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2.
Infect Immun ; 87(12)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31548320

RESUMO

Lipopolysaccharides (LPSs) of Gram-negative bacteria comprise lipid A, core, and O-polysaccharide (OPS) components. Studies have demonstrated that LPSs isolated from the pathogenic species Burkholderia pseudomallei and Burkholderia mallei and from less-pathogenic species, such as Burkholderia thailandensis, are potent immune stimulators. The LPS structure of B. pseudomallei, the causative agent of melioidosis, is highly conserved in isolates from Thailand; however, the LPSs isolated from other, related species have not been characterized to enable understanding of their immune recognition and antigenicities. Here, we describe the structural and immunological characteristics of the LPSs isolated from eight Burkholderia species and compare those for B. pseudomallei to those for the other seven species. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), gas chromatography (GC), SDS-PAGE, Toll-like receptor 4 (TLR4) stimulation, and immunoblot analysis were performed on these Burkholderia species. MALDI-TOF profiles demonstrated that Burkholderia lipid A contains predominantly penta-acylated species modified with 4-amino-4-deoxy-arabinose residues at both terminal phosphate groups. The lipid A could be differentiated based on mass differences at m/z 1,511, 1,642, 1,773, and 1,926 and on fatty acid composition. LPSs of all species induced TLR4-dependent NF-κB responses; however, while SDS-PAGE analysis showed similar LPS ladder patterns for B. pseudomallei, B. thailandensis, and B. mallei, these patterns differed from those of other Burkholderia species. Interestingly, immunoblot analysis demonstrated that melioidosis patient sera cross-reacted with OPSs of other Burkholderia species. These findings can be used to better understand the characteristics of LPS in Burkholderia species, and they have implications for serological diagnostics based on the detection of antibodies to OPS.


Assuntos
Burkholderia mallei/imunologia , Burkholderia pseudomallei/imunologia , Burkholderia/imunologia , Lipídeo A/imunologia , Receptor 4 Toll-Like/metabolismo , Amino Açúcares/química , Anticorpos Antibacterianos/imunologia , Reações Cruzadas/imunologia , Humanos , Lipídeo A/química , Melioidose/imunologia , Melioidose/microbiologia , Conformação Molecular , Polissacarídeos Bacterianos/imunologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
3.
Infect Immun ; 87(12)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31527122

RESUMO

We previously demonstrated that recombinant protein PAc could be administered as an anticaries vaccine. However, the relatively weak immunogenicity of PAc limits its application. In the present study, we investigated the effect of two adjuvant combinations of chitosan plus Pam3CSK4 (chitosan-Pam3CSK4) and of chitosan plus monophosphoryl lipid A (chitosan-MPL) in the immune responses to the PAc protein in vivo and in vitro PAc-chitosan-Pam3CSK4 or PAc-chitosan-MPL promoted significantly higher PAc-specific antibody titers in serum and saliva, inhibited Streptococcus mutans colonization onto the tooth surfaces, and endowed better protection effect with significantly less caries activities than PAc alone. Chitosan-Pam3CSK4 and chitosan-MPL showed no statistically significant differences. In conclusion, our study demonstrated that the chitosan-Pam3CSK4 and chitosan-MPL combinations are promising for anticaries vaccine development.


Assuntos
Vacinas Bacterianas/imunologia , Quitosana/farmacologia , Cárie Dentária/prevenção & controle , Lipídeo A/análogos & derivados , Lipopeptídeos/farmacologia , Streptococcus mutans/imunologia , Adjuvantes Imunológicos , Animais , Cárie Dentária/microbiologia , Feminino , Imunogenicidade da Vacina/imunologia , Imunoglobulina A Secretora/análise , Lipídeo A/imunologia , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas NLR/agonistas , Streptococcus mutans/patogenicidade , Receptores Toll-Like/agonistas , Vacinas Sintéticas/imunologia , Fatores de Virulência/imunologia
4.
J Immunol Res ; 2019: 3974127, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31205956

RESUMO

Adjuvants are a diverse family of substances whose main objective is to increase the strength, quality, and duration of the immune response caused by vaccines. The most commonly used adjuvants are aluminum-based, oil-water emulsion, and bacterial-origin adjuvants. In this paper, we will discuss how the election of adjuvants is important for the adjuvant-mediated induction of immunity for different types of vaccines. Aluminum-based adjuvants are the most commonly used, the safest, and have the best efficacy, due to the triggering of a strong humoral response, albeit generating a weak induction of cell-mediated immune response. Freund's adjuvant is the most widely used oil-water emulsion adjuvant in animal trials; it stimulates inflammation and causes aggregation and precipitation of soluble protein antigens that facilitate the uptake by antigen-presenting cells (APCs). Adjuvants of bacterial origin, such as flagellin, E. coli membranes, and monophosphoryl lipid A (MLA), are known to potentiate immune responses, but their safety and risks are the main concern of their clinical use. This minireview summarizes the mechanisms that classic and novel adjuvants produce to stimulate immune responses.


Assuntos
Adjuvantes Imunológicos , Hidróxido de Alumínio/imunologia , Antígenos de Bactérias/imunologia , Emulsões , Escherichia coli/imunologia , Lipídeo A/análogos & derivados , Óleos , Animais , Humanos , Imunidade Celular , Lipídeo A/imunologia
5.
Cancer Immunol Immunother ; 68(7): 1211-1222, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31069460

RESUMO

Human tumor cells express antigens that serve as targets for the host cellular immune system. This phase 1 dose-escalating study was conducted to assess safety and tolerability of G305, a recombinant NY-ESO-1 protein vaccine mixed with glucopyranosyl lipid A (GLA), a synthetic TLR4 agonist adjuvant, in a stable emulsion (SE). Twelve patients with solid tumors expressing NY-ESO-1 were treated using a 3 + 3 design. The NY-ESO-1 dose was fixed at 250 µg, while GLA-SE was increased from 2 to 10 µg. Safety, immunogenicity, and clinical responses were assessed prior to, during, and at the end of therapy. G305 was safe and immunogenic at all doses. All related AEs were Grade 1 or 2, with injection site soreness as the most commonly reported event (100%). Overall, 75% of patients developed antibody response to NY-ESO-1, including six patients with increased antibody titer ( ≥ 4-fold rise) and three patients with seroconversion from negative (titer < 100) to positive (titer ≥ 100). CD4 T-cell responses were observed in 44.4% of patients; 33.3% were new responses and 1 was boosted ( ≥ 2-fold rise). Following treatment, 8 of 12 patients had stable disease for 3 months or more; at the end of 1 year, three patients had stable disease and nine patients were alive. G305 is a potent immunotherapeutic agent that can stimulate NY-ESO-1-specific antibody and T-cell responses. The vaccine was safe at all doses of GLA-SE (2-10 µg) and showed potential clinical benefit in this population of patients.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Antígenos de Neoplasias/administração & dosagem , Vacinas Anticâncer/administração & dosagem , Glucosídeos/administração & dosagem , Lipídeo A/administração & dosagem , Proteínas de Membrana/administração & dosagem , Neoplasias/terapia , Adjuvantes Imunológicos/efeitos adversos , Adulto , Idoso , Antígenos de Neoplasias/efeitos adversos , Antígenos de Neoplasias/imunologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Vacinas Anticâncer/efeitos adversos , Vacinas Anticâncer/imunologia , Drogas em Investigação/administração & dosagem , Drogas em Investigação/efeitos adversos , Feminino , Glucosídeos/efeitos adversos , Glucosídeos/imunologia , Humanos , Imunogenicidade da Vacina , Injeções Intramusculares , Lipídeo A/efeitos adversos , Lipídeo A/imunologia , Masculino , Proteínas de Membrana/efeitos adversos , Proteínas de Membrana/imunologia , Pessoa de Meia-Idade , Neoplasias/imunologia , Neoplasias/patologia , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/imunologia , Receptor 4 Toll-Like/agonistas , Receptor 4 Toll-Like/imunologia , Resultado do Tratamento , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/efeitos adversos , Vacinas Sintéticas/imunologia , Adulto Jovem
6.
Mol Immunol ; 111: 43-52, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30959420

RESUMO

Salmonella enterica serovar Typhimurium (S. Typhimurium) changes the structure of its lipopolysaccharide (LPS) in response to the environment. The two main LPS variants found in S. Typhimurium correspond to LPS with a hepta-acylated lipid A (LPS 430) and LPS with modified phosphate groups on its lipid A (LPS 435). We have previously shown that these modified LPS have a lower capacity than wild type (WT) LPS to induce the production of pro-inflammatory cytokines in mice. Nevertheless, it is not know if LPS 430 and LPS 435 could also subvert the innate immune responses in human cells. In this study, we found that LPS 430 and LPS 435 were less efficient than WT LPS to induce the production of pro-inflammatory cytokines by human monocytes, in addition we found a decreased dimerization of the TLR4/MD-2 complex in response to LPS 430, suggesting that structurally modified LPS are sensed differently than WT LPS by this receptor; however, LPS 430 and 435 induced similar activation of the transcription factors NF-κB p65, IRF3, p38 and ERK1/2 than WT LPS. Microarray analysis of LPS 430- and LPS 435-activated monocytes revealed a gene transcription profile with differences only in the expression levels of microRNA genes compared to the profile induced by WT LPS, suggesting that the lipid A modifications present in LPS 430 and LPS 435 have a moderate effect on the activation of the human TLR4/MD-2 complex. Our results are relevant to understand LPS modulation of immune responses and this knowledge could be useful for the development of novel adjuvants and immunomodulators.


Assuntos
Citocinas/imunologia , Inflamação/imunologia , Lipopolissacarídeos/imunologia , Antígeno 96 de Linfócito/imunologia , Monócitos/imunologia , Salmonella typhimurium/imunologia , Receptor 4 Toll-Like/imunologia , Acilação/imunologia , Dimerização , Humanos , Inflamação/microbiologia , Lipídeo A/imunologia , Monócitos/microbiologia , Infecções por Salmonella/imunologia , Infecções por Salmonella/microbiologia , Transdução de Sinais/imunologia , Fatores de Transcrição/imunologia , Transcrição Genética/imunologia
7.
J Biol Chem ; 294(22): 8872-8884, 2019 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-31000631

RESUMO

Receptor-interacting protein kinase 3 (RIPK3) is a key regulator of programmed cell death and inflammation during viral infection or sterile tissue injury. Whether and how bacterial infection also activates RIPK3-dependent immune responses remains poorly understood. Here we show that bacterial lipids (lipid IVa or lipid A) form a complex with high mobility group box 1 (HMGB1), released by activated immune cells or damaged tissue during bacterial infection, and that this complex triggers RIPK3- and TIR domain-containing adapter-inducing IFN-ß (TRIF)-dependent immune responses. We found that these responses lead to macrophage death, interleukin (IL)-1α release, and IL-1ß maturation. In an air-pouch inflammatory infiltration model, genetic deletion of Ripk3, Trif, or IL-1 receptor (Il-1R), or monoclonal antibody-mediated HMGB1 neutralization uniformly attenuated inflammatory responses induced by Gram-negative bacteria that release lipid IVa and lipid A. These findings uncover a previously unrecognized mechanism by which host factors and bacterial components work in concert to orchestrate immune responses.


Assuntos
Apoptose , Proteína HMGB1/metabolismo , Lipídeo A/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Glicolipídeos/imunologia , Glicolipídeos/metabolismo , Bactérias Gram-Negativas/metabolismo , Proteína HMGB1/imunologia , Humanos , Inflamação/metabolismo , Inflamação/patologia , Interleucina-1alfa/metabolismo , Interleucina-1beta/metabolismo , Lipídeo A/análogos & derivados , Lipídeo A/imunologia , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Ligação Proteica , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo
8.
Innate Immun ; 25(3): 203-212, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30894093

RESUMO

Endotoxin research in recent years at the molecular level has required chemically synthesized lipid A without contamination by other bioactive components. Total synthesis of Escherichia coli-type lipid A was achieved in the 1980s by the challenging spirits of the scientists at Osaka University, Japan. They clarified the role of lipid A in the immunological activities of endotoxin in collaboration with Japanese and German researchers, based on the friendships that existed between them. This article introduces the great contributions made by three generations of professors, Tetsuo Shiba, Shoichi Kusumoto, and Koichi Fukase, at the Laboratory of Natural Product Chemistry at Osaka University, to the study over four decades of endotoxin.


Assuntos
Química Orgânica/história , Escherichia coli/metabolismo , Lipídeo A/síntese química , Choque Séptico/imunologia , História do Século XX , História do Século XXI , Cooperação Internacional , Japão , Lipídeo A/imunologia
9.
Pathog Dis ; 76(8)2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30476069

RESUMO

Bacterial infection of the kidney leads to a rapid cascade of host protective responses, many of which are still poorly understood. We have previously shown that following kidney infection with uropathogenic Escherichia coli (UPEC), vascular coagulation is quickly initiated in local perivascular capillaries that protects the host from progressing from a local infection to systemic sepsis. The signaling mechanisms behind this response have not however been described. In this study, we use a number of in vitro and in vivo techniques, including intravital microscopy, to identify two previously unrecognized components influencing this protective coagulation response. The acylation state of the Lipid A of UPEC lipopolysaccharide (LPS) is shown to alter the kinetics of local coagulation onset in vivo. We also identify epithelial CD147 as a potential host factor influencing infection-mediated coagulation. CD147 is expressed by renal proximal epithelial cells infected with UPEC, contingent to bacterial expression of the α-hemolysin toxin. The epithelial CD147 subsequently can activate tissue factor on endothelial cells, a primary step in the coagulation cascade. This study emphasizes the rapid, multifaceted response of the kidney tissue to bacterial infection and the interplay between host and pathogen during the early hours of renal infection.


Assuntos
Infecções Bacterianas/sangue , Infecções Bacterianas/imunologia , Infecções Bacterianas/metabolismo , Basigina/metabolismo , Coagulação Sanguínea , Lipídeo A/imunologia , Nefrite/etiologia , Nefrite/metabolismo , Animais , Biomarcadores , Linhagem Celular , Citocinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Proteoma , Proteômica/métodos , Ratos , Transdução de Sinais
10.
PLoS One ; 13(11): e0206838, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30496299

RESUMO

BACKGROUND: We evaluated the safety and immunogenicity of (i) an intradermal HIV-DNA regimen given with/without intradermal electroporation (EP) as prime and (ii) the impact of boosting with modified vaccinia virus Ankara (HIV-MVA) administered with or without subtype C CN54rgp140 envelope protein adjuvanted with Glucopyranosyl Lipid A (GLA-AF) in volunteers from Tanzania and Mozambique. METHODS: Healthy HIV-uninfected adults (N = 191) were randomized twice; first to one of three HIV-DNA intradermal priming regimens by needle-free ZetaJet device at weeks 0, 4 and 12 (Group I: 2x0.1mL [3mg/mL], Group II: 2x0.1mL [3mg/mL] plus EP, Group III: 1x0.1mL [6mg/mL] plus EP). Second the same volunteers received 108 pfu HIV-MVA twice, alone or combined with CN54rgp140/GLA-AF, intramuscularly by syringe, 16 weeks apart. Additionally, 20 volunteers received saline placebo. RESULTS: Vaccinations and electroporation did not raise safety concerns. After the last vaccination, the overall IFN-γ ELISpot response rate to either Gag or Env was 97%. Intradermal electroporation significantly increased ELISpot response rates to HIV-DNA-specific Gag (66% group I vs. 86% group II, p = 0.026), but not to the HIV-MVA vaccine-specific Gag or Env peptide pools nor the magnitude of responses. Co-administration of rgp140/GLA-AF with HIV-MVA did not impact the frequency of binding antibody responses against subtype B gp160, C gp140 or E gp120 antigens (95%, 99%, 79%, respectively), but significantly enhanced the magnitude against subtype B gp160 (2700 versus 300, p<0.001) and subtype C gp140 (24300 versus 2700, p<0.001) Env protein. At relatively low titers, neutralizing antibody responses using the TZM-bl assay were more frequent in vaccinees given adjuvanted protein boost. CONCLUSION: Intradermal electroporation increased DNA-induced Gag response rates but did not show an impact on Env-specific responses nor on the magnitude of responses. Co-administration of HIV-MVA with rgp140/GLA-AF significantly enhanced antibody responses.


Assuntos
Vacinas contra a AIDS/imunologia , Infecções por HIV/prevenção & controle , Imunogenicidade da Vacina , Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Vacinas contra a AIDS/administração & dosagem , Vacinas contra a AIDS/efeitos adversos , Vacinas contra a AIDS/genética , Administração Cutânea , Adulto , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Eletroporação , Feminino , Glucosídeos/imunologia , Anticorpos Anti-HIV/sangue , Anticorpos Anti-HIV/imunologia , HIV-1/imunologia , Voluntários Saudáveis , Humanos , Imunização Secundária/métodos , Lipídeo A/imunologia , Masculino , Moçambique , Tanzânia , Vacinação/métodos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/efeitos adversos , Vacinas de DNA/genética , Vírus Vaccinia/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/efeitos adversos , Vacinas Virais/genética , Adulto Jovem , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia
11.
Macromol Biosci ; 18(12): e1800301, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30407735

RESUMO

Exosomes (EXO) are considered to be versatile carriers for biomolecules; however, the delivery of therapeutic peptides using EXOs poses several challenges. In this study, the efficiency of serum-derived EXOs in delivering tyrosinase-related protein-2 (TRP2) peptides to lymph nodes is determined. TRP2 peptides are successfully incorporated into EXOs, which show a uniform and narrow size distribution of around 45 nm. The TRP2-incorporated exosomes (EXO-TRP2) are efficiently internalized into macrophages and dendritic cells, and are seen to display a punctate distribution. EXOs loaded with TRP2 together with MPLA, (EXO-MPLA-TRP2) result in a strong release of proinflammatory cytokines (TNF-α and IL-6) from both RAW264.7 and DC2.4 cells. Finally, subcutaneous injection of fluorescently labeled EXO-TRP2 followed by ex vivo imaging using in vivo imaging system (IVIS) show a strong fluorescent signal in the lymph nodes after only 1 h, which is maintained until at least 4 h after injection. Taken together, the findings suggest that serum-derived EXOs can serve as promising carriers to deliver therapeutic peptides to lymph nodes for immunotherapy.


Assuntos
Adjuvantes Imunológicos/farmacocinética , Sistemas de Liberação de Medicamentos/métodos , Exossomos/metabolismo , Lipídeo A/análogos & derivados , Linfonodos/efeitos dos fármacos , Proteínas de Membrana/farmacocinética , Fragmentos de Peptídeos/farmacocinética , Adjuvantes Imunológicos/química , Animais , Transporte Biológico , Linhagem Celular , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Composição de Medicamentos/métodos , Eletroporação/métodos , Exossomos/química , Exossomos/transplante , Corantes Fluorescentes/farmacocinética , Expressão Gênica , Injeções Subcutâneas , Interleucina-6/genética , Interleucina-6/imunologia , Lipídeo A/química , Lipídeo A/imunologia , Lipídeo A/farmacocinética , Linfonodos/citologia , Linfonodos/imunologia , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Camundongos , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/imunologia , Células RAW 264.7 , Rodaminas/farmacocinética , Saponinas/química , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
12.
Drugs Aging ; 35(12): 1031-1040, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30370455

RESUMO

Shingrix® is a recombinant zoster vaccine (RZV) that contains a varicella zoster virus glycoprotein E antigen and the AS01B adjuvant system. The subunit vaccine is approved for the prevention of herpes zoster (HZ) [EU, USA, Japan, Canada and Australia] and postherpetic neuralgia (PHN) [EU and Australia] in adults aged ≥ 50 years. In the pivotal trials in adults aged ≥ 50 years (ZOE-50) and ≥ 70 years (ZOE-70), RZV significantly reduced the risk of HZ and PHN. Its protective efficacy waned minimally over 4 years and was well preserved in adults aged ≥ 70 years. In patients with breakthrough disease, RZV reduced HZ-related pain severity, burden of illness and burden of interference with activities of daily living. RZV was more reactogenic than placebo, with injection-site reactions, myalgia and fatigue being the most common solicited adverse reactions. However, most solicited adverse reactions were transient and were mild to moderate in severity. RZV represents a novel, highly effective and well-tolerated vaccine option for HZ and PHN in adults aged ≥ 50 years. RZV is not contraindicated in immunocompromised individuals, and is preferred over a live attenuated HZ vaccine in immunocompetent individuals, according to the US and Canadian guidelines.


Assuntos
Vacina contra Herpes Zoster/administração & dosagem , Herpes Zoster/prevenção & controle , Herpesvirus Humano 3/imunologia , Atividades Cotidianas , Idoso , Humanos , Hospedeiro Imunocomprometido , Lipídeo A/análogos & derivados , Lipídeo A/imunologia , Pessoa de Meia-Idade , Neuralgia Pós-Herpética/prevenção & controle , Saponinas/imunologia , Vacinas de Subunidades/administração & dosagem , Proteínas do Envelope Viral/imunologia
13.
Sci Adv ; 4(9): eaas9930, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30221194

RESUMO

Adjuvants are key to shaping the immune response to vaccination, but to date, no adjuvant suitable for human use has been developed for intradermal vaccines. These vaccines could be self-administered and sent through the mail as they do not require long needles or technical expertise in immunization. In the event of a pandemic outbreak, this approach could alleviate the congregation of patients in health centers and thus reduce the potential of these centers to enhance the spread of lethal infection. A reliable and potent vaccine system for self-administration would provide an effective countermeasure for delivery through existing product distribution infrastructure. We report results from preclinical and clinical trials that demonstrate the feasibility of an adjuvanted, intradermal vaccine that induced single shot protection in ferrets and seroprotection in humans against one of the more lethal strains of pandemic flu, Indonesia H5N1. In the human trial, the vaccine was safe and clinical responses were above approvable endpoints for a protective flu vaccine. Inclusion of a modern TLR4 (Toll-like receptor 4) agonist-based adjuvant was critical to the development of the response in the intradermal groups. In humans, this is the first report of a safe and effective intradermal adjuvant, GLA-AF (aqueous formulation of glucopyranosyl lipid adjuvant), and provides a future path for developing a vaccine-device combination for distribution by mail and self-administration in case of a pandemic.


Assuntos
1,2-Dipalmitoilfosfatidilcolina/imunologia , Adjuvantes Imunológicos/farmacologia , Vacinas contra Influenza/farmacologia , Lipídeo A/análogos & derivados , 1,2-Dipalmitoilfosfatidilcolina/efeitos adversos , 1,2-Dipalmitoilfosfatidilcolina/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/efeitos adversos , Adulto , Animais , Combinação de Medicamentos , Feminino , Furões , Cobaias , Humanos , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/imunologia , Virus da Influenza A Subtipo H5N1/patogenicidade , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/efeitos adversos , Injeções Intradérmicas , Lipídeo A/efeitos adversos , Lipídeo A/imunologia , Lipídeo A/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Receptor 4 Toll-Like/agonistas
14.
PLoS One ; 13(9): e0204491, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30248142

RESUMO

One strategy to control leishmaniasis is vaccination with potent antigens alongside suitable adjuvants. The use of toll-like receptor (TLR) agonists as adjuvants is a promising approach in Leishmania vaccine research. Leishmania (L.) tropica is among the less-investigated Leishmania species and a causative agent of cutaneous and sometimes visceral leishmaniasis with no approved vaccine against it. In the present study, we assessed the adjuvant effects of a TLR4 agonist, monophosphoryl lipid A (MPL) and a TLR7/8 agonist, R848 beside two different types of Leishmania vaccine candidates; namely, whole-cell soluble L. tropica antigen (SLA) and recombinant L. tropica stress-inducible protein-1 (LtSTI1). BALB/c mice were vaccinated three times by the antigens (SLA or LtSTI1) with MPL or R848 and then were challenged by L. tropica. Delayed-type hypersensitivity (DTH), parasite load, disease progression and cytokines (IL-10 and IFN-γ) responses were assessed. In general compared to SLA, application of LtSTI1 resulted in higher DTH, higher IFN-γ response and lower lymph node parasite load. Also compared to R848, MPL as an adjuvant resulted in higher DTH and lower lymph node parasite load. Although, no outstanding ability for SLA and R848 in evoking immune responses of BALB/c mice against L. tropica infection could be observed, our data suggest that LtSTI1 and MPL have a better potential to control L. tropica infection and could be pursued for the development of effective vaccination strategies.


Assuntos
Adjuvantes Imunológicos , Imidazóis/imunologia , Leishmania tropica/imunologia , Vacinas contra Leishmaniose/imunologia , Leishmaniose/prevenção & controle , Lipídeo A/análogos & derivados , Adjuvantes Imunológicos/administração & dosagem , Animais , Antígenos de Protozoários/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Imidazóis/administração & dosagem , Leishmaniose/imunologia , Vacinas contra Leishmaniose/administração & dosagem , Lipídeo A/administração & dosagem , Lipídeo A/imunologia , Camundongos Endogâmicos BALB C , Carga Parasitária , Proteínas de Protozoários/imunologia , Distribuição Aleatória , Proteínas Recombinantes/imunologia , Receptores Toll-Like/agonistas , Vacinação , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologia
15.
Int J Pharm ; 549(1-2): 404-414, 2018 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-30075250

RESUMO

This study reports on the immunogenicity assessment of a novel chimeric peptide vaccine including Tax, gp21, gp46, and gag immunodominant epitopes of human T-cell lymphotropic virus type 1 (HTLV-1) to induce immunity against HTLV-1 after subcutaneous (SC) or intranasal administration in a mice model. Additionally, to elevate the efficacy of the HTLV-1 vaccine, the chimera was physically mixed with monophosphoryl lipid A (MPLA) or ISCOMATRIX (IMX) adjuvants. For this purpose, the ISCOMATRIX with a size range of 40-60 nm were prepared using lipid film hydration method. Our investigation revealed that the mixture of IMX and chimera could significantly increase antibody titers containing IgG2a, and mucosal IgA, as well as IFN-γ and IL-10 cytokines and decrease the level of TGF-ß1, compared to other vaccine formulations. The intranasal delivery of chimera vaccine in the absence or presence adjuvants stimulated potent mucosal sIgA titer relative to subcutaneous immunization. Furthermore, the SC or nasal delivery of various vaccine formulations could shift the immunity toward cell-mediated responses, as evident by higher IgG2a and IFN-γ, as well as suppressed TGF-ß1 level. Our findings suggest that proper design, construction, and immunization of multi-epitope vaccine are essential for developing an effective HTLV-1 vaccine.


Assuntos
Antígenos HTLV-I/administração & dosagem , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Imunidade nas Mucosas , Imunogenicidade da Vacina , Mucosa Nasal/imunologia , Potência de Vacina , Vacinas Virais/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Administração Intranasal , Animais , Anticorpos Antivirais/sangue , Colesterol/administração & dosagem , Colesterol/imunologia , Citocinas/sangue , Combinação de Medicamentos , Antígenos HTLV-I/imunologia , Imunidade Celular , Imunização , Epitopos Imunodominantes , Injeções Subcutâneas , Lipídeo A/administração & dosagem , Lipídeo A/análogos & derivados , Lipídeo A/imunologia , Masculino , Camundongos Endogâmicos BALB C , Mucosa Nasal/virologia , Fosfolipídeos/administração & dosagem , Fosfolipídeos/imunologia , Saponinas/administração & dosagem , Saponinas/imunologia , Vacinas de Subunidades/administração & dosagem , Vacinas de Subunidades/imunologia , Vacinas Virais/imunologia
16.
Front Immunol ; 9: 1888, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30154796

RESUMO

Lipopolysaccharides (LPS) are potent activator of the innate immune response through the binding to the myeloid differentiation protein-2 (MD-2)/toll-like receptor 4 (TLR4) receptor complexes. Although a variety of LPSs have been characterized so far, a detailed molecular description of the structure-activity relationship of the lipid A part has yet to be clarified. Photosynthetic Bradyrhizobium strains, symbiont of Aeschynomene legumes, express distinctive LPSs bearing very long-chain fatty acids with a hopanoid moiety covalently linked to the lipid A region. Here, we investigated the immunological properties of LPSs isolated from Bradyrhizobium strains on both murine and human immune systems. We found that they exhibit a weak agonistic activity and, more interestingly, a potent inhibitory effect on MD-2/TLR4 activation exerted by toxic enterobacterial LPSs. By applying computational modeling techniques, we also furnished a plausible explanation for the Bradyrhizobium LPS inhibitory activity at atomic level, revealing that its uncommon lipid A chemical features could impair the proper formation of the receptorial complex, and/or has a destabilizing effect on the pre-assembled complex itself.


Assuntos
Bradyrhizobium/imunologia , Lipídeo A/imunologia , Animais , Linhagem Celular , Citocinas/metabolismo , Feminino , Humanos , Imunidade Inata , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Lipídeo A/química , Lipídeo A/metabolismo , Antígeno 96 de Linfócito/química , Antígeno 96 de Linfócito/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , Conformação Molecular , Ligação Proteica , Relação Estrutura-Atividade , Receptor 4 Toll-Like/química , Receptor 4 Toll-Like/metabolismo
17.
J Investig Med ; 66(8): 1124-1132, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-29997146

RESUMO

Sm-p80, the large subunit of Schistosoma mansoni calpain, is a leading candidate for a schistosomiasis vaccine. The prophylactic and antifecundity efficacy of Sm-p80 has been tested in three animal models (mouse, hamster and baboon) using a multitude of vaccine formulations and approaches. In our continual effort to enhance the vaccine efficacy, in this study, we have utilized the adjuvant, synthetic hexa-acylated lipid A derivative, glucopyranosyl lipid A (GLA) formulated in aluminum (GLA-Alum) with recombinant Sm-p80. The rSm-p80+GLA-Alum immunization regimen provided 33.33%-53.13% reduction in worm burden in the mouse model and 38% worm burden reduction in vaccinated baboons. Robust Sm-p80-specific immunoglobulin (Ig)G, IgG1, IgG2a and IgM responses were observed in all immunized animals. The rSm-p80+GLA-Alum coadministration induced a mix of T-helper (Th) cells (Th1, Th2 and Th17) responses as determined via the release of interleukin (IL)-2, IL-4, IL-18, IL-21, IL-22 and interferon-γ.


Assuntos
Adjuvantes Imunológicos/farmacologia , Compostos de Alúmen/farmacologia , Antígenos de Helmintos/imunologia , Glucosídeos/imunologia , Lipídeo A/imunologia , Schistosoma mansoni/imunologia , Receptor 4 Toll-Like/agonistas , Vacinas/imunologia , Animais , Proliferação de Células , Citocinas/biossíntese , Citocinas/genética , Feminino , Imunidade Humoral , Camundongos Endogâmicos C57BL , Papio , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismo , Esquistossomose mansoni/imunologia , Esquistossomose mansoni/parasitologia , Linfócitos T Auxiliares-Indutores/imunologia , Vacinação
18.
Sci Rep ; 8(1): 7096, 2018 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-29728623

RESUMO

We deciphered the mechanisms of production of pro- and anti-inflammatory cytokines by adherent human blood mononuclear cells (PBMC) activated by lipopolysaccharide (LPS) or monophosphoryl lipid A (MPLA). Both LPS and MPLA induced tumor necrosis factor (TNF) production proved to be dependent on the production of interleukin-1ß (IL-1ß). Of note, MPLA induced IL-1ß release in human adherent PBMCs whereas MPLA was previously reported to not induce this cytokine in murine cells. Both LPS and MPLA stimulatory effects were inhibited by Toll-like receptor-4 (TLR4) antagonists. Only monocytes activation by LPS was dependent on CD14. Other differences were noticed between LPS and MPLA. Among the different donors, a strong correlation existed in terms of the levels of TNF induced by different LPSs. In contrast, there was no correlation between the TNF productions induced by LPS and those induced by MPLA. However, there was a strong correlation when IL-6 production was analyzed. Blocking actin polymerization and internalization of the agonists inhibited MPLA induced TNF production while the effect on LPS induced TNF production depended on the donors (i.e. high TNF producers versus low TNF producers). Finally, conventional LPS, tolerized adherent PBMCs to TLR2 agonists, while MPLA primed cells to further challenge with TLR2 agonists.


Assuntos
Lipídeo A/análogos & derivados , Monócitos/imunologia , Antígenos de Bactérias/imunologia , Citocinas/biossíntese , Endocitose , Humanos , Tolerância Imunológica , Mediadores da Inflamação/metabolismo , Ligantes , Lipídeo A/imunologia , Receptores de Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/imunologia , Monócitos/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo
19.
J Immunol ; 201(1): 98-112, 2018 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-29769270

RESUMO

The involvement of innate receptors that recognize pathogen- and danger-associated molecular patterns is critical to programming an effective adaptive immune response to vaccination. The synthetic TLR4 agonist glucopyranosyl lipid adjuvant (GLA) synergizes with the squalene oil-in-water emulsion (SE) formulation to induce strong adaptive responses. Although TLR4 signaling through MyD88 and TIR domain-containing adapter inducing IFN-ß are essential for GLA-SE activity, the mechanisms underlying the synergistic activity of GLA and SE are not fully understood. In this article, we demonstrate that the inflammasome activation and the subsequent release of IL-1ß are central effectors of the action of GLA-SE, as infiltration of innate cells into the draining lymph nodes and production of IFN-γ are reduced in ASC-/- animals. Importantly, the early proliferation of Ag-specific CD4+ T cells was completely ablated after immunization in ASC-/- animals. Moreover, numbers of Ag-specific CD4+ T and B cells as well as production of IFN-γ, TNF-α, and IL-2 and Ab titers were considerably reduced in ASC-/-, NLRP3-/-, and IL-1R-/- mice compared with wild-type mice and were completely ablated in TLR4-/- animals. Also, extracellular ATP, a known trigger of the inflammasome, augments Ag-specific CD4+ T cell responses, as hydrolyzing it with apyrase diminished adaptive responses induced by GLA-SE. These data thus demonstrate that GLA-SE adjuvanticity acts through TLR4 signaling and NLRP3 inflammasome activation to promote robust Th1 and B cell responses to vaccine Ags. The findings suggest that engagement of both TLR and inflammasome activators may be a general paradigm for induction of robust CD4 T cell immunity with combination adjuvants such as GLA-SE.


Assuntos
Adjuvantes Imunológicos/farmacologia , Antígenos/imunologia , Linfócitos B/imunologia , Inflamassomos/imunologia , Células Th1/imunologia , Receptor 4 Toll-Like/imunologia , Vacinas/imunologia , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Proteínas Adaptadoras de Sinalização CARD/genética , Feminino , Glucosídeos/imunologia , Imunidade Humoral , Interferon beta/imunologia , Interferon gama/imunologia , Interleucina-1beta/metabolismo , Interleucina-2/imunologia , Lipídeo A/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Receptores Tipo I de Interleucina-1/genética , Esqualeno/imunologia , Receptor 4 Toll-Like/agonistas , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/imunologia , Vacinação
20.
J Immunol Res ; 2018: 3487095, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29682589

RESUMO

Norovirus (NoV) is a main cause of acute gastroenteritis across all ages worldwide. NoV vaccine candidates currently in clinical trials are based on noninfectious highly immunogenic virus-like particles (VLPs) delivered intramuscularly (IM). Since NoV is an enteric pathogen, it is likely that mucosal immunity has a significant role in protection from infection in the intestine. Due to the fact that IM delivery of NoV VLPs does not generate mucosal immunity, we investigated whether NoV genotype GII.4 VLPs coadministered with aluminum hydroxide (Al(OH)3) or monophosphoryl lipid A (MPLA) would induce mucosal antibodies in mice. Systemic as well as mucosal IgG and IgA antibodies in serum and intestinal and nasal secretions were measured. As expected, strong serum IgG, IgG1, and IgG2a antibodies as well as a dose sparing effect were induced by both Al(OH)3 and MPLA, but no mucosal IgA antibodies were detected. In contrast, IN immunization with GII.4 VLPs without an adjuvant induced systemic as well as mucosal IgA antibody response. These results indicate that mucosal delivery of NoV VLPs is needed for induction of mucosal responses.


Assuntos
Infecções por Caliciviridae/imunologia , Norovirus/fisiologia , Vacinas Virais/imunologia , Adjuvantes Imunológicos , Hidróxido de Alumínio/imunologia , Animais , Anticorpos Antivirais/metabolismo , Infecções por Caliciviridae/prevenção & controle , Feminino , Humanos , Imunidade Humoral , Imunidade nas Mucosas , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Lipídeo A/análogos & derivados , Lipídeo A/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Vacinação , Vacinas de Partículas Semelhantes a Vírus
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