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2.
Fish Shellfish Immunol ; 93: 313-321, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31351111

RESUMO

The Src family kinases (SFK) are involved in signaling transductions that regulate numerous biological activities including host-virus interaction. These features of SFK have been well explored in vertebrates, however, in shrimp, the invertebrate SFK family member Src64B, has not been characterized and therefore its role in shrimp-virus interaction remains unknown. In this study, two Litopenaeus vannamei Src64B isoforms (designated LvSrc64B1 and LvSrc64B2) were first cloned and their role in white spot syndrome virus (WSSV) infection was explored. Bioinformatics analysis revealed that LvSrc64B1 and LvSrc64B2 were similar to other Src64B family members, with high homology in primary and tertiary structures, and contained the conserved SFK functional domains, as well as the putative myristylation and phosphorylation sites. Tissue distribution analysis showed that both LvSrc64B isoforms were ubiquitously expressed, albeit distinctively in the tested tissues. In addition, transcript levels of LvSrc64B1 and LvSrc64B2 were significantly induced following WSSV challenge and had similar expression patterns. Furthermore, siRNA-mediated knockdown of LvSrc64B1 and LvSrc64B2 followed by WSSV infection resulted in increased expression of viral genes, enhanced viral DNA replication, and elevation of hemocytes apoptosis. Depletion of LvSrc64B1 and LvSrc64B2 also reduced shrimp survival upon WSSV infection. In conclusion, the current data strongly suggest that Src64B is a host factor that inhibits WSSV replication by modulating apoptosis in shrimp.


Assuntos
Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Biologia Computacional , Perfilação da Expressão Gênica , Filogenia , Proteínas Tirosina Quinases/química , Alinhamento de Sequência , Replicação Viral
3.
BMC Res Notes ; 12(1): 234, 2019 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-31010428

RESUMO

OBJECTIVE: We recently showed that the anti-helminthic compound mebendazole (MBZ) has immunomodulating activity by inducing a M2 to M1 phenotype switch in monocyte/macrophage models. In the present study we investigated the potential role of protein kinases in mediating this effect. RESULTS: MBZ potently binds and inhibits Dual specificity tyrosine-phosphorylation-regulated kinase 1B (DYRK1B) with a Kd and an IC50 of 7 and 360 nM, respectively. The specific DYRK1B inhibitor AZ191 did not mimic the cytokine release profile of MBZ in untreated THP-1 monocytes. However, in THP-1 cells differentiated into macrophages, AZ191 strongly induced a pro-inflammatory cytokine release pattern similar to MBZ and LPS/IFNγ. Furthermore, like MBZ, AZ191 increased the expression of the M1 marker CD80 and decreased the M2 marker CD163 in THP-1 macrophages. In this model, AZ191 also increased phospho-ERK activity although to a lesser extent compared to MBZ. Taken together, the results demonstrate that DYRK1B inhibition could, at least partly, recapitulate immune responses induced by MBZ. Hence, DYRK1B inhibition induced by MBZ may be part of the mechanism of action to switch M2 to M1 macrophages.


Assuntos
Antinematódeos/farmacologia , Macrófagos/efeitos dos fármacos , Mebendazol/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/antagonistas & inibidores , Antígenos CD/genética , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/imunologia , Antinematódeos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica , Compostos Heterocíclicos com 2 Anéis/farmacologia , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interferon gama/farmacologia , Interleucinas/genética , Interleucinas/imunologia , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/imunologia , Mebendazol/metabolismo , Fosforilação/efeitos dos fármacos , Ligação Proteica , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Proteínas Proto-Oncogênicas c-abl/genética , Proteínas Proto-Oncogênicas c-abl/imunologia , Pirimidinas/farmacologia , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Transdução de Sinais , Células THP-1 , Acetato de Tetradecanoilforbol/farmacologia
4.
PLoS One ; 14(3): e0214310, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30901349

RESUMO

Ocular toxoplasmosis (OT), mostly retinochorioditis, is a major feature of infection with the protozoan parasite Toxoplasma gondii. The pathophysiology of this infection is still largely elusive; especially mouse models are not yet well developed. In contrast, numerous in vitro studies showed the highly Toxoplasma strain dependent nature of the host-parasite interactions. Some distinct polymorphic virulence factors were characterized, notably the rhoptry protein ROP16. Here, we studied the strain-dependent pathophysiology in our OT mouse model. Besides of two wild type strains of the canonical I (RH, virulent) and II (PRU, avirulent) types, we used genetically engineered parasites, RHΔROP16 and PRU ROP16-I, expressing the type I allele of this virulence factor. We analyzed retinal integrity, parasite proliferation and retinal expression of cytokines. PRU parasites behaved much more virulently in the presence of a type I ROP16. In contrast, knockout of ROP16 in the RH strain led to a decrease of intraocular proliferation, but no difference in retinal pathology. Cytokine quantification in aqueous humor showed strong production of Th1 and inflammatory markers following infection with the two strains containing the ROP16-I allele. In strong contrast, immunofluorescence images showed that actual expression of most cytokines in retinal cells is rapidly suppressed by type I strain infection, with or without the involvement of its homologous ROP16 allele. This demonstrates the particular immune privileged situation of the retina, which is also revealed by the fact that parasite proliferation is nearly exclusively observed outside the retina. In summary, we further developed a promising OT mouse model and demonstrated the specific pathology in retinal tissues.


Assuntos
Citocinas/metabolismo , Proteínas Tirosina Quinases/imunologia , Proteínas de Protozoários/imunologia , Toxoplasma/patogenicidade , Toxoplasmose Ocular/parasitologia , Animais , Modelos Animais de Doenças , Feminino , Engenharia Genética , Camundongos , Proteínas Tirosina Quinases/genética , Proteínas de Protozoários/genética , Retina/imunologia , Retina/parasitologia , Toxoplasma/classificação , Toxoplasma/imunologia , Toxoplasmose Ocular/imunologia , Virulência
5.
J Cancer Res Clin Oncol ; 145(2): 281-291, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30656409

RESUMO

PURPOSE: Esophageal cancer is a common disease in China with low survival rate due to no obvious early symptoms and lack of effective screening strategies. Traditional treatments usually do not produce desirable results in patients with advanced esophageal cancer, so immunotherapy which relies on tumor-related antigens is needed to combat low survival rates effectively. Cancer-testis antigens (CTA), a large family of tumor-related antigens, have a strong in vivo immunogenicity and tumor-restricted expressing patterns in normal adult tissues. These two characteristics are ideal features of anticancer immunotherapy targets and, therefore, promoted the development of some studies of CTA-based therapy. To provide ideas for the role of the cancer-testis antigens MAGE-A, NY-ESO-1, LAGE-1, and TTK in esophageal cancer, we summarized their expression, prognostic value, and development in immunotherapy. METHODS: The relevant literature from PubMed is reviewed in this study. RESULTS: In esophageal cancer, although the relationship between expression of MAGE-A, NY-ESO-1, LAGE-1, and TTK and prognosis value is still in a controversial situation, MAGE-A, NY-ESO-1, LAGE-1, and TTK are highly expressed and can induce specific CTL cells to produce particular killing effect on tumor cells, and some clinical trials have demonstrated that immunotherapy for esophageal cancer patients is effective and safe, which provides a new therapeutic strategy for the treatment of esophageal cancer in the future. CONCLUSION: In this review, we summarize expression and prognostic value of MAGE-A, NY-ESO-1, LAGE-1, and TTK in esophageal cancer and point out recent advances in immunotherapy about them.


Assuntos
Antígenos de Neoplasias/imunologia , Neoplasias Esofágicas/imunologia , Neoplasias Esofágicas/terapia , Imunoterapia/métodos , Testículo/imunologia , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Antígenos de Superfície/genética , Antígenos de Superfície/imunologia , Antígenos de Superfície/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/imunologia , Proteínas de Ciclo Celular/metabolismo , Progressão da Doença , Neoplasias Esofágicas/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/imunologia , Humanos , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Prognóstico , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Proteínas Tirosina Quinases/metabolismo
6.
Biochim Biophys Acta Proteins Proteom ; 1867(1): 71-75, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29753089

RESUMO

Protein kinases are known to be implicated in various biological phenomena and diseases through their involvement in protein phosphorylation. Therefore, analysis of the activity of protein kinases by examination of their phosphorylation state is important to elucidate their mechanisms. However, a method for analyzing the phosphorylation state of entire protein kinases in cells is not established. In the present study, we developed a new profiling method to analyze the expression and phosphorylation state of protein kinases using a Multi-PK antibody and Phos-tag 2D-PAGE. When HL-60 cells were differentiated into macrophage-like cells induced by 12-O-tetradecanoylphorbol-13-acetate, we observed significant changes in the expression and phosphorylation state of immunoreactive spots by this method. These results show that tyrosine kinase expression levels and phosphorylation state are changed by differentiation. Taken together, the developed method will be a useful tool for analysis of intracellular tyrosine protein kinases.


Assuntos
Eletroforese em Gel Bidimensional/métodos , Proteínas Tirosina Quinases/análise , Proteínas Tirosina Quinases/metabolismo , Anticorpos Monoclonais , Eletroforese em Gel de Poliacrilamida/métodos , Células HL-60 , Humanos , Fosforilação/fisiologia , Proteínas Quinases/metabolismo , Proteínas Tirosina Quinases/imunologia , Proteômica/métodos
7.
J Immunol ; 201(9): 2602-2611, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30249810

RESUMO

Resolution of the inflammatory response requires coordinated regulation of pro- and anti-inflammatory mediator production, together with clearance of recruited inflammatory cells. Many different receptors have been implicated in phagocytosis of apoptotic cells (efferocytosis), including Mer, a receptor tyrosine kinase that can mediate recognition and subsequent internalization of apoptotic cells. In this manuscript, we examine the expression and function of the Tyro3/Axl/Mer (TAM) family of receptors by human monocytes. We demonstrate that the Mer ligand, protein S, binds to the surface of viable monocytes via phosphatidylserine-dependent and -independent mechanisms. Importantly, we have identified a novel role for receptor tyrosine kinase signaling in the augmentation of monocyte cytokine release in response to LPS. We propose that low-level phosphatidylserine exposure on the plasma membrane of viable monocytes allows protein S binding that leads to TAM-dependent augmentation of proinflammatory cytokine production. Our findings identify a potentially important role for TAM-mediated signaling during the initiation phase of inflammation.


Assuntos
Inflamação/imunologia , Monócitos/imunologia , Receptores Proteína Tirosina Quinases/imunologia , Humanos , Inflamação/metabolismo , Lipopolissacarídeos/imunologia , Monócitos/metabolismo , Proteína S/imunologia , Proteína S/metabolismo , Proteínas Tirosina Quinases/imunologia , Proteínas Tirosina Quinases/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , c-Mer Tirosina Quinase/imunologia , c-Mer Tirosina Quinase/metabolismo
8.
Front Immunol ; 9: 979, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29867957

RESUMO

Patients with primary immunodeficiency can be prone to severe Epstein-Barr virus (EBV) associated immune dysregulation. Individuals with mutations in the interleukin-2-inducible T-cell kinase (ITK) gene experience Hodgkin and non-Hodgkin lymphoma, EBV lymphoproliferative disease, hemophagocytic lymphohistiocytosis, and dysgammaglobulinemia. In this review, we give an update on further reported patients. We believe that current clinical data advocate early definitive treatment by hematopoietic stem cell transplantation, as transplant outcome in primary immunodeficiency disorders in general has gradually improved in recent years. Furthermore, we summarize experimental data in the murine model to provide further insight of pathophysiology in ITK deficiency.


Assuntos
Infecções por Vírus Epstein-Barr/complicações , Síndromes de Imunodeficiência/enzimologia , Transtornos Linfoproliferativos/imunologia , Proteínas Tirosina Quinases/deficiência , Adolescente , Animais , Criança , Pré-Escolar , Modelos Animais de Doenças , Infecções por Vírus Epstein-Barr/imunologia , Feminino , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 4/imunologia , Doença de Hodgkin , Humanos , Síndromes de Imunodeficiência/diagnóstico , Recém-Nascido , Linfoma não Hodgkin/complicações , Masculino , Camundongos , Camundongos Knockout , Proteínas Tirosina Quinases/imunologia , Linfócitos T/imunologia
9.
Mol Med Rep ; 18(2): 1415-1422, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29901123

RESUMO

Neutrophil­to­lymphocyte ratio (NLR) is commonly considered a useful prognostic index for many cardiovascular diseases; however, it has limited sensitivity and specificity. Factors associated with elevated NLR may aid in the prediction of prognosis with heart failure (HF) in combination with NLR. The present study sought to identify decisive factors associated with NLR in HF patients and investigate their association with elevated NLR. The gene expression profile for blood samples from 197 individuals with chronic heart failure (CHF), with corresponding hematological parameters and clinical data were obtained from the public database, GSE77343. Differentially expressed genes (DEGs) were identified, and Gene Ontology and pathway enrichment analyses were performed. The protein­protein interaction network was constructed with the Search Tool for the Retrieval of Interacting Genes along with Cytoscape. Receiver operating characteristic curves for predictive power, sensitivity and specificity were constructed. The present study identified specific associated DEGs by using Pearson linear correlation and logistic regression analysis. A mean NLR of 3.96 was determined as the cutoff value in the analysis. In total, 31 genes were initially identified as DEGs associated with elevated NLR. They were mainly enriched in neutrophil activation and neutrophil mediated immunity, in fluid shear stress and atherosclerosis, and transcriptional misregulation in cancer. Three focused DEGs, solute carrier family 22 member 4 (SLC22A4), interleukin­1 receptor 2 (IL1R2) and vanin 3 (VNN3), were finally revealed to be independently associated with elevated NLR in CHF patients. The present study demonstrated that the three genes SLC22A4, IL1R2 and VNN3 may be independently associated with elevated NLR in CHF patients as potential decisive factors of NLR.


Assuntos
Amidoidrolases/genética , Moléculas de Adesão Celular/genética , Insuficiência Cardíaca/genética , Linfócitos/metabolismo , Neutrófilos/metabolismo , Proteínas de Transporte de Cátions Orgânicos/genética , Receptores Tipo II de Interleucina-1/genética , Idoso , Amidoidrolases/imunologia , Anexina A3/genética , Anexina A3/imunologia , Biomarcadores , Moléculas de Adesão Celular/imunologia , Doença Crônica , Bases de Dados Genéticas , Feminino , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/imunologia , Regulação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Insuficiência Cardíaca/diagnóstico , Insuficiência Cardíaca/imunologia , Insuficiência Cardíaca/patologia , Humanos , Contagem de Leucócitos , Linfócitos/imunologia , Linfócitos/patologia , Masculino , Pessoa de Meia-Idade , Anotação de Sequência Molecular , Neutrófilos/imunologia , Neutrófilos/patologia , Proteínas de Transporte de Cátions Orgânicos/imunologia , Mapeamento de Interação de Proteínas , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Receptores Tipo I de Interleucina-1/genética , Receptores Tipo I de Interleucina-1/imunologia , Receptores Tipo II de Interleucina-1/imunologia
10.
Gene Ther ; 25(5): 359-375, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29907877

RESUMO

Previously, we reported that electroporation-mediated (EP) delivery of the FER gene improved survival in a combined trauma-pneumonia model. The mechanism of this protective effect is unknown. In this paper, we performed a pneumonia model in C57/BL6 mice with 500 CFU of Klebsiella pneumoniae. After inoculation, a plasmid encoding human FER was delivered by EP into the lung (PNA/pFER-EP). Survival of FER-treated vs. controls (PNA; PNA/EP-pcDNA) was recorded. In parallel cohorts, bronchial alveolar lavage (BAL) and lung were harvested at 24 and 72 h with markers of infection measured. FER-EP-treated animals reduced bacterial counts and had better 5-day survival compared to controls (80 vs. 20 vs. 25%; p < 0.05). Pre-treatment resulted in 100% survival. With FER, inflammatory monocytes were quickly recruited into BAL. These cells had increased surface expression for Toll-receptor 2 and 4, and increased phagocytic and myeloperoxidase activity at 24 h. Samples from FER electroporated animals had increased phosphorylation of STAT transcription factors, varied gene expression of IL1ß, TNFα, Nrf2, Nlrp3, Cxcl2, HSP90 and increased cytokine production of TNF-α, CCL-2, KC, IFN-γ, and IL-1RA. In a follow-up experiment, using Methicillin-resistant Staphylococcus aureus (MRSA) similar bacterial reduction effects were obtained with FER gene delivery. We conclude that FER overexpression improves survival through STAT activation enhancing innate immunity and accelerating bacterial clearance in the lung. This constitutes a novel mechanism of inflammatory regulation with therapeutic potential in the setting of hospital-acquired pneumonia.


Assuntos
Eletroporação/métodos , Pneumonia Bacteriana/terapia , Proteínas Tirosina Quinases/genética , Animais , Carga Bacteriana , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Terapia Genética/métodos , Humanos , Imunidade Inata/genética , Klebsiella pneumoniae/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia Bacteriana/imunologia , Pneumonia Bacteriana/microbiologia , Proteínas Tirosina Quinases/administração & dosagem , Proteínas Tirosina Quinases/biossíntese , Proteínas Tirosina Quinases/imunologia , Fator de Necrose Tumoral alfa/metabolismo
11.
J Neuroimmune Pharmacol ; 13(2): 265-276, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29550892

RESUMO

Multiple sclerosis is a neuroinflammatory degenerative disease, caused by activated immune cells infiltrating the CNS. The disease etiology involves both genetic and environmental factors. The mouse genetic locus, Eae27, linked to disease development in the experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis, was studied in order to identify contributing disease susceptibility factors and potential drug targets for multiple sclerosis. Studies of an Eae27 congenic mouse strain, revealed that genetic variation within Eae27 influences EAE development. The Abl2 gene, encoding the non-receptor tyrosine kinase Arg, is located in the 4,1 megabase pair long Eae27 region. The Arg protein plays an important role in cellular regulation and is, in addition, involved in signaling through the B- and T-cell receptors, important for the autoimmune response. The presence of a single nucleotide polymorphism causing an amino acid change in a near actin-interacting domain of Arg, in addition to altered lymphocyte activation in the congenic mice upon immunization with myelin antigen, makes Abl2/Arg a candidate gene for EAE. Here we demonstrate that the non-synonymous SNP does not change Arg's binding affinity for F-actin but suggest a role for Abl kinases in CNS inflammation pathogenesis by showing that pharmacological inhibition of Abl kinases ameliorates EAE, but not experimental arthritis.


Assuntos
Encefalomielite Autoimune Experimental/genética , Proteínas Tirosina Quinases/genética , Animais , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Camundongos , Camundongos Mutantes , Polimorfismo de Nucleotídeo Único , Proteínas Tirosina Quinases/imunologia , Proteínas Tirosina Quinases/metabolismo
12.
Fish Shellfish Immunol ; 77: 91-99, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29567142

RESUMO

Bruton's tyrosine kinase (BTK) is a Tec-family tyrosine kinase and plays a crucial role in B cell antigen receptor (BCR) signal pathway. Mutations in humans and mice BTK gene results in X-linked agammaglobulinemia (XLA) and X-linked immunodeficiency (XLD), respectively. To study the function of BTK in teleost, we cloned a BTK gene from orange-spotted grouper. Homology analysis showed that the grouper BTK (EcBTK) had a high amino acid identity with other vertebrates (63%-92%) and shared the highest amino acid identity with ballan wrasse Labrus bergylta BTK. EcBTK comprises a Bruton's tyrosine kinase pleckstrin homology (PH) domain, a Tec homology (TH) domain, a Src homology 3 (SH3) domain, a Src homology 2 (SH2) domain and a Protein Kinases, catalytic (PKc) domain. Tissue distribution analysis showed that EcBTK was mainly expressed in immune organs. EcBTK was uniform distributed throughout the cytoplasm of transfected HEK293T cells and overexpression of EcBTK slightly down-regulates NF-κB activity. Ibrutinib treatment can reduce the phosphorylation level of grouper's BTK. In groupers infected with Cryptocaryon irritans, up-regulation of EcBTK were not seen in the early stage of infected skin and gill until days 14-21. The phosphorylation level of grouper BTK was significantly increased in infected skin and gill.


Assuntos
Bass/genética , Bass/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Tirosina Quinase da Agamaglobulinemia , Sequência de Aminoácidos , Animais , Cilióforos/fisiologia , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/veterinária , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Proteínas Tirosina Quinases/química , Alinhamento de Sequência/veterinária
13.
Intern Med ; 57(2): 265-268, 2018 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-29093409

RESUMO

We herein report the case of myasthenic crisis occurring in a 51-year-old man. He had experienced ptosis, increased body weight with edema, and fatigue with dyspnea. He presented at our emergency department with disturbed consciousness. He was originally diagnosed with myxedema coma, and he required artificial respiration. Because his weakness persisted and he was positive for anti-acetylcholine receptor antibodies and anti-muscle-specific tyrosine kinase antibodies, we diagnosed myasthenic crisis after various examinations. His clinical response to treatment was good and he was discharged in an ambulatory status 3 months after admission. This case demonstrates that myasthenic crisis may occur in association with myxedema.


Assuntos
Miastenia Gravis/complicações , Mixedema/complicações , Proteínas Tirosina Quinases/imunologia , Receptores Colinérgicos/imunologia , Autoanticorpos/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Miastenia Gravis/terapia , Mixedema/terapia , Respiração Artificial/efeitos adversos
14.
J Allergy Clin Immunol ; 141(5): 1774-1785.e7, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-28734845

RESUMO

BACKGROUND: Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a cell-surface protein expressed selectively on human eosinophils, mast cells, and basophils, making it an ideal target for the treatment of diseases involving these cell types. However, the effective delivery of therapeutic agents to these cells requires an understanding of the dynamics of Siglec-8 surface expression. OBJECTIVES: We sought to determine whether Siglec-8 is endocytosed in human eosinophils and malignant mast cells, identify mechanisms underlying its endocytosis, and demonstrate whether a toxin can be targeted to Siglec-8-bearing cells to kill these cells. METHODS: Siglec-8 surface dynamics were examined by flow cytometry using peripheral blood eosinophils, mast cell lines, and Siglec-8-transduced cells in the presence of inhibitors targeting components of endocytic pathways. Siglec-8 intracellular trafficking was followed by confocal microscopy. The ribosome-inhibiting protein saporin was conjugated to a Siglec-8-specific antibody to examine the targeting of an agent to these cells through Siglec-8 endocytosis. RESULTS: Siglec-8 endocytosis required actin rearrangement, tyrosine kinase and protein kinase C activities, and both clathrin and lipid rafts. Internalized Siglec-8 localized to the lysosomal compartment. Maximal endocytosis in Siglec-8-transduced HEK293T cells required an intact immunoreceptor tyrosine-based inhibitory motif. Siglec-8 was also shuttled to the surface via a distinct pathway. Sialidase treatment of eosinophils revealed that Siglec-8 is partially masked by sialylated cis ligands. Targeting saporin to Siglec-8 consistently caused extensive cell death in eosinophils and the human mast cell leukemia cell line HMC-1.2. CONCLUSIONS: Therapeutic payloads can be targeted selectively to eosinophils and malignant mast cells by exploiting this Siglec-8 endocytic pathway.


Assuntos
Antígenos CD/imunologia , Antígenos de Diferenciação de Linfócitos B/imunologia , Eosinófilos/imunologia , Lectinas/imunologia , Mastócitos/imunologia , Actinas/imunologia , Basófilos/imunologia , Morte Celular/imunologia , Linhagem Celular , Linhagem Celular Tumoral , Clatrina/imunologia , Endocitose/imunologia , Citometria de Fluxo/métodos , Células HEK293 , Humanos , Ligantes , Microdomínios da Membrana/imunologia , Proteína Quinase C/imunologia , Proteínas Tirosina Quinases/imunologia
15.
Proc Natl Acad Sci U S A ; 114(47): 12542-12547, 2017 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-29109251

RESUMO

The metabolic programs of functionally distinct T cell subsets are tailored to their immunologic activities. While quiescent T cells use oxidative phosphorylation (OXPHOS) for energy production, and effector T cells (Teffs) rely on glycolysis for proliferation, the distinct metabolic features of regulatory T cells (Tregs) are less well established. Here we show that the metabolic sensor LKB1 is critical to maintain cellular metabolism and energy homeostasis in Tregs. Treg-specific deletion of Lkb1 in mice causes loss of Treg number and function, leading to a fatal, early-onset autoimmune disorder. Tregs lacking Lkb1 have defective mitochondria, compromised OXPHOS, depleted cellular ATP, and altered cellular metabolism pathways that compromise their survival and function. Furthermore, we demonstrate that the function of LKB1 in Tregs is largely independent of the AMP-activated protein kinase, but is mediated by the MAP/microtubule affinity-regulating kinases and salt-inducible kinases. Our results define a metabolic checkpoint in Tregs that couples metabolic regulation to immune homeostasis and tolerance.


Assuntos
Doenças Autoimunes/imunologia , Metabolismo Energético/imunologia , Homeostase/imunologia , Tolerância Imunológica , Proteínas Serina-Treonina Quinases/imunologia , Linfócitos T Reguladores/imunologia , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/imunologia , Animais , Doenças Autoimunes/genética , Doenças Autoimunes/metabolismo , Doenças Autoimunes/patologia , Contagem de Linfócito CD4 , Proliferação de Células , Sobrevivência Celular , Metabolismo Energético/genética , Regulação da Expressão Gênica/imunologia , Linfonodos/imunologia , Linfonodos/metabolismo , Linfonodos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitocôndrias/imunologia , Mitocôndrias/patologia , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Transdução de Sinais , Baço/imunologia , Baço/metabolismo , Baço/patologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Linfócitos T Citotóxicos/patologia , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologia
16.
Curr Opin Oncol ; 29(5): 395-399, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28787285

RESUMO

PURPOSE OF REVIEW: This review aims to highlight the novel therapeutic agents in the management of brain metastases which are in various stages of clinical development. We review the results from recent clinical trials, publications and presentations at recent national and international conferences. RECENT FINDINGS: Several new systemic treatment options for brain metastases are in early or advanced clinical trials. These drugs have good intracranial and extracranial activities. As lung cancer, breast cancer, and melanoma are the three most common causes of brain metastases, most agents in clinical development are focused on these tumor types. Several of these therapies are small molecule tyrosine kinase inhibitors or monoclonal antibodies against the tyrosine kinase receptors. Another exciting development in brain metastases management is the use of immunotherapy agents. The anti-CTLA-4 and\or anti-PD-1 antibodies have shown promising intracranial activity in melanoma and nonsmall cell lung cancer patients with brain metastases. SUMMARY: Contemporary clinical trials have shown encouraging intracranial activity of newer tyrosine kinase inhibitors, monoclonal antibodies against tyrosine kinase receptors and immunotherapy agents in select group of patients with brain metastases. Further studies are needed to develop therapeutic strategies, in order to improve survival in patients with brain metastases.


Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/secundário , Neoplasias Encefálicas/enzimologia , Neoplasias Encefálicas/imunologia , Antígeno CTLA-4/antagonistas & inibidores , Humanos , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/imunologia
17.
Sci Rep ; 7(1): 9639, 2017 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-28852108

RESUMO

Scabies is a disease that harms humans and other animals that is caused by the itch mite Sarcoptes scabiei burrowing into the stratum corneum of the skin. In the early stages of scabies, symptoms are often subclinical and there are no effective diagnostic methods. Herein, we cloned, expressed and characterised an S. scabiei protein tyrosine kinase (SsPTK) and evaluated its diagnostic value as a recombinant antigen in rabbit during the early stages of Sarcoptes infestation. The SsPTK protein is ~30 kDa, lacks a signal peptide, and shares high homology with a PTK from the rabbit ear mite Psoroptes ovis cuniculi. The protein was widely distributed at the front end of mites, particularly in the chewing mouthparts and legs. Indirect ELISA using recombinant SsPTK showed good diagnostic value, with 95.2% (40/42) sensitivity and 94.1% (48/51) specificity for detecting anti-PTK antibody in serum samples from naturally-infested rabbits. More importantly, PTK ELISA could diagnose infection in the early stages (infestation for 1 week) with an accuracy of 100% (24/24). SsPTK therefore shows potential as a sensitive antigen for the early diagnosis of parasitic mite infestation.


Assuntos
Anticorpos/sangue , Antígenos/imunologia , Proteínas de Artrópodes/imunologia , Proteínas Tirosina Quinases/imunologia , Sarcoptes scabiei/enzimologia , Escabiose/diagnóstico , Testes Sorológicos/métodos , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática/métodos , Coelhos , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade
18.
Mol Med Rep ; 16(5): 6340-6345, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28849059

RESUMO

The present study aimed to evaluate the inhibitory effects of an irinotecan derivative, ZBH­1208, on brain tumors, and to explore the underlying molecular mechanisms. To determine the effects of ZBH­1208, a brain tumor mouse model was established by transplanting B22 cells. Subsequently, the visceral indices of immune organs and white blood cell counts were determined, and the effects of ZBH­1208 on the expression levels of cell cycle­related proteins were assessed by western blotting. The tumor inhibition rates of 20 and 40 mg/kg ZBH­1208 were 11.7 and 54.1%, respectively. Compared with the negative control group, ZBH­1208 barely affected visceral indices or white blood cell count. Furthermore, the expression levels of p53, p21, cyclin­dependent kinase 7 (CDK7), Wee1, phosphorylated (p)­cell division cycle 2 (CDC2) (Tyr15), p­CDC2 (Thr161) and cyclin B1 proteins were upregulated, whereas the expression levels of cyclin E were downregulated, and those of CDC2, CDK2 and CDC25C were barely altered. In conclusion, the present study demonstrated that ZBH­1208 suppressed the growth of B22 mouse brain tumor xenografts, but did not affect their visceral indices or white blood cell counts. It was suggested that ZBH­1208 exerted its effects by regulating the expression of p53, p21, Wee1, p­CDC2 (Tyr15) and cyclin E proteins.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Camptotecina/análogos & derivados , Regulação Neoplásica da Expressão Gênica , Imunidade Inata/efeitos dos fármacos , Animais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/patologia , Proteína Quinase CDC2/genética , Proteína Quinase CDC2/imunologia , Camptotecina/farmacologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/imunologia , Linhagem Celular Tumoral , Ciclina B1/genética , Ciclina B1/imunologia , Ciclina E/genética , Ciclina E/imunologia , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/imunologia , Quinases Ciclina-Dependentes/genética , Quinases Ciclina-Dependentes/imunologia , Relação Dose-Resposta a Droga , Esquema de Medicação , Vida Livre de Germes , Irinotecano , Masculino , Camundongos , Camundongos Endogâmicos ICR , Transplante de Neoplasias , Proteínas Nucleares/genética , Proteínas Nucleares/imunologia , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Transdução de Sinais , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/imunologia
19.
PLoS One ; 12(7): e0181782, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28742141

RESUMO

Bruton's tyrosine kinase (BTK) regulates critical signal transduction pathways involved in the pathobiology of rheumatoid arthritis (RA) and other autoimmune disorders. BMS-986142 is a potent and highly selective reversible small molecule inhibitor of BTK currently being investigated in clinical trials for the treatment of both RA and primary Sjögren's syndrome. In the present report, we detail the in vitro and in vivo pharmacology of BMS-986142 and show this agent provides potent and selective inhibition of BTK (IC50 = 0.5 nM), blocks antigen receptor-dependent signaling and functional endpoints (cytokine production, co-stimulatory molecule expression, and proliferation) in human B cells (IC50 ≤ 5 nM), inhibits Fcγ receptor-dependent cytokine production from peripheral blood mononuclear cells, and blocks RANK-L-induced osteoclastogenesis. Through the benefits of impacting these important drivers of autoimmunity, BMS-986142 demonstrated robust efficacy in murine models of rheumatoid arthritis (RA), including collagen-induced arthritis (CIA) and collagen antibody-induced arthritis (CAIA). In both models, robust efficacy was observed without continuous, complete inhibition of BTK. When a suboptimal dose of BMS-986142 was combined with other agents representing the current standard of care for RA (e.g., methotrexate, the TNFα antagonist etanercept, or the murine form of CTLA4-Ig) in the CIA model, improved efficacy compared to either agent alone was observed. The results suggest BMS-986142 represents a potential therapeutic for clinical investigation in RA, as monotherapy or co-administered with agents with complementary mechanisms of action.


Assuntos
Artrite Experimental/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Tirosina Quinases/antagonistas & inibidores , Tirosina Quinase da Agamaglobulinemia , Animais , Formação de Anticorpos/efeitos dos fármacos , Artrite Experimental/imunologia , Artrite Experimental/patologia , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Linfócitos B/patologia , Feminino , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Camundongos Endogâmicos BALB C , Osteoclastos/efeitos dos fármacos , Osteoclastos/imunologia , Osteoclastos/patologia , Inibidores de Proteínas Quinases/farmacologia , Proteínas Tirosina Quinases/imunologia , Ligante RANK/imunologia
20.
Transl Psychiatry ; 7(6): e1154, 2017 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-28632203

RESUMO

Early identification of Alzheimer's disease (AD) risk factors would aid development of interventions to delay the onset of dementia, but current biomarkers are invasive and/or costly to assess. Validated plasma biomarkers would circumvent these challenges. We previously identified the kinase DYRK1A in plasma. To validate DYRK1A as a biomarker for AD diagnosis, we assessed the levels of DYRK1A and the related markers brain-derived neurotrophic factor (BDNF) and homocysteine in two unrelated AD patient cohorts with age-matched controls. Receiver-operating characteristic curves and logistic regression analyses showed that combined assessment of DYRK1A, BDNF and homocysteine has a sensitivity of 0.952, a specificity of 0.889 and an accuracy of 0.933 in testing for AD. The blood levels of these markers provide a diagnosis assessment profile. Combined assessment of these three markers outperforms most of the previous markers and could become a useful substitute to the current panel of AD biomarkers. These results associate a decreased level of DYRK1A with AD and challenge the use of DYRK1A inhibitors in peripheral tissues as treatment. These measures will be useful for diagnosis purposes.


Assuntos
Doença de Alzheimer/sangue , Fator Neurotrófico Derivado do Encéfalo/sangue , Homocisteína/sangue , Proteínas Serina-Treonina Quinases/sangue , Proteínas Tirosina Quinases/sangue , Idoso , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/farmacologia , Biomarcadores/sangue , Feminino , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pessoa de Meia-Idade , Proteínas Serina-Treonina Quinases/imunologia , Proteínas Tirosina Quinases/imunologia , Curva ROC
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