imunologia , Biomarcadores/sangue , Técnicas Biossensoriais/métodos , Bismuto/química , Compostos de Cádmio/química , Técnicas Eletroquímicas , Ouro/química , Humanos , Limite de Detecção , Fosfopiruvato Hidratase/imunologia , Porosidade , Sulfetos/química , Vanadatos/química
7.
J Leukoc Biol ; 109(3): 633-644, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-32810329

RESUMO

The pathogenesis of ulcerative colitis (UC) is to be further investigated. House dust mites (HDM) are highly associated with the pathogenesis of immune inflammation in the body. This study aims to investigate the role of enolase (one of the HDM-derived proteins)-specific cross Abs in the induction of UC-like inflammation. The enolase specific IgG (EsIgG) was identified in UC patients by mass spectrometry. Mice were treated with EsIgG to induce inflammation in the colon mucosa. EsIgG was detected in the serum and the colon tissues of UC patients, which was positively correlated with the polymorphonuclear neutrophil (PMN) counts in the blood and colon tissues of UC patients. EsIgG formed immune complexes with the constitutive enolase in the UC colon epithelium that activated complement, induced epithelial cell apoptosis, compromised epithelial barrier functions, and resulted in UC-like inflammation in the mouse colon. In summary, UC patients have high serum levels of Abs against HDM-derived enolase and intestinal epithelial cell-derived enolase. These Abs attack the colonic epithelium to induce UC-like inflammation.


Assuntos
Anticorpos/metabolismo , Inflamação/patologia , Intestinos/patologia , Neutrófilos/patologia , Fosfopiruvato Hidratase/imunologia , Adulto , Animais , Apoptose , Colite Ulcerativa/sangue , Colite Ulcerativa/imunologia , Colo/patologia , Ativação do Complemento/imunologia , Células Epiteliais/metabolismo , Feminino , Humanos , Imunoglobulina G/sangue , Inflamação/sangue , Inflamação/imunologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Pyroglyphidae/imunologia
8.
Clin Exp Med ; 21(1): 73-78, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32857322

RESUMO

The objective of the study was to investigate the value of anti-α-enolase antibody (Ab) combined with RDW in evaluating the activity of systemic lupus erythematosus (SLE). Levels of serum anti-α-enolase Ab and RDW were detected in 193 SLE patients and 98 healthy controls by ELISA and automatic blood cell counter (XN9000), respectively. Furthermore, the correlation between anti-α-enolase Ab and RDW in evaluating the activity of SLE was evaluated by correlation analysis. The level of anti-α-enolase Ab (9.16 ± 0.44 ng/mL in stable group and 10.26 ± 0.36 ng/mL in activity group) was significantly higher than that in the healthy control (7.05 ± 0.27 ng/mL). The level of RDW (12.92% ± 1.23% in stable group and 13.57% ± 2.12% in activity group) was significantly higher than that in the healthy control (12.46% ± 0.61%). The levels of anti-α-enolase Ab or RDW in SLE patients were positively correlated with SLEDAI-2 K score (r= 0.75, r = 0.73), respectively. Compared with the anti-α-enolase Ab (AUC: 78.0%) or RDW (AUC:80.0%) alone, anti-α-enolase Ab combined with RDW (AUC: 81.0%) had the best of the effectiveness of evaluating activity of SLE. These data suggested that combined anti-α-enolase Ab with RDW might be good biomarker to predict the activity of SLE in clinical.


Assuntos
Anticorpos Antinucleares/imunologia , Biomarcadores/sangue , Índices de Eritrócitos , Eritrócitos/química , Lúpus Eritematoso Sistêmico/patologia , Fosfopiruvato Hidratase/imunologia , Adulto , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Prognóstico , Estudos Retrospectivos
9.
Rheumatology (Oxford) ; 60(7): 3176-3188, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-33374003

RESUMO

OBJECTIVES: Serum anti-dsDNA and anti-nucleosome IgGs have been proposed as signatures for SLE and LN in limited numbers of patients. We sought to show higher sensitivity and specificity of the same antibodies with the IgG2 isotype and included IgG2 antibodies vs specific intracellular antigens in the analysis. METHODS: A total of 1052 SLE patients with (n = 479) and without (n = 573) LN, recruited at different times from the beginning of symptoms, were included in the study. Patients with primary APS (PAPS, n = 24), RA (RA, n = 24) and UCTD (UCTD, n = 96) were analysed for comparison. Anti-nucleosome (dsDNA, Histone2A, Histone3), anti-intracellular antigens (ENO1), anti-annexin A1 and anti-C1q IgG2 were determined by non-commercial techniques. RESULTS: The presence in the serum of the IgG2 panel was highly discriminatory for SLE/LN vs healthy subjects. Serum levels of anti-dsDNA and anti-C1q IgG2 were more sensitive than those of IgGs (Farr radioimmunoassay/commercial assays) in identifying SLE patients at low-medium increments. Of more importance, serum positivity for anti-ENO1 and anti-H2A IgG2 discriminated between LN and SLE (ROC T0-12 months), and high levels at T0-1 month were detected in 63% and 67%, respectively, of LN, vs 3% and 3%, respectively, of SLE patients; serum positivity for each of these was correlated with high SLEDAI values. Minor differences existed between LN/SLE and the other rheumatologic conditions. CONCLUSION: Nephritogenic IgG2 antibodies represent a specific signature of SLE/LN, with a few overlaps with other rheumatologic conditions. High levels of anti-ENO1 and anti-H2A IgG2 correlated with SLE activity indexes and were discriminatory between SLE patients limited to the renal complication and other SLE patients. TRIAL REGISTRATION: The Zeus study was registered at https://clinicaltrials.gov, NCT02403115.


Assuntos
Anticorpos Antinucleares/imunologia , Imunoglobulina G/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Nefrite Lúpica/imunologia , Adolescente , Adulto , Anexina A1/imunologia , Especificidade de Anticorpos , Síndrome Antifosfolipídica/imunologia , Artrite Reumatoide/imunologia , Biomarcadores Tumorais/imunologia , Complemento C1q/imunologia , Estudos Transversais , DNA/imunologia , Proteínas de Ligação a DNA/imunologia , Feminino , Histonas/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Nucleossomos/imunologia , Fosfopiruvato Hidratase/imunologia , Proteínas Supressoras de Tumor/imunologia , Doenças do Tecido Conjuntivo Indiferenciado/imunologia , Adulto Jovem
10.
Rheumatology (Oxford) ; 60(7): 3388-3397, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-33351137

RESUMO

OBJECTIVES: Circulating anti-ENO1 and anti-H2A IgG2 have been identified as specific signatures of LN in a cross-over approach. We sought to show whether the same antibodies identify selected population of patients with LN with potentially different clinical outcomes. METHODS: Here we report the prospective analysis over 36 months of circulating IgG2 levels in patients with newly diagnosed LN (n=91) and SLE (n=31) and in other patients with SLE recruited within 2 years from diagnosis (n=99). Anti-podocyte (ENO1), anti-nucleosome (DNA, histone 2 A, histone 3) and anti-circulating proteins (C1q, AnnexinA1-ANXA1) IgG2 antibodies were determined by home-made techniques. RESULTS: LN patients were the main focus of the study. Anti-ENO1, anti-H2A and anti-ANXA1 IgG2 decreased in parallel to proteinuria and normalized within 12 months in the majority of patients while anti-dsDNA IgG2 remained high over the 36 months. Anti-ENO1 and anti-H2A had the highest association with proteinuria (Heat Map) and identified the highest number of patients with high proteinuria (68% and 71% respectively) and/or with reduced estimated glomerula filtration rate (eGFR) (58% for both antibodies) compared with 23% and 17% of anti-dsDNA (agreement analysis). Anti-ENO1 positive LN patients had higher proteinuria than negative patients at T0 and presented the maximal decrement within 12 months. CONCLUSIONS: Anti-ENO1, anti-H2A and anti-ANXA1 antibodies were associated with high proteinuria in LN patients and Anti-ENO1 also presented the maximal reduction within 12 months that paralleled the decrease of proteinuria. Anti-dsDNA were not associated with renal outcome parameters. New IgG2 antibody signatures should be utilized as tracers of personalized therapies in LN. TRIAL REGISTRATION: The Zeus study was registered at https://clinicaltrials.gov (study number: NCT02403115).


Assuntos
Imunoglobulina G/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Nefrite Lúpica/imunologia , Adulto , Anexina A1/imunologia , Anticorpos Antinucleares/imunologia , Autoanticorpos/imunologia , Biomarcadores Tumorais/imunologia , Complemento C1q/imunologia , DNA/imunologia , Proteínas de Ligação a DNA/imunologia , Progressão da Doença , Feminino , Histonas/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Nucleossomos/imunologia , Fosfopiruvato Hidratase/imunologia , Estudos Prospectivos , Proteínas Supressoras de Tumor/imunologia
11.
Clin J Am Soc Nephrol ; 15(12): 1762-1776, 2020 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-33257410

RESUMO

BACKGROUND AND OBJECTIVES: Patients with membranous nephropathy can have circulating autoantibodies against membrane-bound (phospholipase A2 receptor 1 [PLA2R1] and thrombospondin type-1 domain containing 7A [THSD7A]) and intracellular (aldose reductase, SOD2, and α-enolase) podocyte autoantigens. We studied their combined association with clinical outcomes. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Serum levels of anti-PLA2R1, anti-THSD7A, anti-aldose reductase, anti-SOD2, and anti-α-enolase autoantibodies were determined in 285 patients at diagnosis and during follow-up using standardized and homemade assays. An eGFR>60 ml/min per 1.73 m2 and remission of proteinuria (<0.3/<3.5 g per d) after 12 months were the outcomes of interest. RESULTS: At diagnosis, 182 (64%), eight (3%), and 95 (33%) patients were anti-PLA2R1+, anti-THSD7A+, and double negative, respectively. The prevalence of a detectable antibody to at least one intracellular antigen was similarly distributed in patients who were anti-PLA2R1+ (n=118, 65%) and double negative (n=64, 67%). Positivity for anti-PLA2R1, anti-SOD2, and anti-α-enolase antibodies and higher titers at diagnosis were associated with poor clinical outcome independently to each other. Combined positivity for anti-PLA2R1, anti-SOD2, and anti-α-enolase was associated with highest risk of poor outcome (odds ratio, 5.5; 95% confidence interval, 1.2 to 24; P=0.01). In Kaplan-Meier analysis, patients who were anti-PLA2R1+/anti-SOD2+ or anti-PLA2R1+/anti-α-enolase+ had lower eGFR at 12 months compared with patients who were anti-PLA2R1+/anti-SOD2- or anti-α-enolase-. Predictive tests (net reclassification index and area under the curve-receiver-operating characteristic analysis) showed that combined assessment of antibodies improved classification of outcome in 22%-34% of cases for partial remission of proteinuria and maintenance of normal eGFR. For patients with nephrotic syndrome at diagnosis, anti-SOD2 positivity and high anti-PLA2R1 titer were associated with a lack of complete remission. Patients who were anti-PLA2R1-/anti-intracellular antigens- had the lowest proteinuria and the highest eGFR at diagnosis and the lowest risk of lower eGFR at 12 months. Epitope spreading was present in 81% of patients who were anti-PLA2R1+ and was associated with increased positivity for intracellular antigens and poor eGFR at diagnosis and 12 months. CONCLUSIONS: Combined serological analysis of autoantibodies targeting membrane-bound and intracellular autoantigens identifies patients with poor clinical outcomes.


Assuntos
Aldeído Redutase/imunologia , Autoanticorpos/sangue , Biomarcadores Tumorais/imunologia , Proteínas de Ligação a DNA/imunologia , Glomerulonefrite Membranosa/imunologia , Fosfopiruvato Hidratase/imunologia , Receptores da Fosfolipase A2/imunologia , Superóxido Dismutase/imunologia , Trombospondinas/imunologia , Proteínas Supressoras de Tumor/imunologia , Adulto , Idoso , Biomarcadores/sangue , Estudos Transversais , Feminino , França , Glomerulonefrite Membranosa/sangue , Glomerulonefrite Membranosa/diagnóstico , Glomerulonefrite Membranosa/terapia , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Estudos Retrospectivos , Testes Sorológicos , Fatores de Tempo
12.
Int J Mol Sci ; 21(21)2020 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-33105833

RESUMO

Significant amounts of enolase-a cytosolic enzyme involved in the glycolysis pathway-are exposed on the cell surface of Candida yeast. It has been hypothesized that this exposed enolase form contributes to infection-related phenomena such as fungal adhesion to human tissues, and the activation of fibrinolysis and extracellular matrix degradation. The aim of the present study was to characterize, in structural terms, the protein-protein interactions underlying these moonlighting functions of enolase. The tight binding of human vitronectin, fibronectin and plasminogen by purified C. albicans and C. tropicalis enolases was quantitatively analyzed by surface plasmon resonance measurements, and the dissociation constants of the formed complexes were determined to be in the 10-7-10-8 M range. In contrast, the binding of human proteins by the S.cerevisiae enzyme was much weaker. The chemical cross-linking method was used to map the sites on enolase molecules that come into direct contact with human proteins. An internal motif 235DKAGYKGKVGIAMDVASSEFYKDGK259 in C. albicans enolase was suggested to contribute to the binding of all three human proteins tested. Models for these interactions were developed and revealed the sites on the enolase molecule that bind human proteins, extensively overlap for these ligands, and are well-separated from the catalytic activity center.


Assuntos
Fibronectinas/metabolismo , Fosfopiruvato Hidratase/metabolismo , Plasminogênio/metabolismo , Vitronectina/metabolismo , Motivos de Aminoácidos , Anticorpos/metabolismo , Ligação Competitiva , Candida albicans/enzimologia , Candida tropicalis/enzimologia , Citosol/enzimologia , Fibronectinas/química , Interações Hospedeiro-Patógeno/fisiologia , Humanos , Proteínas Imobilizadas/metabolismo , Modelos Moleculares , Fosfopiruvato Hidratase/química , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/imunologia , Plasminogênio/química , Vitronectina/química
13.
Mikrochim Acta ; 187(8): 480, 2020 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-32743701

RESUMO

MoS2 nanosheets were prepared by exfoliating MoS2 bulk crystals with ultrasonication in N-methylpyrrolidone and were integrated with gold nanostars (AuNS) to fabricate an AuNS/MoS2 nanocomposite. All nanomaterials were characterized by transmission electron microscope, scanning electron microscope, ultraviolet-visible spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. AuNS/MoS2 nanocomposites were coated onto a glassy carbon electrode (GCE) surface to construct a nanointerface for immobilizing neuron-specific enolase antibody (anti-NSE) thus forming a photoelectrochemical immunoassay system. AuNS can significantly promote the photoelectric conversion of MoS2 nanosheets improving the performance for a photoelectrochemical assay. Being illuminated with white light LED and controlling the potential at 0.05 V (vs. SCE), the photocurrent generated from anti-NSE(BSA)/AuNS/MoS2/GCE using 0.15 mol L-1 ascorbic acid as electron donor can be recorded with amperometry and used as an output signal for NSE quantitative assay. Under optimized experimental conditions, the photocurrent variation for the affinity-binding NSE is proportional to the logarithm of NSE concentration in the range 5.0 pg mL-1   to 1.5 ng mL-1 with a detection limit of 3.5 pg mL-1 (S/N = 3). The practicability of the PEC immunoassay system was evaluated by determining NSE in clinical serum samples. The recoveries ranged from 93.0 to 103% for the determination of NSE in serum samples with a standard addition method. The PEC immunoassay system possesses good accuracy for determining NSE in real samples. Graphical abstract.


Assuntos
Dissulfetos/química , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Nanopartículas Metálicas/química , Molibdênio/química , Fosfopiruvato Hidratase/sangue , Anticorpos Imobilizados/imunologia , Dissulfetos/efeitos da radiação , Ouro/química , Humanos , Luz , Nanopartículas Metálicas/efeitos da radiação , Molibdênio/efeitos da radiação , Fosfopiruvato Hidratase/imunologia , Processos Fotoquímicos
14.
J Immunother Cancer ; 8(1)2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32561639

RESUMO

BACKGROUND: Stress-induced post-translational modifications occur during autophagy and can result in generation of new epitopes and immune recognition. One such modification is the conversion of arginine to citrulline by peptidylarginine deiminase enzymes. METHODS: We used Human leukocyte antigen (HLA) transgenic mouse models to assess the immunogenicity of citrullinated peptide vaccine by cytokine Enzyme linked immunosorbant spot (ELISpot) assay. Vaccine efficacy was assessed in tumor therapy studies using HLA-matched B16 melanoma and ID8 ovarian models expressing either constitutive or interferon-gamma (IFNγ) inducible Major Histocompatibility Complex (MHC) class II (MHC-II) as represented by most human tumors. To determine the importance of CD4 T cells in tumor therapy, we analyzed the immune cell infiltrate into murine tumors using flow cytometry and performed therapy studies in the presence of CD4 and CD8 T cell depletion. We assessed the T cell repertoire to citrullinated peptides in ovarian cancer patients and healthy donors using flow cytometry. RESULTS: The combination of citrullinated vimentin and enolase peptides (Modi-1) stimulated strong CD4 T cell responses in mice. Responses resulted in a potent anti-tumor therapy against established tumors and generated immunological memory which protected against tumor rechallenge. Depletion of CD4, but not CD8 T cells, abrogated the primary anti-tumor response as well as the memory response to tumor rechallenge. This was further reinforced by successful tumor regression being associated with an increase in tumor-infiltrating CD4 T cells and a reduction in tumor-associated myeloid suppressor cells. The anti-tumor response also relied on direct CD4 T cell recognition as only tumors expressing MHC-II were rejected. A comparison of different Toll-like receptor (TLR)-stimulating adjuvants showed that Modi-1 induced strong Th1 responses when combined with granulocyte-macrophage colony-stimulating factor (GMCSF), TLR9/TLR4, TLR9, TLR3, TLR1/2 and TLR7 agonists. Direct linkage of the TLR1/2 agonist to the peptides allowed the vaccine dose to be reduced by 10-fold to 100-fold without loss of anti-tumor activity. Furthermore, a CD4 Th1 response to the citrullinated peptides was seen in ovarian cancer patients. CONCLUSIONS: Modi-1 citrullinated peptide vaccine induces potent CD4-mediated anti-tumor responses in mouse models and a CD4 T cell repertoire is present in ovarian cancer patients to the citrullinated peptides suggesting that Modi-1 could be an effective vaccine for ovarian cancer patients.


Assuntos
Vacinas Anticâncer/imunologia , Melanoma Experimental/terapia , Fosfopiruvato Hidratase/imunologia , Vimentina/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/genética , Linhagem Celular Tumoral , Citrulinação/imunologia , Feminino , Antígenos HLA/genética , Antígenos HLA/imunologia , Humanos , Imunogenicidade da Vacina , Interferon gama/imunologia , Depleção Linfocítica , Masculino , Melanoma Experimental/imunologia , Camundongos , Camundongos Transgênicos , Fosfopiruvato Hidratase/genética , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/genética , Vacinas Combinadas/imunologia , Vacinas de Subunidades/administração & dosagem , Vacinas de Subunidades/genética , Vacinas de Subunidades/imunologia , Vimentina/genética
15.
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
16.
Int Immunopharmacol ; 85: 106563, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32442899

RESUMO

Alpha-enolase (ENO1) is a ubiquitous protein. Patients with autoimmune thyroiditis-associated encephalopathy have high serum ENO1Ab titers. We aimed to explore whether ENO1Ab was the pathogenic antibody in the thyroid and brain. The serum ENO1Ab titers were significantly increased in the mice immunized with Thyroglobulin (Tg). And in the mice immunized with ENO1, serum levels of both TgAb and thyroid-stimulating hormone (TSH) were significantly increased. Obvious CD16+ cell infiltration, IgG deposit and cleaved caspase-3 were observed in the thyroid of ENO1-immunized mice. Spatial learning and memory abilities and synaptic functions were impaired in ENO1-immunized mice. Furthermore, the expression levels of Iba-1, GFAP, interlukin-6, CDK5, and phosphorylated tau were increased, and endothelial tight junction proteins were decreased in the brain of ENO1-immunized mice. These results suggest that ENO1Ab can cause thyrocyte damage via ADCC effect and impair cerebral function by disrupting the blood-brain barrier.


Assuntos
Anticorpos/imunologia , Autoantígenos/imunologia , Encefalopatias/imunologia , Encéfalo/imunologia , Fosfopiruvato Hidratase/imunologia , Glândula Tireoide/imunologia , Tireoidite Autoimune/imunologia , Animais , Anticorpos/sangue , Encéfalo/irrigação sanguínea , Encefalopatias/sangue , Citocinas/genética , Feminino , Aprendizagem em Labirinto , Camundongos Endogâmicos CBA , Microvasos , Fosfopiruvato Hidratase/sangue , Baço/imunologia , Tireoglobulina/imunologia , Tireoidite Autoimune/sangue , Tireotropina/sangue , Tiroxina/sangue
17.
BMC Ophthalmol ; 20(1): 132, 2020 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-32252709

RESUMO

BACKGROUND: Evidence-based criteria for the treatment of autoimmune retinopathy (AIR) have not been established. The pathology and clinical features of each antibody causing AIR, and its long-term course are still undetermined. We report our findings in a case of non-paraneoplastic AIR (npAIR) that developed in the fellow eye 10 years after the onset in the first eye. CASE PRESENTATION: Our patient had photophobia in both eyes and a rapidly progressing visual field defect in his right eye at the initial examination. He was diagnosed with non-paraneoplastic AIR based on the clinical findings and immunoblot analyses for anti-retinal antibodies, and he was treated with steroids. Ten years later, a visual field defect developed in the fellow eye, and a diagnosis of npAIR was made. Immunoblot analyses were positive for anti-α-enolase antibodies. He was treated with steroids, immunosuppressants, and plasma exchange. However, the response to the treatment was poor and both eyes eventually became blind. CONCLUSIONS: As best we know, this is the first case report of npAIR that developed in the fellow eye over 10 years after the development in the first eye. Long-term follow-up and a search for tumor lesions are necessary in cases of npAIR. Further understanding of the long-term course of AIR can contribute to an understanding of the pathology and treatment of npAIR.


Assuntos
Doenças Autoimunes/etiologia , Síndromes Paraneoplásicas Oculares/etiologia , Doenças Retinianas/etiologia , Autoanticorpos/sangue , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/terapia , Cegueira/etiologia , Eletrorretinografia , Angiofluoresceinografia , Glucocorticoides/uso terapêutico , Humanos , Immunoblotting , Imunossupressores/uso terapêutico , Masculino , Pessoa de Meia-Idade , Síndromes Paraneoplásicas Oculares/diagnóstico , Síndromes Paraneoplásicas Oculares/terapia , Fosfopiruvato Hidratase/imunologia , Troca Plasmática , Doenças Retinianas/diagnóstico , Doenças Retinianas/terapia , Fatores de Tempo , Tomografia de Coerência Óptica , Acuidade Visual , Testes de Campo Visual
18.
Mol Neurobiol ; 57(5): 2265-2278, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32006234

RESUMO

Suppression of ubiquitin proteasome pathway (UPP) and stimulation of caspase-3 are involved in neurodegeneration. Can UPP activators and caspase-3 inhibitors ameliorate neurodegeneration? Here, we found a novel neuronal cell death accompanied with UPP activation and caspase-3 inhibition. Recently, plasmalemmal neuron-specific enolase (NSE) has been identified as one of membrane targets of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2). 15d-PGJ2 induces neuronal apoptosis via activating caspase-3 and inactivating UPP, whereas the anti-NSE antibody inactivated caspase-3, activated UPP, and caused neuronal cell death. The anti-NSE antibody activated caspase-1 (pyroptosis marker), but not condense chromatin (apoptosis marker). The anti-NSE antibody declined intracellular level of ATP, which is not altered in pyroptosis. The intracellular level of calcium is elevated in necrosis and pyroptosis, but its chelator did not ameliorate the neurotoxicity of anti-NSE. Thiol antioxidants such as N-acetyl cysteine and glutathione reduced the neurotoxicity of 15d-PGJ2 but enhanced that of the anti-NSE antibody. The anti-NSE antibody incorporated propidium iodide into neurons through the disrupted plasma membrane, which are not observed in ferroptosis and autophagic cell death. Thus, the anti-NSE antibody induced neuronal cell death in a novel fashion distinguished from necrosis, necroptosis, apoptosis, pyroptosis, ferroptosis, and autophagic cell death.


Assuntos
Caspase 3/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Imunoglobulina G/farmacologia , Neurônios/efeitos dos fármacos , Fosfopiruvato Hidratase/imunologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Ubiquitina/metabolismo , Acetilcisteína/farmacologia , Trifosfato de Adenosina/metabolismo , Animais , Antioxidantes/farmacologia , Sinalização do Cálcio , Caspase 1/metabolismo , Córtex Cerebral/citologia , Cromatina/ultraestrutura , Ativação Enzimática/efeitos dos fármacos , Feminino , Glutationa/farmacologia , Cabras/imunologia , Proteínas de Choque Térmico HSP70/metabolismo , Imunoglobulina G/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Neuritos/efeitos dos fármacos , Neurônios/citologia , Fosfopiruvato Hidratase/fisiologia , Gravidez , Prostaglandina D2/análogos & derivados , Prostaglandina D2/fisiologia , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Coelhos/imunologia , Ratos , Ratos Wistar , Especificidade da Espécie , Ubiquitinação/efeitos dos fármacos
19.
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
20.
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
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