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1.
Appl Microbiol Biotechnol ; 103(18): 7491-7504, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31332484

RESUMO

Infectious bursal disease virus (IBDV) is the cause of an economically important highly contagious disease of poultry, and vaccines are regarded as the most beneficial interventions for its prevention. In this study, plants were used to produce a recombinant chimeric IBDV antigen for the formulation of an innovative subunit vaccine. The fusion protein (PD-FcY) was designed to combine the immunodominant projection domain (PD) of the viral structural protein VP2 with the constant region of avian IgY (FcY), which was selected to enhance antigen uptake by avian immune cells. The gene construct encoding the fusion protein was transiently expressed in Nicotiana benthamiana plants and an extraction/purification protocol was set up, allowing to reduce the contamination by undesired plant compounds/proteins. Mass spectrometry analysis of the purified protein revealed that the glycosylation pattern of the FcY portion was similar to that observed in native IgY, while in vitro assays demonstrated the ability of PD-FcY to bind to the avian immunoglobulin receptor CHIR-AB1. Preliminary immunization studies proved that PD-FcY was able to induce the production of protective anti-IBDV-VP2 antibodies in chickens. In conclusion, the proposed fusion strategy holds promises for the development of innovative low-cost subunit vaccines for the prevention of avian viral diseases.


Assuntos
Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Imunoglobulinas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vacinas Virais/biossíntese , Animais , Antígenos Virais/biossíntese , Galinhas/imunologia , Imunoglobulinas/biossíntese , Vírus da Doença Infecciosa da Bursa , Doenças das Aves Domésticas/virologia , Tabaco/genética , Vacinação , Vacinas de Subunidades/biossíntese , Proteínas Estruturais Virais/biossíntese , Proteínas Estruturais Virais/imunologia
2.
Bioengineered ; 10(1): 33-42, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-30913952

RESUMO

The diagnosis of influenza A virus is essential since it can be confused with influenza A like illness and lead to inaccurate drug prescription. In this study, the M2e peptide, a strategic antigen that is conserved in all virus subtypes, was used as a diagnostic marker of influenza A. For the first time, M2e-specific IgY antibody was covalently conjugated to alkaline phosphatase (ALP) enzyme in the presence of glutaraldehyde. The antibody-enzyme bioconjugate was characterized by fluorescence and Fourier-transform infrared spectroscopy. Subsequently, the diagnostic value of this bioconjugate was evaluated by direct sandwich ELISA using nasopharyngeal swab samples positive/negative for H1N1 and H3N2, which were previously analyzed by rRT-PCR for influenza. In conclusion, the M2e-specific IgY-ALP bioconjugate demonstrated positive results for Influenza A in samples that were diagnosed as Influenza A via the RT-PCR method.


Assuntos
Fosfatase Alcalina/química , Anticorpos Antivirais/química , Antígenos Virais/imunologia , Imunoglobulinas/química , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Influenza Humana/diagnóstico , Fosfatase Alcalina/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/isolamento & purificação , Antígenos Virais/administração & dosagem , Antígenos Virais/química , Galinhas , Reagentes para Ligações Cruzadas/química , Ensaio de Imunoadsorção Enzimática/métodos , Epitopos/química , Feminino , Glutaral/química , Humanos , Imunização , Imunoconjugados/química , Imunoglobulinas/biossíntese , Imunoglobulinas/isolamento & purificação , Vírus da Influenza A Subtipo H1N1/química , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/química , Vírus da Influenza A Subtipo H3N2/imunologia , Influenza Humana/imunologia , Influenza Humana/virologia , Nasofaringe/virologia , Peptídeos/administração & dosagem , Peptídeos/química , Peptídeos/imunologia , Espectroscopia de Infravermelho com Transformada de Fourier
3.
Toxicon ; 163: 84-92, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30914282

RESUMO

Antivenom for the treatment of bothropic snakebite is a priority for public health institutions from Latin America. An alternative to the conventional antivenom production is based on the use of egg yolk antibodies - IgY-technology - by immunizing laying hens. In this study, we produced, characterized and assessed the efficacy of IgY-based antivenoms against B. alternatus venom. Immunochemical studies (reactivity, avidity and antigen recognition pattern) as well as antivenom efficacy assays were performed. After the 3rd immunization, levels of specific IgY reached a maximum that was maintained throughout the observation period, while avidity indexes of the extracts increased after the successive immunizations. Furthermore, IgY against B. alternatus recognized protein complexes of the venom with high (>40 kDa), medium (20-40 kDa) and low (<20 kDa) molecular weights. IgY antivenoms obtained after 8 immunizations neutralized 35.65 µg of B. alternatus venom per mg of antivenom, while specific activities values ranged from 0.28 to 0.42. In conclusion, we produced and characterized IgY antivenoms capable of neutralizing the lethal activity of B. alternatus venom at a preclinical level. Thus, IgY-technology may allow the production of effective and affordable antivenoms fulfilling the urgent needs of many countries where conventional manufacture is unable to provide enough availability of antivenoms.


Assuntos
Antivenenos/biossíntese , Bothrops , Venenos de Crotalídeos/imunologia , Imunoglobulinas/biossíntese , Animais , Antivenenos/imunologia , Galinhas , Venenos de Crotalídeos/química , Venenos de Crotalídeos/toxicidade , Gema de Ovo/imunologia , Feminino , Imunoglobulinas/imunologia , Camundongos , Testes de Neutralização
4.
J Enzyme Inhib Med Chem ; 34(1): 613-619, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30727775

RESUMO

This study aimed to evaluate the effects of cinnamamides on atopic dermatitis (AD) and the mechanisms underlying these effects. To this end, the actions of two cinnamamides, (E)-3-(4-hydroxyphenyl)-N-phenylethyl acrylamide (NCT) and N-trans-coumaroyltyramine (NCPA), were determined on AD by orally administering them to mice. Oral administration of the cinnamamides ameliorated the increase in epidermal and dermal thickness as well as mast cell infiltration. Cinnamamides suppressed serum immunoglobulin (Ig) levels and expression of T-helper (Th)1/Th2 cytokines. Moreover, cinnamamides suppressed interleukin (IL)-4, which plays a crucial role in preparing naïve clusters of differentiation (CD)4+ T cells, and decreased the cervical lymph node size and weight. Interestingly, in almost all cases, NCPA exhibited higher anti-AD activity compared to NCT. These results strongly indicate that NCPA may have potential as an anti-AD agent, and further mechanistic comparative studies of NCT and NCPA are required to determine the cause of differences in biological activity.


Assuntos
Linfócitos T CD4-Positivos/efeitos dos fármacos , Cinamatos/farmacologia , Dermatite Atópica/tratamento farmacológico , Interleucina-4/antagonistas & inibidores , Administração Oral , Animais , Linfócitos T CD4-Positivos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Cinamatos/administração & dosagem , Cinamatos/química , Relação Dose-Resposta a Droga , Imunoglobulinas/biossíntese , Interleucina-4/metabolismo , Linfonodos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Tamanho do Órgão/efeitos dos fármacos , Relação Estrutura-Atividade
5.
Mol Immunol ; 107: 79-83, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30665061

RESUMO

Salmonella species have been the major foodborne problems in food production systems, with Salmonella enterica serovars typhimurium (S. typhimurium) and enteritidis (S. enteritidis) being among the more common isolates. The oral administration of chicken egg yolk specific antibodies (IgYs) has been established as an efficient alternative for treatment and prevention of gastrointestinal pathogens including Salmonella. The present study was aimed to investigate the possible production of specific IgYs against Salmonella typhimurium and Salmonella enteritidis in quail egg yolks. Salmonella spp.-free female Japanese quails (Coturnix coturnix japonica) were intramuscularly immunized with formalin or heat-inactivated Salmonella immunogens (1.0 × 109 CFU/mL) emulsified with Freund adjuvants. Egg yolk IgYs were purified using ammonium sulfate precipitation method. Anti-Salmonella IgYs titer and specificity were determined using enzyme-linked immunosorbent assay (ELISA) and western blot analysis. Salmonella specific IgYs detected in the immunized quails were significantly higher than those of the control group, which confirmed the immunization procedure. Specific IgYs against S. typhimurium and S. enteritidis were identified in both groups immunized with heat or formalin-inactivated immunogens. However, formalin-inactivated immunogens induced relatively higher immune responses over the heat-inactivated ones. Quail anti-Salmonella IgYs showed a high specificity to their corresponding immunogens, with moderate cross-reactivity to other members of Enterobacteriaceae family. Quail can be regarded as a valuable and inexpensive source for producing large-scale of specific antibodies that can be used for immunodiagnostic and immunotherapeutic purposes.


Assuntos
Imunoglobulinas/biossíntese , Imunoglobulinas/isolamento & purificação , Codorniz/imunologia , Salmonella enteritidis/fisiologia , Salmonella typhimurium/fisiologia , Animais , Especificidade de Anticorpos/imunologia , Reações Cruzadas/imunologia , Gema de Ovo/metabolismo
6.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30630879

RESUMO

The reductionist approach is prevalent in biomedical science. However, increasing evidence now shows that biological systems cannot be simply considered as the sum of its parts. With experimental, technological, and computational advances, we can now do more than view parts in isolation, thus we propose that an increasing holistic view (where a protein is investigated as much as a whole as possible) is now timely. To further advocate this, we review and discuss several studies and applications involving allostery, where distant protein regions can cross-talk to influence functionality. Therefore, we believe that an increasing big picture approach holds great promise, particularly in the areas of antibody engineering and drug discovery in rational drug design.


Assuntos
Formação de Anticorpos/genética , Descoberta de Drogas/tendências , Imunoglobulinas/genética , Engenharia de Proteínas/tendências , Regulação Alostérica/genética , Humanos , Imunoglobulinas/biossíntese
7.
Br J Cancer ; 119(10): 1278-1287, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30318510

RESUMO

BACKGROUND: Cytosolic deacetylase histone deacetylase 6 (HDAC6) is involved in the autophagy degradation pathway of malformed proteins, an important survival mechanism in cancer cells. We evaluated modulation of autophagy-related proteins and cell death by the HDAC6-selective inhibitor C1A. METHODS: Autophagy substrates (light chain-3 (LC-3) and p62 proteins) and endoplasmic reticulum (ER) stress phenotype were determined. Caspase-3/7 activation and cellular proliferation assays were used to assess consequences of autophagy modulation. RESULTS: C1A potently resolved autophagy substrates induced by 3-methyladenine and chloroquine. The mechanism of autophagy inhibition by HDAC6 genetic knockout or C1A treatment was consistent with abrogation of autophagosome-lysosome fusion, and decrease of Myc protein. C1A alone or combined with the proteasome inhibitor, bortezomib, enhanced cell death in malignant cells, demonstrating the complementary roles of the proteasome and autophagy pathways for clearing malformed proteins. Myc-positive neuroblastoma, KRAS-positive colorectal cancer and multiple myeloma cells showed marked cell growth inhibition in response to HDAC6 inhibitors. Finally, growth of neuroblastoma xenografts was arrested in vivo by single agent C1A, while combination with bortezomib slowed the growth of colorectal cancer xenografts. CONCLUSIONS: C1A resolves autophagy substrates in malignant cells and induces cell death, warranting its use for in vivo pre-clinical autophagy research.


Assuntos
Autofagia/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Desacetilase 6 de Histona/antagonistas & inibidores , Inibidores de Histona Desacetilases/farmacologia , Ácidos Hidroxâmicos/farmacologia , Animais , Antineoplásicos/farmacologia , Bortezomib/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Xenoenxertos , Humanos , Imunoglobulinas/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , Ratos , Vorinostat/farmacologia , Proteínas ras/genética , Proteínas ras/metabolismo
8.
Cell Mol Biol (Noisy-le-grand) ; 64(4): 108-112, 2018 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-29631692

RESUMO

In this study, the effect of Tunceli garlic (Allium tuncelianium) on hemoglobin (Hb) level, oxidative radical production of neutrophils (Nitoblue tetrazolium assay-NBT activity) and total immunoglobulin (TI) content in carp (Cyprinus carpio) exposed to chlorpyrifos (CPF)  was investigated. The 96 hour LC50 value of CPF on C. carpio was calculated to as 0.230 mg/L. The fishes were exposed to sublethal concentration of chlorpyrifos (1/8 of LC50 value: 0.029 mg/L), and Tunceli garlic (20 and 40 g/kg diet) was simultaneously administered for 21 days. Blood samples were taken from the fishes at 7, 14 and 21 days and analysed to determine the Hb levels, the NBT activity and the TI content. There was a significant decrease in the Hb level, the NBT activity and the TI content of CPF-treated fish. However, Tunceli garlic reversed the Hb level, the NBT activity and the TI content. In conclusion, this study demonstrated that CPF had a negative effect on the immunological values of the fish. The simultaneous administration of Tunceli garlic was neutralised CPF-induced toxicity.


Assuntos
Antioxidantes/farmacologia , Clorpirifos/antagonistas & inibidores , Alho/química , Inseticidas/antagonistas & inibidores , Neutrófilos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Antioxidantes/química , Carpas/imunologia , Carpas/metabolismo , Clorpirifos/toxicidade , Hemoglobinas/metabolismo , Imunoglobulinas/biossíntese , Inseticidas/toxicidade , Dose Letal Mediana , Neutrófilos/citologia , Neutrófilos/imunologia , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Poluentes Químicos da Água/antagonistas & inibidores , Poluentes Químicos da Água/toxicidade
9.
Immunology ; 153(4): 502-512, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29044495

RESUMO

Epidemiological studies have consistently shown associations between elevated concentrations of urban particulate matter (UPM) air pollution and exacerbations of asthma and chronic obstructive pulmonary disease, which are both associated with viral respiratory infections. The effects of UPM on dendritic cell (DC) -stimulated CD4 T lymphocytes have been investigated previously, but little work has focused on CD8 T-lymphocyte responses despite their importance in anti-viral immunity. To address this, we examined the effects of UPM on DC-stimulated naive CD8 T-cell responses. Expression of the maturation/activation markers CD83, CCR7, CD40 and MHC class I on human myeloid DCs (mDCs) was characterized by flow cytometry after stimulation with UPMin vitro in the presence/absence of granulocyte-macrophage colony-stimulating factor (GM-CSF). The capacity of these mDCs to stimulate naive CD8 T-lymphocyte responses in allogeneic co-culture was then assessed by measuring T-cell cytokine secretion using cytometric bead array, and proliferation and frequency of interferon-γ (IFN-γ)-producing T lymphocytes by flow cytometry. Treatment of mDCs with UPM increased expression of CD83 and CCR7, but not MHC class I. In allogeneic co-cultures, UPM treatment of mDCs enhanced CD8 T-cell proliferation and the frequency of IFN-γ+ cells. The secretion of tumour necrosis factor-α, interleukin-13, Granzyme A and Granzyme B were also increased. GM-CSF alone, and in concert with UPM, enhanced many of these T-cell functions. The PM-induced increase in Granzyme A was confirmed in a human experimental diesel exposure study. These data demonstrate that UPM treatment of mDCs enhances priming of naive CD8 T lymphocytes and increases production of pro-inflammatory cytokines. Such UPM-induced stimulation of CD8 cells may potentiate T-lymphocyte cytotoxic responses upon concurrent airway infection, increasing bystander damage to the airways.


Assuntos
Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/efeitos dos fármacos , Material Particulado/farmacologia , Antígenos CD/biossíntese , Antígenos CD/imunologia , Proliferação de Células , Células Cultivadas , Células Dendríticas/imunologia , Voluntários Saudáveis , Humanos , Imunoglobulinas/biossíntese , Imunoglobulinas/imunologia , Glicoproteínas de Membrana/biossíntese , Glicoproteínas de Membrana/imunologia , Material Particulado/química , Receptores CCR7/biossíntese , Receptores CCR7/imunologia
10.
Cell Immunol ; 323: 41-48, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29100594

RESUMO

Intestinal immunoglobulins (Igs) protect against microbes. However, the regulation of intestinal Ig production is poorly understood. In this study, we have investigated the roles of APRIL (tumor necrosis factor superfamily member [TNFSF] 13) and BAFF (TNFSF13B) in intestinal Ig induction. Peyer's patches (PPs) are, at least in part, an inductive site for Igs, including IgA. Introducing APRIL and BAFF in vivo lowered the frequency of IgG1+ or IgG2b+ B cells in PPs. Administration of TACI-Fc upregulated the frequency of IgG1+, IgG2b+, and IgA+ B cells in PPs, suggesting that APRIL and BAFF attenuate Ig production in these regions. TACI-Fc also upregulated intestinal IgA levels and expanded germinal center B cells in PPs. These results indicate that APRIL and BAFF paradoxically downregulate homeostatic Ig production in the intestines.


Assuntos
Fator Ativador de Células B/metabolismo , Imunoglobulinas/biossíntese , Intestinos/imunologia , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo , Animais , Formação de Anticorpos , Fator Ativador de Células B/imunologia , Receptor do Fator Ativador de Células B/metabolismo , Antígeno de Maturação de Linfócitos B/metabolismo , Linfócitos B/imunologia , Regulação para Baixo , Feminino , Homeostase/imunologia , Imunoglobulina A/metabolismo , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Imunoglobulinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nódulos Linfáticos Agregados/imunologia , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/imunologia , Fator de Necrose Tumoral alfa/metabolismo
11.
Anat Histol Embryol ; 47(1): 46-50, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29152785

RESUMO

Ultrastructure of plasma cells in Harderian gland was investigated using the transmission electron microscopy. For this research, we examined the glands of 32 laying hens collected at 1, 7, 20 and 40 days and 4, 6, 8 and 12 months of the birds' ages. The research showed that the stroma of the gland contains a large number of lymphocytes and plasma cells. Most of the plasma cells are mature, but morphologically do not show productive activity. Only some individual plasma cells, situated under the secretory epithelium of primary and secondary ducts, have extremely dilated cisternae of rough endoplasmic reticulum which contain moderately dense, granular material. The morphology of these cells indicates that they are in active stage of immunoglobulin production. Also, we identified plasma cells with two types of Russell bodies. One type of these bodies was small, round or oval, while the other had irregular, angular shape. It was noted that one plasma cell never contains both type of Russell bodies at the same time. These cells were often affected by apoptosis. Among them, in deeper part of the stroma, were situated the small plasmablast cells.


Assuntos
Galinhas/anatomia & histologia , Glândula de Harder/citologia , Plasmócitos/ultraestrutura , Fatores Etários , Animais , Núcleo Celular/ultraestrutura , Retículo Endoplasmático/ultraestrutura , Feminino , Complexo de Golgi/ultraestrutura , Glândula de Harder/ultraestrutura , Imunoglobulinas/biossíntese , Microscopia Eletrônica de Transmissão/veterinária , Plasmócitos/imunologia
12.
Artif Cells Nanomed Biotechnol ; 46(1): 56-61, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28278575

RESUMO

The lipopolysaccharide (LPS) of Vibrio cholerae (V. cholerae) plays an important role in cholera disease and the induction of primary protection. In this study, we evaluate mice humoral immune response in intranasal and intraperitoneal administrated V. cholerae LPS. The results showed that the intranasal administration of LPS-chitosan nanoparticle induced the high level of antibodies compared to intraperitoneal injection of antigen without chitosan (P < .001). These results indicated that intranasal and intraperitoneal administration of LPS has been able to induce the high level of antibodies both in the sera and lavage fluid and confirmed our strategy for using intranasal administration of antigen.


Assuntos
Quitosana/química , Portadores de Fármacos/química , Imunoglobulinas/biossíntese , Lipopolissacarídeos/química , Lipopolissacarídeos/farmacologia , Nanopartículas/química , Vibrio cholerae/química , Animais , Feminino , Imunidade Humoral/efeitos dos fármacos , Imunização , Imunoglobulinas/sangue , Injeções , Camundongos , Camundongos Endogâmicos BALB C
13.
Acta Vet Scand ; 59(1): 80, 2017 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-29208016

RESUMO

BACKGROUND: Campylobacter jejuni is a major cause of foodborne disease having chickens as an important reservoir. Its control at the farm would lower the contamination of the final products and therefore also lower the risk of transmission to humans. At the farm, C. jejuni is rarely found in chickens before they reach 2 weeks of age. Past studies have shown that maternal antibodies could hamper C. jejuni gut colonization. The objective of this study was to compare protocols to use in order to produce anti-C. jejuni antibodies derived from egg yolks in the perspective to be used as feed additives for the control of chicken C. jejuni colonization. Laying hens were naturally contaminated with four well-characterized strains or injected with either outer membrane proteins or formalin-killed whole bacteria derived from these same strains. Eggs were collected and IgYs present in the yolks were extracted. The amount and the specificity of the recovered antibodies were characterized. RESULTS: It was observed that injection yielded eggs with superior concentrations of both total and anti-C. jejuni antibodies. Equivalent performances for antibodies recovered from all protocols were observed for the ability of the antibodies to agglutinate the live C. jejuni homologous strains, to hinder their motility or to lyse the bacteria. Western blot analyses showed that proteins from all strains could be recognized by all IgY extracts. All these characteristics were strain specific. The characterization assays were also made for heterologous strains and weaker results were observed when compared to the homologous strains. CONCLUSIONS: Based on these results, only an IgY quantitative based selection can be made in regards to which protocol would give the best anti-C. jejuni IgY enriched egg-yolks as all tested protocols were equivalent in terms of the recovered antibody ability to recognized the tested C. jejuni strains.


Assuntos
Infecções por Campylobacter/veterinária , Gema de Ovo , Imunoglobulinas/biossíntese , Imunoglobulinas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Animais , Proteínas de Bactérias/metabolismo , Infecções por Campylobacter/imunologia , Infecções por Campylobacter/prevenção & controle , Campylobacter jejuni/imunologia , Galinhas , Gema de Ovo/imunologia , Feminino , Doenças das Aves Domésticas/imunologia
14.
Virchows Arch ; 471(5): 659-666, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28940014

RESUMO

Peripheral T cell lymphoma, not otherwise specified (PTCL-NOS), is a heterogeneous disease with respect to clinicopathological features. Cell adhesion molecule 1 (CADM1) has been reported to be ectopically expressed in adult T cell leukaemia/lymphoma (ATLL). However, the frequency of CADM1 expression remains unknown in peripheral T cell lymphomas. In the current study, CADM1 expression was analysed in 88 PTCL-NOS patients. CADM1 was expressed in 14 of 88 (15.9%) PTCL-NOS cases, and its expression was associated with C-C chemokine receptor type 4 (CCR4) expression and nuclear atypia. CADM1-positive PTCL-NOS cases (10/74) had a significantly poorer prognosis than CADM1-negative cases (64/74) (P = 0.001). Multivariate analysis confirmed that CADM1 expression was an independent prognostic factor in PTCL-NOS. These findings suggest that CADM1 expression is a novel prognostic factor for PTCL-NOS.


Assuntos
Biomarcadores Tumorais/análise , Moléculas de Adesão Celular/biossíntese , Imunoglobulinas/biossíntese , Linfoma de Células T Periférico/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Molécula 1 de Adesão Celular , Moléculas de Adesão Celular/análise , Criança , Pré-Escolar , Feminino , Humanos , Imunoglobulinas/análise , Estimativa de Kaplan-Meier , Linfoma de Células T Periférico/metabolismo , Linfoma de Células T Periférico/mortalidade , Masculino , Pessoa de Meia-Idade , Prognóstico , Adulto Jovem
15.
J Immunol Methods ; 449: 56-61, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28697990

RESUMO

Trypanosoma cruzi is a flagellated protozoan belonging to the Trypanosomatidae family, the etiologic agent of Chagas disease. Currently, there is neither a licensed vaccine nor effective treatment, characterizing an unmet clinical need. The IgY refers to the egg yolk immunoglobulin (Y=yolk) and its production and use are subjects of many studies due to the diversity of its diagnostic and therapeutic applications. Several researchers have shown that the use of specific IgY may prevent and/or control infectious and parasitic diseases. Based on these evidences, the aim of this study was to immunize chickens with trypomastigotes of T. cruzi in order to produce highly effective and pure antibodies (IgY), as well as extract, characterize, quantify, and verify cytotoxic effects of IgY anti-T. cruzi. After the induction of IgY production by chickens, the eggs were collected and the IgY was extracted by method of precipitation of polyethylene glycol 6000. The IgY anti-T. cruzi characterization was performed using polyacrylamide gel electrophoresis (SDS-PAGE), western-blot and enzyme-linked immunosorbent assay (ELISA). Moreover, the cytotoxic or proliferative effects of IgY anti-T. cruzi was verified by MTT assay. The concentration of IgY in yolk was 8.41±1.47mg/mL. The characterization of IgY reveled bands of stained peptides with molecular weight between 75 and 50kDa and 37 and 25kDa. In the ELISA test was observed that there was antigen-antibody reaction throughout the sample period. The concentrations of 1, 5 and 10mg/mL of IgY anti-T. cruzi presented no cytotoxic of proliferative effects in mononuclear and VERO cells in vitro. The results indicated that T. cruzi is able to generate a high production of specific immunoglobulins in chickens, it did not cause damage to the cell membrane and no proliferative effect.


Assuntos
Anticorpos Antiprotozoários/imunologia , Anticorpos Antiprotozoários/isolamento & purificação , Galinhas/imunologia , Imunoglobulinas/imunologia , Imunoglobulinas/isolamento & purificação , Trypanosoma cruzi/imunologia , Animais , Anticorpos Antiprotozoários/biossíntese , Western Blotting , Testes Imunológicos de Citotoxicidade , Gema de Ovo/química , Gema de Ovo/imunologia , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Imunização , Imunoglobulinas/biossíntese , Células Vero
16.
Eur J Neurol ; 24(9): 1188-1190, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28677890

RESUMO

BACKGROUND AND PURPOSE: To compare the frequency of intrathecal immunoglobulin (Ig) synthesis in patients with symptomatic epilepsy and epilepsy of unknown etiology ('cryptogenic'). METHODS: Patients with epileptic (n = 301) and non-epileptic (n = 10) seizures were retrospectively screened for autochthonous intrathecal Ig synthesis and oligoclonal bands (OCBs) in the cerebrospinal fluid. RESULTS: Intrathecal IgG/OCBs were detected in 8% of patients with epilepsies of unknown etiology, 5% of patients with first seizures of unknown cause and 0-4% of patients with epilepsy due to brain tumors, cerebrovascular disease or other etiologies. Intrathecal IgG/OCBs were not seen in patients with psychogenic seizures. Identical OCBs in serum and cerebrospinal fluid were more common in all patient groups (10-40% depending on underlying etiology). CONCLUSIONS: Intrathecal IgG synthesis/OCBs were observed slightly more frequently in patients with 'cryptogenic' epilepsy and with first seizures of unknown etiology than in other patient groups. However, this remained an infrequent finding and thus we could not confirm humoral immunity as a leading disease mechanism in patients with epilepsy in general or with unknown etiology in particular.


Assuntos
Epilepsia/líquido cefalorraquidiano , Imunoglobulinas/líquido cefalorraquidiano , Medula Espinal/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Epilepsia/etiologia , Epilepsia/imunologia , Feminino , Humanos , Imunoglobulina A/biossíntese , Imunoglobulina A/líquido cefalorraquidiano , Imunoglobulina G/biossíntese , Imunoglobulina G/líquido cefalorraquidiano , Imunoglobulina M/biossíntese , Imunoglobulina M/líquido cefalorraquidiano , Imunoglobulinas/biossíntese , Masculino , Pessoa de Meia-Idade , Bandas Oligoclonais/líquido cefalorraquidiano , Bandas Oligoclonais/imunologia , Estudos Retrospectivos , Convulsões/líquido cefalorraquidiano , Convulsões/imunologia , Convulsões/metabolismo , Adulto Jovem
17.
J Virol ; 91(18)2017 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-28659471

RESUMO

Porcine reproductive and respiratory syndrome, caused by porcine reproductive and respiratory syndrome virus (PRRSV), is a panzootic disease that is one of the most economically costly diseases to the swine industry. A key aspect of PRRSV virulence is that the virus suppresses the innate immune response and induces persistent infection, although the underlying mechanisms are not well understood. The dendritic cell (DC) marker CD83 belongs to the immunoglobulin superfamily and is associated with DC activation and immunosuppression of T cell proliferation when expressed as soluble CD83 (sCD83). In this study, we show that PRRSV infection strongly stimulates CD83 expression in porcine monocyte-derived DCs (MoDCs) and that the nucleocapsid (N) protein and nonstructural protein 10 (nsp10) of PRRSV enhance CD83 promoter activity via the NF-κB and Sp1 signaling pathways. R43A and K44A amino acid substitution mutants of the N protein suppress the N protein-mediated increase of CD83 promoter activity. Similarly, P192-5A and G214-3A mutants of nsp10 (with 5 and 3 alanine substitutions beginning at residues P192 and G214, respectively) abolish the nsp10-mediated induction of the CD83 promoter. Using reverse genetics, four mutant viruses (rR43A, rK44A, rP192-5A, and rG214-3A) and four revertants [rR43A(R), rK44A(R), rP192-5A(R), and rG214-3A(R)] were generated. Decreased induction of CD83 in MoDCs was observed after infection by mutants rR43A, rK44A, rP192-5A, and rG214-3A, in contrast to the results obtained using rR43A(R), rK44A(R), rP192-5A(R), and rG214-3A(R). These findings suggest that PRRSV N and nsp10 play important roles in modulating CD83 signaling and shed light on the mechanism by which PRRSV modulates host immunity.IMPORTANCE Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically costly pathogens affecting the swine industry. It is unclear how PRRSV inhibits the host's immune response and induces persistent infection. The dendritic cell (DC) marker CD83 belongs to the immunoglobulin superfamily and has previously been associated with DC activation and immunosuppression of T cell proliferation and differentiation when expressed as soluble CD83 (sCD83). In this study, we found that PRRSV infection induces sCD83 expression in porcine MoDCs via the NF-κB and Sp1 signaling pathways. The viral nucleocapsid protein, nonstructural protein 1 (nsp1), and nsp10 were shown to enhance CD83 promoter activity. Amino acids R43 and K44 of the N protein, as well as residues 192 to 196 (P192-5) and 214 to 216 (G214-3) of nsp10, play important roles in CD83 promoter activation. These findings provide new insights into the molecular mechanism of immune suppression by PRRSV.


Assuntos
Antígenos CD/biossíntese , Imunoglobulinas/biossíntese , Glicoproteínas de Membrana/biossíntese , NF-kappa B/metabolismo , Proteínas do Nucleocapsídeo/metabolismo , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Proteínas Quinases/metabolismo , Transdução de Sinais , Proteínas não Estruturais Virais/metabolismo , Animais , Análise Mutacional de DNA , Células Dendríticas/imunologia , Interações Hospedeiro-Patógeno , Tolerância Imunológica , Proteínas do Nucleocapsídeo/genética , Suínos , Linfócitos T/imunologia , Regulação para Cima , Proteínas não Estruturais Virais/genética
18.
Tumour Biol ; 39(6): 1010428317704365, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28651491

RESUMO

Epithelial carcinomas occasionally have sarcomatous components that consist primarily of spindle and cuboidal cells, which often resemble osteoblasts. Sarcomatoid carcinomas consist of similar cells. Recent studies have characterized these phenomena as a manifestation of epithelial-mesenchymal transition in carcinoma cells, but the mesenchymal phenotypes that manifest in sarcomatous cells of epithelial carcinomas are not well understood. Here, we examined the expression profiles of four osteoblastic differentiation biomarkers in the sarcomatous components of multiple carcinoma types, including five renal clear cell, four breast invasive ductal, two esophageal, one maxillary squamous cell, three larynx, three lung, one liver, and one skin sarcomatoid carcinoma. Expression was analyzed by immunohistochemistry using antibodies against cell adhesion molecule 1, a member of the IgCAM superfamily, osterix transcription factor (Osterix), cluster of differentiation 151, a transmembrane 4 superfamily member, and alkaline phosphatase. Immunostaining intensity was rated in scale 0 (negative), 0.5 (weak), and 1 (strong) for each marker, and the four scale values were summed to calculate osteoblastic scores. In all, 10 cases had a osteoblastic score ≥3, and all of these 10 cases were cell adhesion molecule 1- and Osterix-positive. Eight and five of the nine samples with a osteoblastic score <3 were negative for cell adhesion molecule 1 ( p < 0.0001) and Osterix ( p = 0.006), respectively. The other markers showed no statistical significance. These results indicate that osteoblastic differentiation can occur in carcinoma cells and that cell adhesion molecule 1 could be a useful marker for identifying this phenomenon in carcinoma tissues.


Assuntos
Fosfatase Alcalina/biossíntese , Carcinoma/genética , Moléculas de Adesão Celular/biossíntese , Imunoglobulinas/biossíntese , Sarcoma/genética , Tetraspanina 24/biossíntese , Fatores de Transcrição/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Fosfatase Alcalina/genética , Biomarcadores Tumorais/biossíntese , Carcinoma/patologia , Molécula 1 de Adesão Celular , Moléculas de Adesão Celular/genética , Diferenciação Celular/genética , Transição Epitelial-Mesenquimal , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imunoglobulinas/genética , Masculino , Pessoa de Meia-Idade , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Epiteliais e Glandulares/patologia , Osteoblastos/patologia , Sarcoma/patologia , Fator de Transcrição Sp7 , Tetraspanina 24/genética , Fatores de Transcrição/genética
19.
PLoS One ; 12(4): e0173334, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28403146

RESUMO

Previous studies on a limited number of birds suggested that the IgD-encoding gene was absent in birds. However, one of our recent studies showed that the gene was definitely expressed in the ostrich and emu. Interestingly, we also identified subclass diversification of IgM and IgY in these two birds. To better understand immunoglobulin genes in birds, in this study, we analyzed the immunoglobulin heavy chain genes in the zebra finch (Taeniopygia guttata) and Gentoo penguin (Pygoscelis papua), belonging respectively to the order Passeriformes, the most successful bird order in terms of species diversity and numbers, and Sphenisciformes, a relatively primitive avian order. Similar to the results obtained in chickens and ducks, only three genes encoding immunoglobulin heavy chain isotypes, IgM, IgA and IgY, were identified in both species. Besides, we detected a transcript encoding a short membrane-bound IgA lacking the last two CH exons in the Gentoo penguin. We did not find any evidence supporting the presence of IgD gene or subclass diversification of IgM/IgY in penguin or zebra finch. The obtained data in our study provide more insights into the immunoglobulin heavy chain genes in birds and may help to better understand the evolution of immunoglobulin genes in tetrapods.


Assuntos
Proteínas Aviárias/genética , Tentilhões/genética , Imunoglobulina A/genética , Imunoglobulina M/genética , Imunoglobulinas/genética , Spheniscidae/genética , Sequência de Aminoácidos , Animais , Proteínas Aviárias/biossíntese , Sequência Conservada , Evolução Molecular , Expressão Gênica , Genes de Imunoglobulinas , Imunoglobulina A/biossíntese , Imunoglobulina M/biossíntese , Imunoglobulinas/biossíntese , Filogenia , Transcriptoma , Recombinação V(D)J
20.
PLoS One ; 12(4): e0175632, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28414795

RESUMO

Trimethylation of histone H3 lysine 4 and lysine 27 (H3K4me3 and H3K27me3) at gene promoter regions critically regulates gene expression. Key developmental genes tend to exhibit changes in histone modification patterns from the H3K4me3/H3K27me3 bivalent pattern to the H3K4me3 monovalent pattern. Using comprehensive chromatin immunoprecipitation followed by sequencing in bone marrow-derived macrophages (BMMs) and mature osteoclasts, we found that cell surface adhesion molecule 1 (Cadm1) is a direct target of nuclear factor of activated T cells 1 (NFATc1) and exhibits a bivalent histone pattern in BMMs and a monovalent pattern in osteoclasts. Cadm1 expression was upregulated in BMMs by receptor activator of nuclear factor kappa B ligand (RANKL), and blocked by a calcineurin/NFATc1 inhibitor, FK506. Cadm1-deficient mice exhibited significantly reduced bone mass compared with wild-type mice, which was due to the increased osteoclast differentiation, survival and bone-resorbing activity in Cadm1-deficient osteoclasts. These results suggest that Cadm1 is a direct target of NFATc1, which is induced by RANKL through epigenetic modification, and regulates osteoclastic bone resorption in a negative feedback manner.


Assuntos
Reabsorção Óssea/metabolismo , Moléculas de Adesão Celular/biossíntese , Imunoglobulinas/biossíntese , Fatores de Transcrição NFATC/metabolismo , Animais , Reabsorção Óssea/genética , Reabsorção Óssea/patologia , Molécula 1 de Adesão Celular , Moléculas de Adesão Celular/deficiência , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Diferenciação Celular , Epigênese Genética , Retroalimentação Fisiológica , Expressão Gênica , Histonas/genética , Histonas/metabolismo , Imunoglobulinas/deficiência , Imunoglobulinas/genética , Macrófagos/citologia , Macrófagos/metabolismo , Masculino , Metilação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Osteoclastos/citologia , Osteoclastos/metabolismo , Regiões Promotoras Genéticas , Ligante RANK/metabolismo
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