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1.
J Anim Sci ; 97(10): 4362-4369, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31504608

RESUMO

This study utilized 204 Angus-based beef steers (249 ± 23 kg SD) from a single ranch with initial serum α-tocopherol concentrations of 3.9 ± 1.0 mg/L to determine the effect of varying doses of vitamin E (VE) on feedlot performance, antibody response to vaccination, and antioxidant defense. Seven days after arrival, steers were blocked by body weight and weaning protocol (preweaned, unweaned heavy, and unweaned light) and randomly assigned to pens within blocks (12 pens per block). Preweaned steers had been weaned for approximately 35 d prior to arrival, and unweaned steers were weaned when leaving the origin ranch. Pens within block were randomly assigned to supplemental VE (ROVIMIX E-50 Adsorbate, DSM Nutritional Products, Heerlen, The Netherlands) treatments (n = 9 pens per treatment): no supplemental VE (CON), 25 IU/kg dry matter (DM; LOW), 500 IU per steer daily (MED), or 1,000 IU per steer daily (HIGH). Back-calculated supplemental VE intake was 0, 151 (24.8 IU/kg DM), 484, and 995 IU/d for CON, LOW, MED, and HIGH, respectively. On day 6, all steers received a booster vaccine against bovine viral diarrhea virus (BVDV; Bovi-Shield Gold, One Shot, Zoetis, Parsippany, NJ). Steers were weighed on day -1, 0, 14, 26, and 27. One steer per pen representative of the average body weight of the pen was chosen as a sampling animal for blood (day -1, 6, 14, 26, and 28) and liver (day -3 and 24). Data were analyzed as a randomized complete block design using Proc Mixed of SAS with pen as the experimental unit and the fixed effects of treatment and block. Linear, quadratic, and cubic contrast statements were constructed using Proc IML; morbidity data were analyzed using Proc Glimmix. Day 24 liver and day 26 serum α-tocopherol concentrations were linearly increased by supplemental VE (P < 0.01). Supplemental VE did not affect DM intake, average daily gain, or gain:feed from day 0 to 27 (P ≥ 0.37), or the percentage of steers treated for respiratory disease (P ≥ 0.44). Day 24 liver glutathione concentrations decreased linearly due to supplemental VE (P ≤ 0.02). Total- and Mn-superoxide dismutase activities were quadratically affected by supplemental VE (P ≤ 0.07), with MED steers exhibiting the greatest activity. Over time, BVDV type 1 and 2 antibody titers numerically decreased, whereas the decrease in BVDV type 1 titers was lesser for HIGH steers (linear P = 0.04). Increasing doses of VE improved VE status but did not affect overall receiving period performance in steers with minimal to adequate VE status upon arrival.


Assuntos
Antioxidantes/farmacologia , Bovinos/fisiologia , Suplementos Nutricionais/análise , Vacinação/veterinária , Vitamina E/farmacologia , Ração Animal/análise , Animais , Formação de Anticorpos/efeitos dos fármacos , Peso Corporal , Bovinos/crescimento & desenvolvimento , Bovinos/imunologia , Dieta/veterinária , Fígado/efeitos dos fármacos , Masculino , Distribuição Aleatória , Desmame
2.
Nat Med ; 25(9): 1402-1407, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31501610

RESUMO

Natalizumab (NZM), a humanized monoclonal IgG4 antibody to α4 integrins, is used to treat patients with relapsing-remitting multiple sclerosis (MS)1,2, but in about 6% of the cases persistent neutralizing anti-drug antibodies (ADAs) are induced leading to therapy discontinuation3,4. To understand the basis of the ADA response and the mechanism of ADA-mediated neutralization, we performed an in-depth analysis of the B and T cell responses in two patients. By characterizing a large panel of NZM-specific monoclonal antibodies, we found that, in both patients, the response was polyclonal and targeted different epitopes of the NZM idiotype. The neutralizing activity was acquired through somatic mutations and correlated with a slow dissociation rate, a finding that was supported by structural data. Interestingly, in both patients, the analysis of the CD4+ T cell response, combined with mass spectrometry-based peptidomics, revealed a single immunodominant T cell epitope spanning the FR2-CDR2 region of the NZM light chain. Moreover, a CDR2-modified version of NZM was not recognized by T cells, while retaining binding to α4 integrins. Collectively, our integrated analysis identifies the basis of T-B collaboration that leads to ADA-mediated therapeutic resistance and delineates an approach to design novel deimmunized antibodies for autoimmune disease and cancer treatment.


Assuntos
Anticorpos Neutralizantes/administração & dosagem , Epitopos de Linfócito T/imunologia , Esclerose Múltipla/tratamento farmacológico , Natalizumab/administração & dosagem , Anticorpos Monoclonais Humanizados/administração & dosagem , Anticorpos Neutralizantes/química , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Linfócitos B/efeitos dos fármacos , Humanos , Imunoglobulina G/química , Imunoglobulina G/imunologia , Integrina alfa4/antagonistas & inibidores , Integrina alfa4/imunologia , Esclerose Múltipla/imunologia , Esclerose Múltipla/patologia , Conformação Proteica/efeitos dos fármacos , Linfócitos T/química , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia
3.
Food Chem Toxicol ; 132: 110696, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31344370

RESUMO

Ochratoxin A (OA), the secondary metabolite of certain Aspergillus and Penicillium species, is one of the potent biological immune-suppressor. The present study was designed to explore the in-vivo efficacy of Trichosporon mycotoxinivorans (TR); yeast strain isolated from the hindgut of the termite Mastotermes darwiniensis, against the immunotoxicity of OA in broiler birds. For this purpose, broiler chicks were offered diet added with TR (0.5, 1.0 or 2.0 g/kg feed) and/or OA (0.15, 0.3 or 1.0 mg/kg feed) for 42 days. Dietary OA at all levels, resulted in significant reduction (p ≤ 0.05) in the immune response of broiler birds as recorded by vacuolation and darkly stained pyknotic nuclei in bursa of Fabricius and thymus, humoral immune responses to sheep red blood cells (SRBC), in-vivo lymphoproliferative response to Phytohemagglutinin-P (PHA-P) and mononuclear phagocytic system function assay. Addition of TR in broiler diet significantly reduced (p ≤ 0.05) the immunotoxicity of OA at 0.15 and 0.30 mg/kg; however, against higher dietary level of OA (1.0 mg/kg), a partial protection was observed. Feeding TR alone had no immunomodulatory effect at any of tested level. Dietary addition of TR is proposed as an approach to combat the OA mediated immunological damages in broiler birds.


Assuntos
Ração Animal , Galinhas/imunologia , Ocratoxinas/toxicidade , Trichosporon , Animais , Formação de Anticorpos/efeitos dos fármacos , Basófilos/efeitos dos fármacos , Basófilos/imunologia , Relação Dose-Resposta a Droga , Ocratoxinas/administração & dosagem , Fito-Hemaglutininas/farmacologia , Ovinos
4.
Expert Opin Ther Pat ; 29(5): 339-351, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31064237

RESUMO

INTRODUCTION: Boron-containing compounds induce effects on immune responses. Such effects are interesting to the biomedical field for the development of therapeutic tools to modulate the immune system. AREAS COVERED: The scope of BCC use to modify immune responses is expanding, mainly with regard to inflammatory diseases. The information was organized to demonstrate the breadth of reported effects. BCCs act as modulators of innate and adaptive immunity, with the former including regulation of cluster differentiation and cytokine production. In addition, BCCs exert effects on inflammation induced by infectious and noninfectious agents, and there are also reports regarding their effects on mechanisms involving hypersensitivity and transplants. Finally, the authors discuss the beneficial effects of BCCs on pathologies involving various targets and mechanisms. EXPERT OPINION: Some BCCs are currently used as drugs in humans. The mechanisms by which these BCCs modulate immune responses, as well as the required structure-activity relationship for each observed mechanism of action, should be clarified. The former will allow for the development of improved immunomodulatory drugs with extensive applications in medicine. Patenting trends involve claims concerning the synthesis and actions of identified molecules with a defined profile regarding cytokines, cell differentiation, proliferation, and antibody production.


Assuntos
Compostos de Boro/farmacologia , Fatores Imunológicos/farmacologia , Inflamação/tratamento farmacológico , Imunidade Adaptativa/efeitos dos fármacos , Animais , Formação de Anticorpos/efeitos dos fármacos , Compostos de Boro/química , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocinas/imunologia , Humanos , Imunidade Inata/efeitos dos fármacos , Fatores Imunológicos/química , Inflamação/imunologia , Patentes como Assunto , Relação Estrutura-Atividade
5.
Biosci Biotechnol Biochem ; 83(10): 1791-1799, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31112075

RESUMO

To produce the antiserum against a small peptide, the target peptide-keyhole limpet hemocyanine (KLH) conjugate is generally used as an antigen, although the disulfide-rich peptide-KLH conjugate is still difficult to prepare. In our previous study, we have developed a preparation method of the disulfide-rich peptide-KLH conjugate, and this method was applied to produce the antiserum against a relaxin-like peptide. However, this method is limited to the synthetic peptide antigen, and is not applicable to a native or a recombinant peptide. In this study, to expand the applicability of this method to wide variety of peptides, we newly designed a novel thiol probe enabling the conjugation between various peptides and KLH, and applied it to produce the antiserum against relaxin-like peptide of a starfish Asterias amurensis. The antiserum obtained here showed high antibody-titer and good specificity, strongly suggesting that the method developed in this study is applicable to various peptides.


Assuntos
Formação de Anticorpos/efeitos dos fármacos , Dissulfetos/análise , Hemocianinas/química , Peptídeos/química , Sequência de Aminoácidos , Animais , Hemocianinas/farmacologia , Soros Imunes , Peptídeos/farmacologia , Relaxina/química , Estrelas-do-Mar
6.
Nat Immunol ; 20(5): 593-601, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30886417

RESUMO

Interferon-λ (IFN-λ) acts on mucosal epithelial cells and thereby confers direct antiviral protection. In contrast, the role of IFN-λ in adaptive immunity is far less clear. Here, we report that mice deficient in IFN-λ signaling exhibited impaired CD8+ T cell and antibody responses after infection with a live-attenuated influenza virus. Virus-induced release of IFN-λ triggered the synthesis of thymic stromal lymphopoietin (TSLP) by M cells in the upper airways that, in turn, stimulated migratory dendritic cells and boosted antigen-dependent germinal center reactions in draining lymph nodes. The IFN-λ-TSLP axis also boosted production of the immunoglobulins IgG1 and IgA after intranasal immunization with influenza virus subunit vaccines and improved survival of mice after challenge with virulent influenza viruses. IFN-λ did not influence the efficacy of vaccines applied by subcutaneous or intraperitoneal routes, indicating that IFN-λ plays a vital role in potentiating adaptive immune responses that initiate at mucosal surfaces.


Assuntos
Imunidade Adaptativa/imunologia , Citocinas/imunologia , Imunidade nas Mucosas/imunologia , Interleucinas/imunologia , Imunidade Adaptativa/efeitos dos fármacos , Imunidade Adaptativa/genética , Animais , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/virologia , Imunidade nas Mucosas/efeitos dos fármacos , Imunidade nas Mucosas/genética , Imunização/métodos , Vírus da Influenza A/efeitos dos fármacos , Vírus da Influenza A/imunologia , Vírus da Influenza A/fisiologia , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/imunologia , Interleucinas/administração & dosagem , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Knockout , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/virologia , Receptores de Interferon/genética , Receptores de Interferon/imunologia , Receptores de Interferon/metabolismo
7.
Methods Mol Biol ; 1943: 333-346, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30838627

RESUMO

For the systemic application of nucleic acids [plasmid DNA (pDNA) and small interfering RNA (siRNA)], safe and efficient carriers that overcome the poor pharmacokinetic properties of nucleic acids are required. A cationic liposome that can formulate lipoplexes with nucleic acids has significant promise as an efficient delivery system in gene therapy. To achieve in vivo stability and long circulation, most lipoplexes are modified with PEG (PEGylation). However, we reported that PEGylated liposomes lose their long-circulating properties when they are injected repeatedly at certain intervals in the same animal. This unexpected and undesirable phenomenon is referred to as the accelerated blood clearance (ABC) phenomenon. Anti-PEG IgM produced in response to the first dose of PEGylated liposomes has proven to be a major cause of the ABC phenomenon. Therefore, in a repeated dosing schedule, the detection of anti-PEG IgM in an animal treated with PEGylated lipoplex could be essential to predict the occurrence of the ABC phenomenon. This chapter introduces a method for the evaluation of serum anti-PEG IgM by a simple ELISA procedure, and describes some precautions associated with this method.


Assuntos
Técnicas de Transferência de Genes , Imunoglobulina M/sangue , Polietilenoglicóis/toxicidade , Animais , Formação de Anticorpos/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática/métodos , Terapia Genética/métodos , Imunoglobulina M/imunologia , Lipossomos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ácidos Nucleicos/genética , Polietilenoglicóis/administração & dosagem
8.
Eur J Pharmacol ; 851: 52-62, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30753864

RESUMO

Thymic stromal lymphopoietin (TSLP) is a key epithelial-derived factor that aggravates allergic diseases. Therefore, TSLP inhibitors are candidate compounds for the treatment of allergic diseases. Previously, we reported that KCMH-1, a mouse keratinocyte cell line, constitutively produces TSLP. In this study, we tried to identify inhibitors of TSLP by screening 2169 compounds in KCMH-1 cells and found one such chalcone derivative (code no. 16D10). 16D10 inhibited TSLP expression and TSLP promoter activation in HaCaT cells, a human keratinocyte cell line. Although nuclear factor kappa-B (NF-κB) is a key transcription factor for the induction of TSLP, 16D10 did not inhibit the activation pathway of NF-κB, such as degradation of inhibitor of κB (IκB) and p65 nuclear translocation. 16D10 activated the Kelch-like ECH-associated protein 1 (Keap1)-nuclear factor (erythroid-derived 2)-like 2 (Nrf2) system, although this system was not involved in the inhibitory effect of 16D10. 16D10 also inhibited TSLP production in a lipopolysaccharide (LPS)- or ovalbumin (OVA)-induced air-pouch-type inflammation model. Further, repeated 16D10 administration diminished serum immunoglobulin G1 (IgG1) and IgE concentration in an OVA-induced air-pouch-type sensitization model. Taken together, these results indicate that 16D10 is an inhibitor of TSLP production and has an anti-allergic effect. This inhibitory effect is independent of the activation of NF-κB and the Keap1-Nrf2 system. Therefore, 16D10 could be a new type of candidate drug for allergic diseases.


Assuntos
Formação de Anticorpos/efeitos dos fármacos , Chalconas/química , Chalconas/farmacologia , Citocinas/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Ovalbumina/farmacologia , Animais , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Humanos , Queratinócitos/imunologia , Masculino , Camundongos , NF-kappa B/metabolismo
9.
Int J Biol Macromol ; 125: 865-875, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30576729

RESUMO

Astragalus polysaccharides (APS) have long been well known as immune boosters, but have not been fully exploited in clinical settings. Here, poly(lactic-co-glycolic acid) (PLGA) was used to form a nanocarrier for APS to enhance its bioavailability. The aim was to improve the immunoadjuvanticity of conventional APS-loaded PLGA-based nanoparticles (NPs), referred to as APSPs, and to optimize the synthesis parameters to maximize the encapsulation efficiency (EE). As slow drug release can cause insufficient immune responses, ammonium bicarbonate was used to produce pH-responsive APSPs. The optimum parameters for maximizing EE (mean maximum experimental EE: 65.23 ±â€¯0.51%) were an oil phase (O)/internal aqueous phase (W1) ratio of 7:1, an external aqueous phase (W2)/preliminary emulsion (PE) ratio of 5:1, and a Pluronic F-68 concentration of 1.1%. Moreover, the pH-responsive APSPs had low cytotoxicity and significantly enhanced mice splenic lymphocyte proliferation. The increased T-cell CD4+/CD8+ ratio after pH-responsive APSP treatment of mice splenic lymphocytes compared with free APS, blank PLGA NP, and conventional APSP treatment demonstrated its excellent immunoadjuvanticity. This study provides abundant evidence that these novel PLGA-based pH-responsive NPs enhanced the immunoadjuvanticity of APS. Furthermore, pH-responsive APSPs synthesized using the optimum parameters exhibited long-term stability in normal storage conditions, suggesting suitability for clinical application.


Assuntos
Formação de Anticorpos/efeitos dos fármacos , Astrágalo (Planta)/química , Astrágalo (Planta)/imunologia , Nanopartículas/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Polissacarídeos/química , Polissacarídeos/imunologia , Animais , Proliferação de Células/efeitos dos fármacos , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacologia , Portadores de Fármacos/química , Emulsões/química , Concentração de Íons de Hidrogênio , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos ICR , Linfócitos T/efeitos dos fármacos
10.
J Infect Dis ; 219(2): 323-334, 2019 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-30289460

RESUMO

Protection against encapsulated bacteria can be elicited using polysaccharide vaccines. These antigens often behave as T-cell-independent type 2 antigens (TI-2 Ags). However, TI-2 Ags, including pneumococcal polysaccharides, often elicit weak immunoglobulin G (IgG) responses and are refractive to boosting. Conjugate vaccines have not completely overcome this challenge and hence, alternative strategies are required to enhance polysaccharide vaccine responses. Herein, we describe an adjuvant consisting of a Toll-like receptor and C-type lectin receptor agonist pairing that significantly increases primary immunoglobulin M and IgG responses to TI-2 Ags as well as enables significant boosting when coadministered with polysaccharide vaccines. Consistent with this, the adjuvant significantly increased the generation of both TI-2 memory B cells and long-lived antibody secreting cells. Adjuvant effects were highly dependent on B-cell-intrinsic MyD88, but not Trif expression. Importantly, coadministration of the adjuvant with the Pneumovax vaccine significantly increased the protective efficacy of vaccination in a lethal challenge mouse model of pneumococcal respiratory infection. Collectively, these data provide evidence that B-cell-directed adjuvants have promise in significantly improving the quality and quantity of serologic and B-cell memory responses to clinically relevant polysaccharide vaccines.


Assuntos
Adjuvantes Imunológicos/farmacologia , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Polissacarídeos/imunologia , Proteínas Adaptadoras de Transporte Vesicular/genética , Animais , Anticorpos Antibacterianos , Células Produtoras de Anticorpos/imunologia , Linfócitos B/imunologia , Vacinas Bacterianas/imunologia , Modelos Animais de Doenças , Feminino , Imunoglobulina G/imunologia , Imunoglobulina M , Lectinas Tipo C/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/genética , Infecções Pneumocócicas/imunologia , Vacinas Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Receptores Toll-Like/imunologia , Vacinação , Vacinas Conjugadas/imunologia
11.
Int J Nanomedicine ; 13: 6699-6715, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30425484

RESUMO

Background: Influenza (flu) is a constant threat to humans and animals, and vaccination is one of the most effective ways to mitigate the disease. Due to incomplete protection induced by current flu vaccines, development of novel flu vaccine candidates is warranted to achieve greater efficacy against constantly evolving flu viruses. Methods: In the present study, we used liposome nanoparticle (<200 nm diameter)-based subunit flu vaccine containing ten encapsulated highly conserved B and T cell epitope peptides to induce protective immune response against a zoonotic swine influenza A virus (SwIAV) H1N1 challenge infection in a pig model. Furthermore, we used monosodium urate (MSU) crystals as an adjuvant and co-administered the vaccine formulation as an intranasal mist to flu-free nursery pigs, twice at 3-week intervals. Results: Liposome peptides flu vaccine delivered with MSU adjuvant improved the hemagglutination inhibition antibody titer and mucosal IgA response against the SwIAV challenge and also against two other highly genetically variant IAVs. Liposomal vaccines also enhanced the frequency of peptides and virus-specific T-helper/memory cells and IFN-γ response. The improved specific cellular and mucosal humoral immune responses in adjuvanted liposomal peptides flu vaccine partially protected pigs from flu-induced fever and pneumonic lesions, and reduced the nasal virus shedding and viral load in the lungs. Conclusion: Overall, our study shows great promise for using liposome and MSU adjuvant- based subunit flu vaccine through the intranasal route, and provides scope for future, pre-clinical investigations in a pig model for developing potent human intranasal subunit flu vaccines.


Assuntos
Adjuvantes Imunológicos/farmacologia , Imunidade , Vacinas contra Influenza/imunologia , Nanopartículas/química , Infecções por Orthomyxoviridae/imunologia , Peptídeos/imunologia , Ácido Úrico/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Administração Intranasal , Sequência de Aminoácidos , Animais , Formação de Anticorpos/efeitos dos fármacos , Temperatura Corporal/efeitos dos fármacos , Citocinas/biossíntese , Cães , Imunidade/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Memória Imunológica/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N1 , Lipossomos , Pulmão/patologia , Pulmão/virologia , Células Madin Darby de Rim Canino , Nanopartículas/ultraestrutura , Infecções por Orthomyxoviridae/virologia , Peptídeos/química , Sus scrofa , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Vacinação , Carga Viral/efeitos dos fármacos
12.
Sci Rep ; 8(1): 16236, 2018 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-30390012

RESUMO

CpG-DNA activates various immune cells, contributing to the host defense against bacteria. Here, we examined the biological function of CpG-DNA in the production of bacteria-reactive antibodies. The administration of CpG-DNA increased survival in mice following infection with methicillin-resistant S. aureus and protected immune cell populations in the peritoneal cavity, bone marrow, and spleen. CpG-DNA injection likewise increased bacteria-reactive antibodies in the mouse peritoneal fluid and serum, which was dependent on TLR9. B cells isolated from the peritoneal cavity produced bacteria-reactive antibodies in vitro following CpG-DNA administration that enhanced the phagocytic activity of the peritoneal cells. The bacteria-reactive monoclonal antibody enhanced phagocytosis in vitro and protected mice after S. aureus infection. Therefore, we suggest that CpG-DNA enhances the antibacterial activity of the immune system by protecting immune cells and triggering the production of bacteria-reactive antibodies. Consequently, we believe that monoclonal antibodies could aid in the treatment of antibiotic-resistant bacterial infections.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Staphylococcus aureus Resistente à Meticilina/imunologia , Oligodesoxirribonucleotídeos/administração & dosagem , Infecções Estafilocócicas/terapia , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Anticorpos Antibacterianos/metabolismo , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Formação de Anticorpos/efeitos dos fármacos , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Linfócitos B/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Fagocitose/efeitos dos fármacos , Fagocitose/imunologia , Infecções Estafilocócicas/sangue , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Receptor Toll-Like 9/genética , Receptor Toll-Like 9/imunologia , Receptor Toll-Like 9/metabolismo , Resultado do Tratamento
13.
Endocrinology ; 159(11): 3834-3847, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30307543

RESUMO

The autoimmune response that characterizes type 1 diabetes (T1D) has no clear cause. Extracellular vesicles (EVs) play an important role in triggering the immune response in other contexts. Here, we propose a model by which EVs isolated from human islets stimulate proinflammatory immune responses and lead to peripheral blood mononuclear cell (PBMC) activation. We show that human islet EVs are internalized by monocytes and B cells and lead to an increase in T-helper 1, 2, and 17 cytokine expression, as well as T and B cell proliferation. Importantly, we demonstrate memory T and B cell activation by EVs selectively in PBMCs of patients with T1D. Additionally, human islet EVs induce an increase in antibodies against glutamic acid decarboxylase 65 (GAD65) in T1D PBMCs. Furthermore, pretreatment of T1D PBMCs with ibrutinib, an inhibitor of Bruton tyrosine kinase, dampens EV-induced memory B cell activation and GAD65 antibody production. Collectively, our findings indicate a role for human islet EVs in mediating activation of B and T cells and GAD65 autoantibody production.


Assuntos
Autoanticorpos/imunologia , Linfócitos B/imunologia , Citocinas/imunologia , Diabetes Mellitus Tipo 1/imunologia , Vesículas Extracelulares/imunologia , Glutamato Descarboxilase/imunologia , Ilhotas Pancreáticas/imunologia , Monócitos/imunologia , Linfócitos T/imunologia , Adulto , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Autoanticorpos/efeitos dos fármacos , Proliferação de Células , Feminino , Humanos , Memória Imunológica/imunologia , Leucócitos Mononucleares/imunologia , Ativação Linfocitária , Masculino , Pessoa de Meia-Idade , Inibidores de Proteínas Quinases/farmacologia , Pirazóis/farmacologia , Pirimidinas/farmacologia , Células Th1/imunologia , Células Th17/imunologia , Células Th2/imunologia , Adulto Jovem
14.
J Immunol Res ; 2018: 2505818, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30276218

RESUMO

B cells are pathogenic in various disease processes and therefore represent an interesting target for the development of novel immunosuppressants. In the search for new therapeutic molecules, we utilized an in vitro B cell activation assay with ODN2006-stimulated Namalwa cells to screen a chemical library of small molecules for B cell modulating effects. OSU-T315, described as an inhibitor of integrin-linked kinase (ILK), was hereby identified as a hit. On human and murine primary B cells, OSU-T315 potently suppressed the proliferation and the production of antibodies and cytokines upon stimulation, suggesting that ILK could be a promising target in the modulation of B cell activity. Mice with B cell-specific knockout of ILK were generated. Surprisingly, knockout of ILK in murine B cells did not affect B cell function as assessed by several in vivo and ex vivo B cell assays and did not alter the B cell immunosuppressive activity of OSU-T315. In conclusion, OSU-T315 displays potency as B cell modulator, probably through a mechanism of action independent of ILK, and might serve as lead drug molecule for the development of novel B cell-selective drugs.


Assuntos
Linfócitos B/efeitos dos fármacos , Imunossupressores/uso terapêutico , Piperazinas/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico , Pirazóis/uso terapêutico , Animais , Formação de Anticorpos/efeitos dos fármacos , Linfócitos B/fisiologia , Linhagem Celular , Proliferação de Células , Citocinas/metabolismo , Humanos , Imunomodulação , Imunossupressores/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Terapia de Alvo Molecular , Piperazinas/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/genética , Pirazóis/farmacologia
15.
Toxicon ; 153: 62-71, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30171931

RESUMO

This study aimed to evaluate the effect of dietary ochratoxin A (OA), in the presence and absence of L-carnitine (LC) and vitamin E (VE), on the humoral immune responses of White Leghorn cockerels (WLC). One-day old white male Leghorn chicks were divided into 12 groups, having 20 birds each and were offered ration contaminated with OA (1.0 or 2.0 mg/kg feed) alone and concurrently with LC (1.0 g/kg) and/or VE (0.2 g/kg), for 42 days. The humoral immune responses were accessed by lymphoproliferative response to avian tuberculin, in-vivo phagosomes activity to carbon particles and antibody response to the sheep red blood cells (SRBCs). The dietary addition of OA alone suppressed the humoral immune responses, however, the exposure of birds to 1.0 mg/kg OA in the presence of LC and/or VE showed a significant reduction in OA induced immunotoxicity. This protective response was absent in the birds fed 2.0 mg/kg OA in the presence and absence of LC and/or VE. Histopathological and morphometric examination of the bursa of Fabricius exhibited a decrease in the severity and frequency of OA induced lesions in the presence of dietary LC and/or VE. The use of LC and VE as dietary supplement, can effectively overcome OA (≤1.0 mg/kg) induced immunosuppression.


Assuntos
Carnitina/administração & dosagem , Galinhas/imunologia , Ocratoxinas/antagonistas & inibidores , Ocratoxinas/toxicidade , Vitamina E/administração & dosagem , Ração Animal , Animais , Formação de Anticorpos/efeitos dos fármacos , Bolsa de Fabricius/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Dieta/veterinária , Imunidade Humoral/efeitos dos fármacos , Masculino , Fagocitose/efeitos dos fármacos
16.
J Vet Sci ; 19(6): 817-826, 2018 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-30173497

RESUMO

The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.


Assuntos
Adjuvantes Imunológicos/farmacologia , Proteínas Aviárias/uso terapêutico , Bolsa de Fabricius/metabolismo , Vírus da Influenza A Subtipo H9N2 , Vacinas contra Influenza/farmacologia , Infecções por Orthomyxoviridae/prevenção & controle , Peptídeos/uso terapêutico , Adjuvantes Imunológicos/administração & dosagem , Animais , Anticorpos Antivirais/imunologia , Formação de Anticorpos/efeitos dos fármacos , Proteínas Aviárias/administração & dosagem , Quimioterapia Combinada , Feminino , Testes de Inibição da Hemaglutinação , Imunoglobulina G/imunologia , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Peptídeos/administração & dosagem
17.
Viral Immunol ; 31(7): 486-491, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30133352

RESUMO

Epstein-Barr virus (EBV) is a gamma-herpesvirus persisting mainly in human B lymphocytes. EBV reactivation induces host cells to differentiate into plasma cells and is related to autoimmune diseases. Graves' disease, an autoimmune hyperthyroidism, is caused by the thyrotropin receptor antibody (TRAb), which overstimulates thyroid stimulating hormone receptor. The disease occurs predominantly in women, which suggests involvement with estrogen. Graves' disease patients and healthy controls have EBV-infected lymphocytes with TRAb on the surface (TRAb(+)EBV(+) cells) in peripheral blood mononuclear cells (PBMCs). TRAb can be produced by reactivation of EBV in vitro, which is an alternative system of antibody production. In this study, we cultured PBMCs from Graves' disease patients and healthy controls with 0, 1, and 100 nM estradiol, corresponding to control, midluteal, and pregnancy levels, respectively, and analyzed the levels of TRAb, total-IgG, and total-IgM during EBV reactivation. We found that 1 nM estradiol increased TRAb levels and 100 nM estradiol slightly lowered them in both patients and controls. In patients, IgM production at 100 nM estradiol was significantly lower than that at 0 nM estradiol (p = 0.028). Estradiol increased the ratio of IgG production to immunoglobulin G (IgG) and immunoglobulin M (IgM) production (IgG/IgG + IgM), which suggested an increase in class switch recombination in the process of EBV reactivation-induced Ig production. Moreover, TRAb production was stimulated by a midluteal level of estradiol and was suppressed by a pregnancy level of estradiol in controls and patients. These results were consistent with premenstrual worsening and maternity improving of autoimmune diseases, including Graves' disease.


Assuntos
Formação de Anticorpos/efeitos dos fármacos , Estradiol/administração & dosagem , Doença de Graves/virologia , Herpesvirus Humano 4/imunologia , Receptores da Tireotropina/imunologia , Adulto , Autoanticorpos/sangue , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Ciclo Menstrual/efeitos dos fármacos , Pessoa de Meia-Idade , Gravidez , Estatísticas não Paramétricas
18.
Blood ; 132(11): 1193-1197, 2018 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-30064978

RESUMO

Von Willebrand factor (VWF) modulates factor VIII (FVIII) clearance and the anti-FVIII immune response. Despite the high affinity that defines the FVIII/VWF interaction, association/dissociation kinetics dictates 2% to 5% FVIII being present as free protein. To avoid free FVIII when studying the FVIII-VWF complex in vivo, we designed a FVIII-nanobody fusion protein, with the nanobody part being directed against VWF. This fusion protein, designated FVIII-KB013bv, had a 25-fold higher affinity compared with B-domainless FVIII (BDD-FVIII) for VWF. In vitro analysis revealed full cofactor activity in 1-stage clotting and chromogenic assays (activity/antigen ratio 1.0 ± 0.3 and 1.1 ± 0.3, respectively). In vivo, FVIII-013bv displayed a twofold increased mean residence time compared with BDD-FVIII (3.0 hours vs 1.6 hours). In a tail clip-bleeding assay performed 24 hours after FVIII infusion, blood loss was significantly reduced in mice receiving FVIII-KB013bv vs BDD-FVIII (15 ± 7 µL vs 194 ± 146 µL; P = .0043). Unexpectedly, when examining anti-FVIII antibody formation in FVIII-deficient mice, the immune-response toward FVIII-KB013bv was significantly reduced compared with BDD-FVIII (1/8 vs 14/16 mice produced anti-FVIII antibodies after treatment with FVIII-KB013bv and BDD-FVIII, respectively). Our data show that a stabilized interaction between FVIII and VWF is associated with a prolonged survival of FVIII and a reduced immune response against FVIII.


Assuntos
Formação de Anticorpos/efeitos dos fármacos , Autoanticorpos , Fator VIII , Proteínas Recombinantes de Fusão , Anticorpos de Domínio Único/farmacologia , Fator de von Willebrand , Animais , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Fator VIII/imunologia , Fator VIII/farmacocinética , Fator VIII/farmacologia , Camundongos , Camundongos Mutantes , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/farmacocinética , Proteínas Recombinantes de Fusão/farmacologia , Fator de von Willebrand/imunologia , Fator de von Willebrand/metabolismo
19.
J Control Release ; 285: 12-22, 2018 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-29964134

RESUMO

Nanocarriers based on inverse hexagonal liquid crystalline phases (hexosomes) show promising potential as vaccine delivery systems. Their unique internal structure, composed of both lipophilic domains and water-containing channels, renders them capable of accommodating immunopotentiating compounds and antigens. However, their adjuvant properties are poorly understood. We hypothesized that the supramolecular structure of the lyotropic liquid crystalline phase influences the immunostimulatory activity of lipid-based nanocarriers. To test this, hexosomes were designed containing the lipid phytantriol (Phy) and the immunopotentiator monomycoloyl glycerol-1 (MMG-1). Self-assembly of Phy and MMG-1 into nanocarriers featuring an internal hexagonal phase was confirmed by small-angle X-ray scattering and cryogenic transmission electron microscopy. The effect of the nanostructure on the adjuvant activity was studied by comparing the immunogenicity of Phy/MMG-1 hexosomes with MMG-1-containing lamellar liquid crystalline nanoparticles (liposomes, CAF04). The quality and magnitude of the elicited immune responses were determined after vaccination of CB6/F1 mice using the Chlamydia trachomatis major outer membrane protein (MOMP) as antigen. MMG-1-based hexosomes potentiated significantly stronger MOMP-specific humoral responses than CAF04 liposomes. The liposome-based vaccine formulation induced a much stronger MOMP-specific cell-mediated immune response compared to hexosome-adjuvanted MOMP, which elicited minimal MOMP-specific T-cell stimulation after vaccination. Hence, our data demonstrates that hexosomal and liposomal adjuvants activate the immune system via different mechanisms. Our work provides valuable insights into the adjuvant potential of hexosomes and emphasizes that engineering of the supramolecular structure can be used to design adjuvants with customized immunological properties.


Assuntos
Adjuvantes Imunológicos/farmacologia , Vacinas Bacterianas/farmacologia , Infecções por Chlamydia/prevenção & controle , Chlamydia trachomatis/imunologia , Álcoois Graxos/farmacologia , Monoglicerídeos/farmacologia , Porinas/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/química , Animais , Formação de Anticorpos/efeitos dos fármacos , Vacinas Bacterianas/administração & dosagem , Infecções por Chlamydia/imunologia , Portadores de Fármacos/química , Álcoois Graxos/administração & dosagem , Álcoois Graxos/química , Feminino , Cristais Líquidos/química , Camundongos , Monoglicerídeos/administração & dosagem , Monoglicerídeos/química , Nanopartículas/química , Porinas/administração & dosagem , Vacinação
20.
Bull Exp Biol Med ; 165(3): 368-372, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30003418

RESUMO

The glycolipid biosurfactant complex from actinobacterium Rhodococcus ruber IEGM 231 inhibits the innate and adaptive immunity parameters after intraperitoneal and intramuscular injection. Marked suppression of antibody production, bactericidal potential, and production of proinflammatory cytokines by peritoneal macrophages, detected in vivo, do not agree with the previously detected immunostimulatory activity of biosurfactants towards the immunocompetent cell cultures; this fact indicates an important role of the cell environment in the formation of immune response under the effect of bacterial glycolipids.


Assuntos
Glicolipídeos/farmacologia , Imunidade Humoral/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , Rhodococcus/química , Tensoativos/farmacologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Contagem de Células , Relação Dose-Resposta Imunológica , Glicolipídeos/isolamento & purificação , Injeções Intramusculares , Injeções Intravenosas , Interleucina-1beta/biossíntese , Interleucina-1beta/imunologia , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Masculino , Camundongos , Cultura Primária de Células , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Tensoativos/isolamento & purificação , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologia , Zimosan/farmacologia
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