A Perspective on the Roles of Adjuvants in Developing Highly Potent COVID-19 Vaccines.

Several countries have made unremitting efforts to develop an optimal vaccine in the fight against coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). With the increasing occurrence of SARS-CoV-2 variants, current vaccines show decreased neutralizing activities, especially towards the Omicron variant. In this context, adding appropriate adjuvants to COVID-19 vaccines can substantially reduce the number of required doses and improve efficacy or cross-neutralizing protection. We mainly focus on research progress and achievements associated with adjuvanted COVID-19 subunit and inactivated vaccines. We further compare the advantages and disadvantages of different adjuvant formulations in order to provide a scientific reference for designing an effective strategy for future vaccine development.

Polyethylenimine-coated PLGA nanoparticles-encapsulated Angelica sinensis polysaccharide as an adjuvant for H9N2 vaccine to improve immune responses in chickens compared to Alum and oil-based adjuvants.

Inactivated H9N2 influenza vaccines required adjuvants to induce strong immune responses to protect poultry from the infections of H9N2 influenza viruses. Recently, positively charged nanoparticles-based adjuvant delivery systems have been extensively investigated as the novel vaccine adjuvant due to the protection antigens and drugs from degradation, promoting antigens and drugs uptake by antigen presenting cells (APCs), and inducing strong humoral and cellular immune responses. In this study, the immunostimulant Angelica sinensis polysaccharide (ASP) was encapsulated into Poly (lactic-co-glycolic acid) PLGA nanoparticles, and the Polyethylenimine (PEI) was coated on the nanoparticles to develop a novel adjuvant (ASP-PLGA-PEI). To further investigate the adjuvant activities of ASP-PLGA-PEI nanoparticles for H9N2 vaccines in chickens and compare the adjuvant activities of nanoparticles adjuvant and conventional adjuvants (Alum and oil-based adjuvant), the H9N2 antigen was incubated with three different adjuvants and then immunized with chickens to evaluate the ability of inducing humoral and cellular immune responses. The results revealed that compared to Alum adjuvant, ASP-PLGA-PEI nanoparticles adjuvant stimulated higher antibody responses, promoted the activation of CD4+ T cells and CD8+ T cells, increased the expression of Th1 cytokines IFN-γ. Compared to oil-based adjuvant (ISA-206), ASP-PLGA-PEI nanoparticles adjuvant induced comparable antibody immune responses at later period after immunization, improved the activation of CD4+ T cells and CD8+ T cells. Therefore, compared to Alum and oil-based adjuvant, the ASP-PLGA-PEI nanoparticles serve as an efficient adjuvant for H9N2 vaccine and have the potential to induce vigorous humoral and cellular immune responses in chickens.

Campylobacter-derived ligands induce cytokine and chemokine expression in chicken macrophages and cecal tonsil mononuclear cells.

Campylobacter jejuni colonizes the chicken gut at a high density without causing disease. However, consumption of poultry products contaminated with this bacterium causes gastroenteritis in humans. Therefore, it is critically important to reduce the Campylobacter burden in poultry products to prevent transmission to humans. Evidence indicates that enhancing intestinal mucosal immune responses is of paramount importance for preventing or reducing Campylobacter colonization in chickens. In view of this, the present study was undertaken to evaluate host responses to different C. jejuni-derived ligands, including lipooligosaccharide (LOS), outer membrane proteins (OMPs), and genomic DNA, with the ultimate goal of identifying a ligand with potent immunostimulatory capacity to serve as a mucosal vaccine adjuvant against enteric infections in chickens. The results revealed that C. jejuni pathogen-associated molecular patterns (PAMPs) varied in their ability to induce the expression of cytokines and chemokines in chicken macrophages and cecal tonsil mononuclear cells and nitric oxide production in macrophages. In addition, C. jejuni OMPs demonstrated superior activity over LOS and DNA ligands in eliciting cytokine expression associated with T helper (Th)1 and Th2 responses (interferon [IFN]-γ and interleukin [IL]-13, respectively), in addition to expression of pro-inflammatory cytokines (IL-1ß), chemokine (CXCLi2), and regulatory cytokines (IL-10 and TGFß1/4) in cecal tonsil cells. Importantly, in addition to their ability to induce innate responses, OMPs could also function as antigens to elicit C. jejuni-specific antibody responses and thereby confer dual protection against C. jejuni infection. Further studies are required to assess the protective efficacy of C. jejuni OMPs against C. jejuni infection in chickens.

[Aluminium adjuvant exposure through vaccines in France in 2018]. / Exposition à l'aluminium vaccinal en France en 2018.

OBJECTIVES: Aluminum-containing vaccine adjuvants stimulate an adequate immune response to vaccination. The safety and rapid elimination of these molecules, a guarantee of their safe use for several decades, have been challenged by a growing number of studies over the last 20 years. Evaluation of exposure to aluminum adjuvants of an individual is thus essential. The current review answers the following questions: what is the exposure of aluminum adjuvants of an individual vaccinated in France? What are the factors of variation? METHODS: To evaluate the immunization exposure to aluminum for a vaccinee in France, we used the 2018 vaccination schedule and the Social Security database for vaccines reimbursed that year. French mandatory and recommended vaccines for an individual who does not travel abroad and has no particular professional obligations have been taken into account. RESULTS: Our results show that an individual following the vaccination requirements and recommendations of 2018 receives between 2545 and 7735µg of Al3+ during his lifetime, and at least 50% before the age of 1year. Exposure varies with age, weight, sex, and choice of administered vaccines. CONCLUSION: Vaccines with higher doses of aluminum are mainly injected at the beginning of life. Women receive a proportionately larger dose than men. The most reimbursed vaccines are often those with the highest amount of aluminum salts.

27Al and 31P NMR spectroscopy method development to quantify aluminum phosphate in adjuvanted vaccine formulations.

A novel qNMR method is described for the quantitative determination of total aluminum and phosphate in aluminum phosphate (AlPO4) adjuvanted vaccine samples using solution 27Al and 31P nuclear magnetic resonance (NMR) spectroscopy. External standard calibrations of AlPO4 solutions established excellent linearity in the range of 15-40 × 10-3 M and additional studies determined the level of detection for both nuclei. A commercialized combination vaccine product (Quadracel®), along with several individual adsorbed antigen components used in the vaccine were employed as model systems for method development. The developed method is also capable of quantitating the free phosphate (i.e. the fraction not bound to AlPO4 particles) in adjuvanted vaccines. This study is the first demonstration of a solution NMR method that is suitable for measuring total aluminum, and free and total phosphate concentrations in vaccine formulations consisting of antigen(s) adsorbed to aluminum adjuvant, in a single analytical workflow.

Effect of 2 Emulsion-Based Adjuvants on the Structure and Thermal Stability of Staphylococcus aureus Alpha-Toxin.

The effects of 2 squalene-based emulsion adjuvant systems (MedImmune emulsion 0 [ME.0] and Stable Emulsion [SE]) on the structure and stability of the recombinant protein antigen alpha-toxin (AT), a potential vaccine candidate for Staphylococcus aureus infection, were investigated using Fourier-transform infrared spectroscopy and both steady-state and time-resolved intrinsic fluorescence spectroscopy as well as differential scanning calorimetry (DSC). A component study, performed to identify the effects of the individual emulsion's components, showed negligible interactions between AT and ME.0. DSC analysis showed the ME.0 emulsion thermally destabilized AT, probably because of changes in the buffer composition of AT upon mixing. The SE emulsion caused increased alpha-helix and decreased beta-sheet content in AT, and a significant blue shift in the fluorescence spectra relative to that of AT in solution. DSC analysis showed SE exerted a dramatic thermal stabilization effect on AT, probably attributable to an interaction between AT and SE. Size exclusion chromatography showed a complete loss in the recovery of AT when mixed with SE, but not ME.0, indicating a high degree of interaction with SE. This work successfully characterized the biophysical properties of AT in the presence of 2 emulsion adjuvants including a component study to rationalize how emulsion components affect protein antigen stability.

The antibody response in the bovine mammary gland is influenced by the adjuvant and the site of subcutaneous vaccination.

Intramammary infections in cattle resulting in mastitis have detrimental effects on cows' well-being, lifespan and milk production. In the host defense against S. aureus mastitis antibodies are thought to play an important role. To explore potential ways to increase antibody titers in the bovine mammary gland the effects of various adjuvants on the magnitude, isotype, and neutralizing capacity of antibodies produced following subcutaneous vaccine administration at different immunization sites were analyzed. In this study, α-toxoid was used as a model antigen and formulated in three different alum-based adjuvants: Alum-Saponin, Alum-Oil, and Alum-Saponin-Oil. Vaccines were administered near the suspensory ligament of the udder or in the lateral triangular area of the neck. At both immunization sites, immunization with α-toxoid in Alum-Saponin-Oil resulted in higher specific antibody titers in milk and serum as compared with Alum-Oil and Alum-Saponin, without favoring an IgG1, IgG2, or IgA response. Furthermore, the neutralizing capacity of milk serum and serum following immunization near the udder and in the neck was higher when Alum-Saponin-Oil was used as adjuvant compared with Alum-Oil and Alum-Saponin. Prime immunizations near the udder effectively increased both antibody isotype titers and neutralization titers, while prime plus boost immunizations were required to induce similar effects following immunization in the neck. Results indicate that subcutaneous administration of an Alum-Saponin-Oil based vaccine near the udder could be further explored for the development of a one-shot vaccination strategy to efficiently increase intramammary antibody responses.

Down selecting adjuvanted vaccine formulations: a comparative method for harmonized evaluation.

BACKGROUND: The need for rapid and accurate comparison of panels of adjuvanted vaccine formulations and subsequent rational down selection, presents several challenges for modern vaccine development. Here we describe a method which may enable vaccine and adjuvant developers to compare antigen/adjuvant combinations in a harmonized fashion. Three reference antigens: Plasmodium falciparum apical membrane antigen 1 (AMA1), hepatitis B virus surface antigen (HBsAg), and Mycobacterium tuberculosis antigen 85A (Ag85A), were selected as model antigens and were each formulated with three adjuvants: aluminium oxyhydroxide, squalene-in-water emulsion, and a liposome formulation mixed with the purified saponin fraction QS21. RESULTS: The nine antigen/adjuvant formulations were assessed for stability and immunogenicity in mice in order to provide benchmarks against which other formulations could be compared, in order to assist subsequent down selection of adjuvanted vaccines. Furthermore, mouse cellular immune responses were analyzed by measuring IFN-γ and IL-5 production in splenocytes by ELISPOT, and humoral responses were determined by antigen-specific ELISA, where levels of total IgG, IgG1, IgG2b and IgG2c in serum samples were determined. CONCLUSIONS: The reference antigens and adjuvants described in this study, which span a spectrum of immune responses, are of potential use as tools to act as points of reference in vaccine development studies. The harmonized methodology described herein may be used as a tool for adjuvant/antigen comparison studies.

Preformulation studies with the Escherichia coli double mutant heat-labile toxin adjuvant for use in an oral vaccine.

Double mutant heat-labile toxin (dmLT) is a promising adjuvant for oral vaccine administration. The aims of our study were to develop sensitive methods to detect low concentrations of dmLT and to use the assays in preformulation studies to determine whether dmLT remains stable under conditions encountered by an oral vaccine. We developed a sandwich ELISA specific for intact dmLT and a sensitive SDS-PAGE densitometry method, and tested stability of dmLT in glass and plastic containers, in saliva, at the pH of stomach fluid, and in high-osmolarity buffers. The developed ELISA has a quantification range of 62.5 to 0.9ng/mL and lower limit of detection of 0.3ng/mL; the limit of quantification of the SDS-PAGE is 10µg/mL. This work demonstrates the application of dmLT assays in preformulation studies to development of an oral vaccine containing dmLT. Assays reported here will facilitate the understanding and use of dmLT as an adjuvant.

Enhanced Release of Immunostimulating ß-1,3- Glucan by Autodigestion of the Lingzhi Medicinal Mushroom, Ganoderma lingzhi (Agaricomycetes).

Ganoderma lingzhi is a widely used medicinal mushroom that has antioxidative effects, ameliorates insulin resistance, and improves quality of life in patients with metabolic syndrome. Potentiation of immunity is also a major function of G. lingzhi, and this has been applied in patients with cancer. Supplementing G. lingzhi into foods reduced the metastasis of cancer cells. ß-l,3-glucan is an important bioactive component of G. lingzhi. In this study we enhanced the solubilization ofimmunostimulating ß-l,3-glucan by autodigestion of G. lingzhi. Fruiting bodies of G. lingzhi were disrupted and suspended in distilled water, then autodigested at 37°C for 24 hours. The resulting suspension was dried by spray drying. To assess the solubilization of ß-l,3-glucan by autodigestion, cold and hot water extracts and sodium hydroxide extracts of G. lingzhi were prepared with and without autodigestion. Sodium hydroxide extracts were neutralized and dialyzed against distilled water. The resulting soluble and precipitated fractions were collected. Chemical, biochemical, and immunochemical characteristics of the extracts were compared. The yields of cold water extracts of autodigested and native G. lingzhi were significantly lower than the other extracts. Glucose was the major sugar component of the hot water extract, cold alkali extract (CAS), and the cold hydroxide extract insoluble in neutral aqueous condition (CASP) of the autodigested and native G. lingzhi. Nuclear magnetic resonance analysis revealed branched ß-glucans in the hot water extract and CAS of the autodigested and native G. lingzhi. By contrast, the CASP of the autodigested and native G. lingzhi comprised mainly mixtures of linear α-l,3-glucans and linear ß-l,3-glucans. Immunostimulation by ß-l,3-glucan was examined by limulus factor G activation, dectin-1 binding, and anti-ß-glucan antibody binding. Comparing relative activity, immunostimulating ß-l,3-glucan was detected in the hot water extract, rather than the CAS, of autodigested and native G. lingzhi. Immunostimulating of ß-glucan was also detected in the cold water extract of the autodigested G. lingzhi. These findings demonstrate that autodigestion is a useful processing protocol for enhancing the usefulness of G. lingzhi as a functional food.

Quantitation of Monophosphorylated Lipid A in the Oil-in-Water Adjuvant Delivery Systems Using Transesterification and GC-MS.

Vaccine delivery systems play pivotal role in effective antigen delivery. These systems often contain adjuvants that stimulate specific immune response and are important for vaccines' efficacy and safety. Oil-in-water vaccine delivery lipid emulsion systems containing monophosphoryl lipid A (MPLA) as immune modulator have been extensively investigated in vaccine trials. Herein, we describe a simple orthogonal method, for quantitative measurement of MPLA in an oil-in-water lipid delivery system using direct transesterification reaction followed by gas-chromatography-mass spectrometry analysis. In this protocol, the transesterification reaction results in the release of fatty acid methyl esters followed by gas-chromatography-mass spectrometry-based targeted quantification of the specific 3-hydroxytetradecanoate fatty acid methyl ester to measure the concentration of MPLA in an oil-in-water lipid emulsion system.

A simple fluorescence-based assay for quantification of the Toll-Like Receptor agonist E6020 in vaccine formulations.

Despite the generally accepted immunostimulatory effect of Toll-Like Receptor 4 (TLR4) agonists and their value as vaccine adjuvants, there remains a demand for fast and easy quantification assays for these TLR4 agonists in order to accelerate and improve vaccine formulation studies. A new medium-throughput method was developed for the quantification of the TLR4 agonist, E6020, independent of the formulation composition. The assay uses a fluorescent hydrazide (DCCH) to label the synthetic lipopolysaccharide (LPS) analog E6020 through its diketone groups. This novel, low-cost, and fluorescence based assay may obviate the need for traditional approaches that primarily rely on Fourier transform infrared spectroscopy (FTIR) or mass spectrometry. The experiments were performed in a wide diversity of vaccine formulations containing E6020 to assess method robustness and accuracy. The assay was also expanded to evaluate the loading efficiency of E6020 in poly(lactic-co-glycolic acid) (PLGA) micro-particles.

p-sulfonated calix[8]arene functionalized graphene as a "turn on" fluorescent sensing platform for aconitine determination.

This work reports a novel method for the determination of aconitine through the competitive host-guest interaction between p-sulfonated calix[8]arene (SCX8) and signal probe/target molecules by using SCX8 functionalized reduced graphene oxide (SCX8-RGO) as a receptor. Three dyes (ST, RhB, BRB) and aconitine were selected as the probe and target molecules, respectively. The formation of SCX8-RGO·ST, SCX8-RGO·RhB, and SCX8-RGO·BRB complexes greatly decreases the fluorescence emission of ST, RhB, and BRB. The aconitine/SCX8 complex possesses a higher binding constant than ST/SCX8, RhB/SCX8, and BRB/SCX8 complexes, thus the dye in the SCX8 cavity can be replaced by aconitine to revert the fluorescence emission of SCX8-RGO·dye, leading to a "switch-on" fluorescence response. The fluorescence intensity of SCX8-RGO·ST, SCX8-RGO·RhB, and SCX8-RGO·BRB complexes increased linearly with increasing concentration of aconitine ranging from 1.0 to 14.0µM, 2.0-16.0µM, and 1.0-16.0µM, respectively. Based on the competitive host-guest interaction, the proposed detection method for aconitine showed detection limits of 0.28µM, 0.60µM, and 0.37µM, respectively, and was successfully applied for the determination of aconitine in human serum samples with good recoveries from 95.1% to 104.8%. The proposed method showed high selectivity for aconitine beyond competitive binding analytes. In addition, the inclusion complex of the SCX8/aconitine was studied by the molecular docking and molecular dynamics simulation, which indicated that the phenyl ester group of the aconitine molecule was included into the SCX8 cavity.

Proteome profiling of the growth phases of Leishmania pifanoi promastigotes in axenic culture reveals differential abundance of immunostimulatory proteins.

Leishmaniasis is a term that encompasses a compendium of neglected tropical diseases caused by dimorphic and digenetic protozoan parasites from the genus Leishmania (Kinetoplastida: Trypanosomatidae). The clinical manifestations of neotropical cutaneous leishmaniasis (NCL) caused by Leishmania pifanoi and other species of the "Leishmania mexicana complex" mainly correspond to anergic diffuse cutaneous leishmaniasis (ADCL), which is the origin of considerable morbidity. Despite the outstanding advances in the characterization of the trypanosomatid genomes and proteomes, the biology of this species has been scarcely explored. However, the close relation of L. pifanoi to the sequenced species L. mexicana and others included in the "L. mexicana complex" allowed us to perform a two-dimension electrophoresis (2DE) approach to the promastigote proteome at the differential expression level. Protein identifications were performed by matrix-assisted laser desorption-ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF). This insight has revealed similarities and differences between L. pifanoi and other species responsible for cutaneous and visceral leishmaniasis. Interestingly, certain proteins that were previously described as immunostimulatory (elongation factor 1ß, trypanothione peroxidase, heat shock protein 70, enolase, GDP-forming succinyl-CoA and aldehyde dehydrogenase) are more abundant in the final growth stages of promastigotes (late-logarithmic and/or stationary phase) in the case of L. pifanoi.

Identification of QS-21 as an Inflammasome-activating Molecular Component of Saponin Adjuvants.

Many immunostimulants act as vaccine adjuvants via activation of the innate immune system, although in many cases it is unclear which specific molecules contribute to the stimulatory activity. QS-21 is a defined, highly purified, and soluble saponin adjuvant currently used in licensed and exploratory vaccines, including vaccines against malaria, cancer, and HIV-1. However, little is known about the mechanisms of cellular activation induced by QS-21. We observed QS-21 to elicit caspase-1-dependent IL-1ß and IL-18 release in antigen-presenting cells such as macrophages and dendritic cells when co-stimulated with the TLR4-agonist adjuvant monophosphoryl lipid A. Furthermore, our data suggest that the ASC-NLRP3 inflammasome is responsible for QS-21-induced IL-1ß/IL-18 release. At higher concentrations, QS-21 induced macrophage and dendritic cell death in a caspase-1-, ASC-, and NLRP3-independent manner, whereas the presence of cholesterol rescued cell viability. A nanoparticulate adjuvant that contains QS-21 as part of a heterogeneous mixture of saponins also induced IL-1ß in an NLRP3-dependent manner. Interestingly, despite the role NLRP3 plays for cellular activation in vitro, NLRP3-deficient mice immunized with HIV-1 gp120 and QS-21 showed significantly higher levels of Th1 and Th2 antigen-specific T cell responses and increased IgG1 and IgG2c compared with wild type controls. Thus, we have identified QS-21 as a nonparticulate single molecular saponin that activates the NLRP3 inflammasome, but this signaling pathway may contribute to decreased antigen-specific responses in vivo.

Biological activities and pharmacokinetics of aconitine, benzoylaconine, and aconine after oral administration in rats.

Aconitine (AC), benzoylaconine (BAC), and aconine (ACN) are three representative alkaloids in Aconitum tubers. Knowing that the drug disposal process in vivo is closely related to the toxicity and efficacy of a drug, it is important to classify the disposal properties of these alkaloids. In this study, the pharmacokinetics of the three alkaloids was investigated. The results showed that the three alkaloids could be quickly absorbed, especially BAC, whose Tmax was 0.31 ± 0.17 h. Their Cmax was 10.99, 3.99, and 4.29 ng·mL(-1) respectively, indicating that AC had better absorption than BAC and ACN. Subsequently, we further investigated their absorption mechanism using the Caco-2 cell monolayer model in vitro. The results showed that they were poorly absorbed, and the absorption of AC and BAC was inhibited by P-gp, while the absorption of ACN was in a form of passive diffusion. The t1/2 of AC, BAC and ACN was 1.41, 9.49, and 3.32 h, respectively, indicating that the metabolic or excretion rate of AC was quicker than that of BAC and ACN. Therefore, their metabolic stability was further investigated by using rat liver microsomes in vitro, which showed that AC was easier to be metabolized than BAC and ACN. The excretion experiments showed that AC and ACN were primarily excreted in urine, while BAC was excreted in faeces. In addition, the results of tissue distribution experiments showed that the three alkaloids distributed throughout all the organs, although the distribution rate of AC was slower than that of BAC and ACN. Copyright © 2015 John Wiley & Sons, Ltd.

Plantas medicinais como alternativa terapêutica para aumento da resistência imunológica / Medicinal plants as an alternative therapy to increase immune resistance

The immune system modulation in disease prevention and treatment has been the subject of interest for many years, so that many studies have emphasized the immunomodulatory drugs research, especially from natural origin. So, in order to provide a current information source, this paper presents a review of the medicinal plants with immunostimulatory activity evidenced in recent studies. It was found in these studies that a wide variety of families presents species with immunostimulating activity, however the Euphorbiaceae, Fabaceae Moraceae, Rubiaceae and Zingiberaceae families were more reported.(AU)

A modulação do sistema imune na prevenção e tratamento de doenças têm sido um assunto de interesse há muitos anos, de modo que muitas pesquisas tem dado ênfase à investigação de novas drogas imunomodularoras, sobretudo de origem natural. Portanto, a fim de proporcionar uma fonte de informação atualizada, o presente artigo apresenta uma revisão de plantas medicinais com atividade imunoestimulante evidenciadas em estudos recentes. Verificou-se nestes estudos que uma grande variedade de famílias apresentam espécies com atividade imunoestimulante, contudo, as famílias Euphorbiaceae, Fabaceae Moraceae, Rubiaceae and Zingiberaceae foram as mais relatadas.(AU)

Quantitation of the immunological adjuvants, monophosphoryl lipid A and Quil A in poly (lactic-co-glycolic acid) nanoparticles using high performance liquid chromatography with evaporative light scattering detection.

Monophosphoryl lipid A (MPL) and Quil A are two immunological adjuvants commonly used in vaccines. At present no simple, validated methods for the quantification of Quil A and MPL have been previously reported therefore the aim of the current study was to develop a simple, fast and validated method to quantify MPL and Quil A using high performance liquid chromatography evaporative light scattering detection (HPLC-ELSD). The HPLC-ELSD technique was carried out using a ZORBAX Eclipse XDB-C8 column (2.1×50 mm; particle size, 3.5 µm) in an isocratic elution mode at 25 °C. MPL was eluted at a retention time of 1.8 min with methanol-water as the mobile phase and a detector temperature of 75 °C. Quil A was resolved as three peaks with retention times of 4.1, 5.5 and 6.4 min with a detector temperature of 30 °C and with water-acetonitrile and 0.01% formic acid as the mobile phase. The nebulizer pressure and gain were set at 3.5 bar and 10, respectively. Calibration curves plotted for both the adjuvants had an R(2)>0.997. Accuracy, intra- and inter-day precision were within the accepted limits. The limit of detection for MPL and Quil A were calculated as 1.343 and 2.06 µg/mL, respectively. The limit of quantification was 2.445 for MPL and 8.97 µg/mL for Quil A. This analytical method was used to quantify the entrapment and in vitro release of MPL and Quil A in a poly lactic-co-glycolic acid (PLGA) nanoparticle vaccine.

Caracterização fenotípica e funcional da resposta imune de voluntários imunizados contra influenza A (H1N1)pdm09

A primeira pandemia de gripe do século XXI foi declarada em junho de 2009 pela Organização Mundial de Saúde (OMS)e representou uma grande ameaça à saúde da população mundial. E uma vez que as primeiras vacinas foram licenciadas, tornou-se iminente a busca de informações mais aprofundadas sobre os mecanismos imunológicos desencadeados por estas vacinas, considerando ainda a presença ou ausência de adjuvante em sua formulação. Neste contexto,o presente trabalho objetiva avaliar aspectos fenotípicos e funcionais das imunidades celular e humoral em indivíduos vacinados contra o vírus da Influenza A (H1N1)pdm09. Para isso, 20 voluntários saudáveis foram imunizados contra H1N1pandêmico, na presença (n=10) ou ausência (n=10) de adjuvante (óleo em água), e avaliados nos tempos 0, 1, 3, 7, 30 dias pós-imunização. Desta forma, diversos parâmetros foram utilizados para caracterizar a imunidade desenvolvida por estas vacinas, dentre os quais, avaliação clínica e hematológica, sorologia, imunofenotipagem celular e análise de citocinas intracelulares e plasmáticas.

Coletivamente, os resultados demonstraram um perfil imunológico bastante distinto desencadeado pelas vacinas, embora ambas tenham induzido níveis semelhantes de anticorpos e persistência destes 6 meses após a vacinação.A vacina com adjuvante foi capaz de induzir uma resposta mais intensa de células da imunidade inata, com significativa ativação da imunidade celular, principalmente de linfócitos T CD4+e aumento na frequência de linfócitos B. Além disso, esta vacina induziu uma expressiva produção de citocinas, apresentando um padrão misto de resposta Th1/ Th2 neste grupo. Por outro lado, a vacina sem adjuvante, desencadeou uma resposta mais discreta e tardia, com menor ativação da imunidade inata e fraca indução de linfócitos T, porém com elevada ativação de linfócitos B. Esta vacina também causou uma menor produção de citocinas, desencadeando um perfil pouco inflamatório neste grupo. Em suma, as vacinas induziram perfis distintos de respostas imune inata e adaptativa, o que parece estar relacionado à presença ou ausência de adjuvante. Contudo, é importante enfatizar que ambas as vacinas foram capazes de induzir uma resposta eficaz no que diz respeito à imunogenicidade, o que sugerei munidade protetora contra o vírus H1N1pandêmico.

Caracterização fenotípica e funcional da resposta imune de voluntários imunizados contra influenza A (H1N1)pdm09 / Phenotypic and functional characterization of the immune response volunteers immunized against influenza A (H1N1) pdm09

A primeira pandemia de gripe do século XXI foi declarada em junho de 2009 pela Organização Mundial de Saúde (OMS)e representou uma grande ameaça à saúde da população mundial. E uma vez que as primeiras vacinas foram licenciadas, tornou-se iminente a busca de informações mais aprofundadas sobre os mecanismos imunológicos desencadeados por estas vacinas, considerando ainda a presença ou ausência de adjuvante em sua formulação. Neste contexto,o presente trabalho objetiva avaliar aspectos fenotípicos e funcionais das imunidades celular e humoral em indivíduos vacinados contra o vírus da Influenza A (H1N1)pdm09. Para isso, 20 voluntários saudáveis foram imunizados contra H1N1pandêmico, na presença (n=10) ou ausência (n=10) de adjuvante (óleo em água), e avaliados nos tempos 0, 1, 3, 7, 30 dias pós-imunização. Desta forma, diversos parâmetros foram utilizados para caracterizar a imunidade desenvolvida por estas vacinas, dentre os quais, avaliação clínica e hematológica, sorologia, imunofenotipagem celular e análise de citocinas intracelulares e plasmáticas...

Coletivamente, os resultados demonstraram um perfil imunológico bastante distinto desencadeado pelas vacinas, embora ambas tenham induzido níveis semelhantes de anticorpos e persistência destes 6 meses após a vacinação.A vacina com adjuvante foi capaz de induzir uma resposta mais intensa de células da imunidade inata, com significativa ativação da imunidade celular, principalmente de linfócitos T CD4+e aumento na frequência de linfócitos B. Além disso, esta vacina induziu uma expressiva produção de citocinas, apresentando um padrão misto de resposta Th1/ Th2 neste grupo. Por outro lado, a vacina sem adjuvante, desencadeou uma resposta mais discreta e tardia, com menor ativação da imunidade inata e fraca indução de linfócitos T, porém com elevada ativação de linfócitos B. Esta vacina também causou uma menor produção de citocinas, desencadeando um perfil pouco inflamatório neste grupo. Em suma, as vacinas induziram perfis distintos de respostas imune inata e adaptativa, o que parece estar relacionado à presença ou ausência de adjuvante. Contudo, é importante enfatizar que ambas as vacinas foram capazes de induzir uma resposta eficaz no que diz respeito à imunogenicidade, o que sugerei munidade protetora contra o vírus H1N1pandêmico...