RESUMO
Multivalent presentation of ligands often enhances receptor activation and downstream signalling. DNA origami offers a precise nanoscale spacing of ligands, a potentially useful feature for therapeutic nanoparticles. Here we use a square-block DNA origami platform to explore the importance of the spacing of CpG oligonucleotides. CpG engages Toll-like receptors and therefore acts to activate dendritic cells. Through in vitro cell culture studies and in vivo tumour treatment models, we demonstrate that square blocks induce Th1 immune polarization when CpG is spaced at 3.5 nm. We observe that this DNA origami vaccine enhances DC activation, antigen cross-presentation, CD8 T-cell activation, Th1-polarized CD4 activation and natural-killer-cell activation. The vaccine also effectively synergizes with anti-PD-L1 for improved cancer immunotherapy in melanoma and lymphoma models and induces long-term T-cell memory. Our results suggest that DNA origami may serve as a platform for controlling adjuvant spacing and co-delivering antigens in vaccines.
Assuntos
Vacinas Anticâncer , Oligodesoxirribonucleotídeos , Animais , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , Camundongos , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/farmacologia , DNA/química , DNA/imunologia , Células Dendríticas/imunologia , Humanos , Camundongos Endogâmicos C57BL , Ilhas de CpG , Vacinas de DNA/química , Vacinas de DNA/imunologia , Vacinas de DNA/farmacologia , Linfócitos T CD8-Positivos/imunologia , Vacinação/métodos , Linhagem Celular Tumoral , FemininoRESUMO
BACKGROUND: Canine distemper virus (CDV), which is highly infectious, has caused outbreaks of varying scales in domestic and wild animals worldwide, so the development of a high-efficiency vaccine has broad application prospects. Currently, the commercial vaccine of CDV is an attenuated vaccine, which has the disadvantages of a complex preparation process, high cost and safety risk. It is necessary to develop a safe and effective CDV vaccine that is easy to produce on a large scale. In this study, sequences of CDV haemagglutinin (HA) from the Yanaka strain were aligned, and three potential linear sequences, termed YaH3, YaH4, and YaH5, were collected. To increase the immunogenicity of the epitopes, ferritin was employed as a self-assembling nanoparticle element. The ferritin-coupled forms were termed YaH3F, YaH4F, and YaH5F, respectively. A full-length HA sequence coupled with ferritin was also constructed as a DNA vaccine to compare the immunogenicity of nanoparticles in prokaryotic expression. RESULT: The self-assembly morphology of the proteins from prokaryotic expression was verified by transmission electron microscopy. All the proteins self-assembled into nanoparticles. The expression of the DNA vaccine YaHF in HEK-293T cells was also confirmed in vitro. After subcutaneous injection of epitope nanoparticles or intramuscular injection of DNA YaHF, all vaccines induced strong serum titres, and long-term potency of antibodies in serum could be detected after 84 days. Strong anti-CDV neutralizing activities were observed in both the YaH4F group and YaHF group. According to antibody typing and cytokine detection, YaH4F can induce both Th1 and Th2 immune responses. The results of flow cytometry detection indicated that compared with the control group, all the immunogens elicited an increase in CD3. Simultaneously, the serum antibodies induced by YaH4F and YaHF could significantly enhance the ADCC effect compared with the control group, indicating that the antibodies in the serum effectively recognized the antigens on the cell surface and induced NK cells to kill infected cells directly. CONCLUSIONS: YaH4F self-assembling nanoparticle obtained by prokaryotic expression has no less of an immune effect than YaHF, and H4 has great potential to become a key target for the easy and rapid preparation of epitope vaccines.
Assuntos
Vírus da Cinomose Canina , Ferritinas/química , Hemaglutininas Virais , Nanopartículas Metálicas/química , Vacinas de DNA , Animais , Anticorpos Antivirais/química , Anticorpos Antivirais/imunologia , Chlorocebus aethiops , Citocinas/metabolismo , Cinomose/prevenção & controle , Vírus da Cinomose Canina/química , Vírus da Cinomose Canina/imunologia , Cães , Feminino , Células HEK293 , Hemaglutininas Virais/química , Hemaglutininas Virais/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Vacinas de DNA/química , Vacinas de DNA/imunologia , Células VeroRESUMO
Cancer vaccines based on DNA encoding oncogenes have shown great potential in preclinical studies. However, the efficacy of DNA vaccines is limited by their weak immunogenicity because of low cellular internalisation and insufficient activation of dendritic cells (DCs). Calcium phosphate (CP) nanoparticles (NPs) are biodegradable vehicles with low toxicity and high loading capacity of DNA but suffer from stability issues. Here we employed adenosine triphosphate (ATP) as a dual functional agent, i.e. stabiliser for CP and immunological adjuvant, and applied the ATP-modified CP (ACP) NPs to the DNA vaccine. ACP NP-enhanced cellular uptake and improved transfection efficiency of DNA vaccine, and further showed the ability to activate DCs that are critical for them to prime T cells in cancer immunotherapy. As a result, a higher level of antigen-specific antibody with stronger tumour growth inhibition was achieved in mice immunised with the ACP-DNA vaccine. Overall, this one-step synthesised ACP NPs are an efficient nano-delivery system and nano-adjuvant for cancer DNA vaccines.
Assuntos
Trifosfato de Adenosina/química , Adjuvantes Imunológicos/química , Fosfatos de Cálcio/química , Nanopartículas/química , Vacinas de DNA/química , Animais , Reações Antígeno-Anticorpo , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , Linhagem Celular Tumoral , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Imunoterapia , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias/terapia , Transplante Homólogo , Vacinação , Vacinas de DNA/imunologia , Vacinas de DNA/farmacologiaRESUMO
Toxoplasma gondii is an obligate intracellular protozoan parasite that can cause serious public health problems. The development of a safe and effective vaccine against T. gondii is urgently needed to prevent and control the spread of toxoplasmosis. The aim of this study was to evaluate the immune responses induced by a pcGRA14 + pcROP13 vaccine cocktail in BALB/c mice. All groups were immunized intramuscularly three times at two-week intervals. The production of anti-Toxoplasma gondii lysate antigen (TLA) antibodies, lymphocyte proliferation, serum levels of IFN-γ and IL-4 cytokines and the survival time were monitored after vaccination and challenged with the virulent RH strain of T. gondii. The results showed that immunization with the pcGRA14 + pcROP13 DNA vaccine significantly increased the production of specific IgG antibodies and cytokines against toxoplasmosis. Interestingly, high levels of IgG2a and IFN-γ were found in animals vaccinated with DNA vaccine cocktail. Furthermore, immunized mice challenged with the RH strain of T. gondii showed prolonged survival time when compared to control groups (P <0.05). The present study demonstrates the potential of a DNA cocktail vaccine expressing pcGRA14 and pcROP13 in developing specific immune responses and providing effective protection against T. gondii infection.
Assuntos
Adjuvantes Imunológicos/química , Compostos de Alúmen/química , Proteínas de Protozoários/genética , Toxoplasmose/prevenção & controle , Vacinas de DNA/química , Vacinas de DNA/imunologia , Animais , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Protozoários/imunologia , Toxoplasma/genética , Toxoplasma/imunologia , Toxoplasmose/imunologiaRESUMO
In DNA-based therapy research, the conception of a suitable vector to promote the target gene carriage, protection, and delivery to the cell is imperative. Exploring the interactions between polyethylenimine (PEI) and a plasmid DNA can give rise to the formation of suitable complexes for gene release and concomitant protein production. The nanosystems formulation method, based on coprecipitation, seems to be adequate for the conception of nanoparticles with suitable properties (morphology, size, surface charge, and pDNA complexation capacity) for intracellular applications. The developed systems are able of cell uptake, intracellular trafficking, and gene expression, in an extent depending on the ratio of nitrogen to phosphate groups (N/P). It comes that the transfection process can be tailored by this parameter and, therefore, also the therapeutic outcomes. This knowledge contributes for progresses in the development of suitable delivery systems with potential application in DNA vaccines field.
Assuntos
Técnicas de Transferência de Genes , Plasmídeos/administração & dosagem , Plasmídeos/química , Polietilenoimina/química , Vacinas de DNA/administração & dosagem , Cátions , Sistemas de Liberação de Medicamentos , Células HeLa , Humanos , Nanopartículas , Espectroscopia de Infravermelho com Transformada de Fourier , Vacinas de DNA/química , Vacinas de DNA/genéticaRESUMO
Over the last 30â years, genetically engineered DNA has been tested as novel vaccination strategy against various diseases, including human immunodeficiency virus (HIV), hepatitis B, several parasites, and cancers. However, the clinical breakthrough of the technique is confined by the low transfection efficacy and immunogenicity of the employed vaccines. Therefore, carrier materials were designed to prevent the rapid degradation and systemic clearance of DNA in the body. In this context, biopolymers are a particularly promising DNA vaccine carrier platform due to their beneficial biochemical and physical characteristics, including biocompatibility, stability, and low toxicity. This article reviews the applications, fabrication, and modification of biopolymers as carrier medium for genetic vaccines.
Assuntos
Biopolímeros/química , Vacinas de DNA/administração & dosagem , Anticorpos Monoclonais/química , Anticorpos Monoclonais/uso terapêutico , Humanos , Lipossomos/química , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Neoplasias/terapia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Vacinas de DNA/química , Vacinas de DNA/imunologia , Viroses/prevenção & controleRESUMO
The novel corona virus termed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread throughout the globe at a formidable speed, causing tens of millions of cases and more than one million deaths in less than a year of its report in December 2019. Since then, companies and research institutions have raced to develop SARS-CoV-2 vaccines, ranging from conventional viral and protein-based vaccines to those that are more cutting edge, including DNA- and mRNA-based vaccines. Each vaccine exhibits a different potency and duration of efficacy, as determined by the antigen design, adjuvant molecules, vaccine delivery platforms, and immunization method. In this review, we will introduce a few of the leading non-viral vaccines that are under clinical stage development and discuss delivery strategies to improve vaccine efficacy, duration of protection, safety, and mass vaccination.
Assuntos
Vacinas contra COVID-19/administração & dosagem , COVID-19/prevenção & controle , Sistemas de Liberação de Medicamentos/métodos , Vacinas Sintéticas/administração & dosagem , Animais , COVID-19/genética , Vacinas contra COVID-19/química , Vacinas contra COVID-19/genética , Humanos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/química , Vacinas de DNA/genética , Vacinas Sintéticas/química , Vacinas Sintéticas/genéticaRESUMO
Infectious Bronchitis (IB) is an economically important avian disease that considerably threatens the global poultry industry. This is partly, as a result of its negative consequences on egg production, weight gain as well as mortality rate.The disease is caused by a constantly evolving avian infectious bronchitis virus whose isolates are classified into several serotypes and genotypes that demonstrate little or no cross protection. In order to curb the menace of the disease therefore, broad based vaccines are urgently needed. The aim of this study was to develop a recombinant DNA vaccine candidate for improved protection of avian infectious bronchitis in poultry. Using bioinformatics and molecular cloning procedures, sets of monovalent and bivalent DNA vaccine constructs were developed based on the S1 glycoprotein from classical and variants IBV strains namely, M41 and CR88 respectively. The candidate vaccine was then encapsulated with a chitosan and saponin formulated nanoparticle for enhanced immunogenicity and protective capacity. RT-PCR assay and IFAT were used to confirm the transcriptional and translational expression of the encoded proteins respectively, while ELISA and Flow-cytometry were used to evaluate the immunogenicity of the candidate vaccine following immunization of various SPF chicken groups (A-F). Furthermore, histopathological changes and virus shedding were determined by quantitative realtime PCR assay and lesion scoring procedure respectively following challenge of various subgroups with respective wild-type IBV viruses. Results obtained from this study showed that, groups vaccinated with a bivalent DNA vaccine construct (pBudCR88-S1/M41-S1) had a significant increase in anti-IBV antibodies, CD3+ and CD8+ T-cells responses as compared to non-vaccinated groups. Likewise, the bivalent vaccine candidate significantly decreased the oropharyngeal and cloacal virus shedding (p < 0.05) compared to non-vaccinated control. Chickens immunized with the bivalent vaccine also exhibited milder clinical signs as well as low tracheal and kidney lesion scores following virus challenge when compared to control groups. Collectively, the present study demonstrated that bivalent DNA vaccine co-expressing dual S1 glycoprotein induced strong immune responses capable of protecting chickens against infection with both M41 and CR88 IBV strains. Moreso, it was evident that encapsulation of the vaccine with chitosan-saponin nanoparticle further enhanced immune responses and abrogates the need for multiple booster administration of vaccine. Therefore, the bivalent DNA vaccine could serve as efficient and effective alternative strategy for the control of IB in poultry.
Assuntos
Quitosana/imunologia , Infecções por Coronavirus/veterinária , Vírus da Bronquite Infecciosa/imunologia , Doenças das Aves Domésticas/imunologia , Saponinas/imunologia , Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/imunologia , Bronquite/imunologia , Bronquite/prevenção & controle , Bronquite/veterinária , Linfócitos T CD8-Positivos/imunologia , Galinhas , Quitosana/química , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Proteção Cruzada , Imunidade Celular , Imunização Secundária/veterinária , Imunogenicidade da Vacina , Nanopartículas/química , Doenças das Aves Domésticas/prevenção & controle , Saponinas/química , Vacinação/veterinária , Vacinas de DNA/química , Vacinas de DNA/genética , Vacinas Virais/química , Vacinas Virais/genéticaRESUMO
A novel concept in DNA vaccine design is the creation of an inhaled DNA plasmid construct containing a portion of the coronavirus spike protein for treatment and vaccination. The secretion of a spike protein portion will function as a competitive antagonist by interfering with the binding of coronavirus to the angiotensin-converting enzyme 2 (ACE2) receptor. The secreted protein binding to the ACE2 receptor provides a unique mechanism of action for treatment to all strains of coronavirus in naïve patients, by blocking the ACE2 receptor site. An inhaled plasmid DNA vaccine replicates the route of lung infection taken by coronavirus with transfected cells secreting spike protein portions to induce immunity. Unlike most DNA vaccines with intracellular antigen presentation through MHC I, the current vaccine relies on the secreted proteins presentation through MHC II as well as MHC I to induce immunity. Lung specific production of vaccine particles by inhaled plasmid DNA is appealing since it may have limited systemic side effects, and may induce both humoral and cytotoxic immunity. Finally, the ease and ability to rapidly produce this plasmid construct makes this an ideal solution for managing the emerging threat of coronavirus.
Assuntos
Glicoproteína da Espícula de Coronavírus/imunologia , Vacinas de DNA/administração & dosagem , Vacinas de DNA/uso terapêutico , Vacinas Virais/administração & dosagem , Vacinas Virais/uso terapêutico , Administração Intranasal , Betacoronavirus/genética , Betacoronavirus/imunologia , COVID-19 , Vacinas contra COVID-19 , Quitosana , Infecções por Coronavirus/prevenção & controle , Humanos , Pandemias , Pneumonia Viral , SARS-CoV-2 , Vacinação/métodos , Vacinas de DNA/químicaRESUMO
AIM: To estimate immunogenicity and antitumor effect of new DNA vaccine against neuroblastoma using tyrosine hydroxylase as an antigen and linear polyethylenimine (PEI) 20 kDa as a synthetic DNA carrier in syngeneic mouse tumor model. MATERIALS AND METHODS: DNA vaccine was made by cloning the tyrosine hydroxylase minigene fused to the potato virus X coat protein gene into the expression vector. The A/J mice were vaccinated by three intramuscular injections. For immunogenicity study, immune response was estimated by target cells cytotoxicity assay, interferon-gamma production in enzyme-linked immunospot assay and antigen-specific antibodies in 14 days after the final vaccination. Antitumor effect was assessed by measurement of tumor volume and event-free survival rate in mice with engrafted NB41A3 murine neuroblastoma cells following three intramuscular injections of the vaccine: 7 days before, 5 and 10 days after tumor engraftment. The immune response was also assessed on the 30th day after tumor engraftment. RESULTS: The immunogenicity and antitumor effect of the vaccine in the form of aqueous solution of DNA and DNA-PEI conjugate were compared. Splenocytes cytotoxicity was the highest in the group of DNA-PEI vaccines (37.3 ± 6.9% lysis of target cells) compared with the unconjugated DNA vaccine (26.2 ± 4.0%) and placebo control (21.9 ± 3.7%). The production of interferon-gamma in the enzyme-linked immunospot assay was about ten times higher in the DNA-PEI group than in the other groups. The vaccine slowed or prevented the growth of the tumor. Mice vaccinated with the DNA-PEI vaccine had significantly better survival compared to control group (p < 0.0003). CONCLUSIONS: DNA vaccine against tyrosine hydroxylase, administered as a DNA-PEI 20 kDa conjugate, slows down the growth of neuroblastoma cells engrafted to mice.
Assuntos
Vacinas Anticâncer/farmacologia , Neuroblastoma/terapia , Vacinas Conjugadas/farmacologia , Vacinas de DNA/farmacologia , Animais , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , Feminino , Imunidade Celular/efeitos dos fármacos , Imunoterapia/métodos , Interferon gama/metabolismo , Masculino , Camundongos Endogâmicos , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/mortalidade , Neoplasias Experimentais/patologia , Neoplasias Experimentais/terapia , Neuroblastoma/imunologia , Neuroblastoma/mortalidade , Neuroblastoma/patologia , Polietilenoimina/química , Polietilenoimina/farmacologia , Vacinas Conjugadas/química , Vacinas Conjugadas/imunologia , Vacinas de DNA/química , Vacinas de DNA/imunologiaRESUMO
Biodegradable nanomaterials can protect antigens from degradation, promote cellular absorption, and enhance immune responses. We constructed a eukaryotic plasmid [pCAGGS-opti441-hemagglutinin (HA)] by inserting the optimized HA gene fragment of H9N2 AIV into the pCAGGS vector. The pCAGGS-opti441-HA/DGL was developed through packaging the pCAGGS-opti441-HA with dendrigraft poly-l-lysines (DGLs). DGL not only protected the pCAGGS-opti441-HA from degradation, but also exhibited high transfection efficiency. Strong cellular immune responses were induced in chickens immunized with the pCAGGS-opti441-HA/DGL. The levels of IFN-γ and IL-2, and lymphocyte transformation rate of the vaccinated chickens increased at the third week post the immunization. For the vaccinated chickens, T lymphocytes were activated and proliferated, the numbers of CD3+CD4+ and CD4+/CD8+ increased, and the chickens were protected completely against H9N2 AIV challenge. This study provides a method for the development of novel AIV vaccines, and a theoretical basis for the development of safe and efficient gene delivery carriers.
Assuntos
Anticorpos Antivirais/imunologia , Vacinas contra Influenza/farmacologia , Influenza Aviária/tratamento farmacológico , Vacinas de DNA/farmacologia , Animais , Anticorpos Antivirais/farmacologia , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Galinhas/imunologia , Galinhas/virologia , Vírus da Influenza A Subtipo H9N2/efeitos dos fármacos , Vírus da Influenza A Subtipo H9N2/imunologia , Vírus da Influenza A Subtipo H9N2/patogenicidade , Vacinas contra Influenza/química , Vacinas contra Influenza/genética , Vacinas contra Influenza/imunologia , Influenza Aviária/imunologia , Influenza Aviária/virologia , Polilisina/química , Polilisina/farmacologia , Vacinas de DNA/química , Vacinas de DNA/imunologiaRESUMO
DNA vaccinations are promising strategies for treating diseases that require cellular immunity (i.e., cancer and protozoan infection). Here, we report on the use of a liposomal nanocarrier (lipid nanoparticles (LNPs)) composed of an SS-cleavable and pH-activated lipidlike material (ssPalm) as an in vivo DNA vaccine. After subcutaneous administration, the LNPs containing an ssPalmE, an ssPalm with vitamin E scaffolds, elicited a higher gene expression activity in comparison with the other LNPs composed of the ssPalms with different hydrophobic scaffolds. Immunization with the ssPalmE-LNPs encapsulating plasmid DNA that encodes ovalbumin (OVA, a model tumor antigen) or profilin (TgPF, a potent antigen of Toxoplasma gondii) induced substantial antitumor or antiprotozoan effects, respectively. Flow cytometry analysis of the cells that had taken up the LNPs in draining lymph nodes (dLNs) showed that the ssPalmE-LNPs were largely taken up by macrophages and a small number of dendritic cells. We found that the transient deletion of CD169+ macrophages, a subpopulation of macrophages that play a key role in cancer immunity, unexpectedly enhanced the activity of the DNA vaccine. These data suggest that the ssPalmE-LNPs are effective DNA vaccine carriers, and a strategy for avoiding their being trapped by CD169+ macrophages will be a promising approach for developing next-generation DNA vaccines.
Assuntos
Lipídeos/química , Nanopartículas/química , Infecções por Protozoários/imunologia , Vacinas de DNA/química , Vacinas de DNA/imunologia , Vitamina E/imunologia , Animais , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , DNA/imunologia , Células Dendríticas/imunologia , Feminino , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Imunidade Celular/imunologia , Imunização/métodos , Lipossomos/química , Lipossomos/imunologia , Linfonodos/imunologia , Macrófagos/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Neoplasias/imunologia , Ovalbumina/imunologia , Plasmídeos/imunologia , Vitamina E/químicaRESUMO
The use of magnetic nanoparticles (MNPs), such as iron oxide nanoparticles (IONPs), in biomedicine is considered to be a valuable alternative to the more traditional materials due to their chemical stability, cost-effectiveness, surface functionalization, and the possibility to selectively attach and transport targeted species to the desired location under a magnetic field. One of the many main applications of MNPs is DNA separation, which enables genetic material manipulation; consequently, MNPs are used in numerous biotechnological methods, such as gene transfection and molecular recognition systems. In addition, the interaction between the surfaces of MNPs and DNA molecules and the magnetic nature of the resulting composite have facilitated the development of safe and effective gene delivery vectors to treat significant diseases, such as cancer and neurological disorders. Furthermore, the special recognition properties of nucleic acids based on the binding capacity of DNA and the magnetic behavior of the nanoparticles allowing magnetic separation and concentration of analytes have led to the development of biosensors and diagnostic assays; however, both of these applications face important challenges in terms of the improvement of selective nanocarriers and biosensing capacity. In this review, we discuss some aspects of the properties and surface functionalization of MNPs, the interactions between DNA and IONPs, the preparation of DNA nanoplatforms and their biotechnological applications, such as the magnetic separation of DNA, magnetofection, preparation of DNA vaccines, and molecular recognition tools.
Assuntos
DNA/química , Compostos Férricos/química , Nanopartículas de Magnetita/química , Nanomedicina , DNA/isolamento & purificação , Portadores de Fármacos/química , Humanos , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Vacinas de DNA/química , Vacinas de DNA/imunologiaRESUMO
Controlled release strategies of DNA vaccine hold promise for the design of in vivo vaccination platforms, yet the formulation and sustained delivery still pose a substantial challenge. In this study, we developed a novel hybrid dual-particulate delivery system, nanoparticle-in-microsphere (NIM), to integrate the advantages of nano-sized polymer/DNA polyplex with the sustained-release microsphere for DNA vaccine delivery. The nano-sized cores, consisting of polyethylene glycol-graft-polyethylenimine (PEG-g-PEI)/DNA polyplexes, were formulated into PLGA microspheres using a solid-in-oil-in-water (S/O/W) emulsion. The PEG block was used as stabilizing excipient to make DNA soluble and stable in organic solvent to prevent the inactivation of DNA at aqueous-organic interface during encapsulation. The fashion of DNA in dry solid state greatly increased the encapsulation efficiency of DNA in NIMs. This new formulation exhibited a burst release less than 15% and then sustain release close to zero-order kinetics in physiological environment. In addition, the microspheres showed pH-sensitivity and degraded faster in lysosomal compartments, which contributed to the accelerated intracellular release kinetics of DNA. Finally, intramuscular injection of NIMs encoding HIV proteins elicited distinct humoral and cellular immune response in mice at low dose. These results thus may aid NIM-based vaccination towards more extensive clinical evaluations.
Assuntos
Microesferas , Nanopartículas/química , Polietilenoglicóis/química , Polietilenoimina/análogos & derivados , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Vacinas de DNA/imunologia , Animais , Células COS , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Portadores de Fármacos/química , Portadores de Fármacos/toxicidade , HIV/genética , Concentração de Íons de Hidrogênio , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos/química , Plasmídeos/metabolismo , Polietilenoimina/química , Células RAW 264.7 , Transfecção , Vacinas de DNA/química , Vacinas de DNA/metabolismo , Proteínas Virais/genética , Proteínas Virais/imunologia , Proteínas Virais/metabolismoRESUMO
Vaccines and immunotherapies involve a variety of technologies and act through different mechanisms to achieve a common goal, which is to optimize the immune response against an antigen. The antigen could be a molecule expressed on a pathogen (e.g., a disease-causing bacterium, a virus or another microorganism), abnormal or damaged host cells (e.g., cancer cells), environmental agent (e.g., nicotine from a tobacco smoke), or an allergen (e.g., pollen or food protein). Immunogenic vaccines and therapies optimize the immune response to improve the eradication of the pathogen or damaged cells. In contrast, tolerogenic vaccines and therapies retrain or blunt the immune response to antigens, which are recognized by the immune system as harmful to the host. To optimize the immune response to either improve the immunogenicity or induce tolerance, researchers employ different routes of administration, antigen-delivery systems, and adjuvants. Nanocarriers and adjuvants are of particular interest to the fields of vaccines and immunotherapy as they allow for targeted delivery of the antigens and direct the immune response against these antigens in desirable direction (i.e., to either enhance immunogenicity or induce tolerance). Recently, nanoparticles gained particular attention as antigen carriers and adjuvants. This review focuses on a particular subclass of nanoparticles, which are made of nucleic acids, so-called nucleic acid nanoparticles or NANPs. Immunological properties of these novel materials and considerations for their clinical translation are discussed.
Assuntos
Imunoterapia , Nanopartículas , Ácidos Nucleicos , Nanomedicina Teranóstica , Vacinas de DNA , Adjuvantes Imunológicos , Animais , Sistemas de Liberação de Medicamentos , Humanos , Nanopartículas/química , Nanotecnologia , Ácidos Nucleicos/química , Pesquisa Translacional Biomédica , Vacinas de DNA/administração & dosagem , Vacinas de DNA/química , Vacinas de DNA/imunologiaRESUMO
Human papillomaviruses (HPV) are small, double-stranded DNA viruses that cause cervical cancer, the second most lethal cancer among women in the world. Currently, two vaccines are on the market for preventing HPV-caused cervical cancers and warts. Both are virus-like particle (VLP)-based vaccines. However, these vaccines have limitations; they are costly, have an invasive route of administration, require trained personnel to administer, need cold chain storage to preserve them, and most of all, they are preventive vaccines that do not have curative effects. Therefore, it is necessary to develop therapeutic HPV vaccines to facilitate the control of HPV-associated malignancies and to address all these issues. Recently there are DNA vaccines under investigation to prevent HPV. In general, DNA-based vaccines are better than or an excellent alternative to traditional vaccines since they can closely mimic live infections and can induce both antibody and cell-mediated immune responses. DNA vaccines involve the delivery of plasmid DNA (pDNA) which encodes the specific antigens. DNA vaccines have potential to be effective therapeutic tools against HPV infections. Combining the VLP-based and DNA-based vaccines can be highly effective as they can complement each other. VLP vaccines are more prone to mucosal immunity whereas DNA vaccines are more towards systemic immunity. In this article, we discuss an optimal formulation that will contain both type of vaccines, preventive and therapeutic. A film dosage form can be a good option which can be administered in buccal or sublingual routes for systemic action or in the vaginal area for local action to treat cervical cancer and to protect from future infection. Multiple vaccines in native form or in particulate form can be incorporated in film dosage forms. The film dosage form of vaccines can elicit both antibody-mediated (preventative) and cell-mediated (therapeutic) mechanisms. Film dosage forms are feasible to prepare for vaccine administration in the mouth cavity, GI tract, and vagina.
Assuntos
Sistemas de Liberação de Medicamentos , Papillomaviridae/efeitos dos fármacos , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/farmacologia , Neoplasias do Colo do Útero/prevenção & controle , Vacinas de DNA/farmacologia , Vacinas de Partículas Semelhantes a Vírus/química , Composição de Medicamentos , Feminino , Humanos , Vacinas contra Papillomavirus/administração & dosagem , Vacinas de DNA/químicaRESUMO
Spring viremia of carp virus (SVCV) has caused mass mortality in cyprinids, with case fatality rates of young fish up to 90%, resulting in enormous economic losses in the aquaculture industry. Immersion vaccination is considered as the most effective method for juvenile fish to combating disease, due to its convenience for mass vaccination and stress-free administration. However, immune responses following immersion vaccination are generally less robust and of shorter duration as those induced through intraperitoneal injection. Herein, to enhance the efficient of immersion vaccine, functionalized single-walled carbon nanotubes (SWCNTs) as carrier were used to manufacture immersion DNA vaccine system (SWCNTs-pEGFP-M) with chemical modification. Results showed that SWCNTs-pEGFP-M could enter into fish body via immersion administration and express antigen proteins in fish kidney and spleen. Moreover, stronger and longer duration immune responses (including serum antibody production and immune genes expression) can be induced in fish vaccinated with SWCNTs-pEGFP-M in comparison with those vaccinated with pEGFP-M alone. Notably, SWCNTs can increase the immune protective effect of naked DNA vaccine by ca. 23.8%. Altogether, this study demonstrates that SWCNTs as a promising DNA vaccine carrier might be used to vaccinate large-scale juvenile fish by bath administration approach, which can provide an outlook for future vaccination strategies against SVCV.
Assuntos
Doenças dos Peixes/prevenção & controle , Nanotubos de Carbono/química , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/imunologia , Vacinação/veterinária , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Aquicultura , Carpas , Portadores de Fármacos/química , Doenças dos Peixes/virologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Rim/metabolismo , Infecções por Rhabdoviridae/prevenção & controle , Infecções por Rhabdoviridae/virologia , Baço/metabolismo , Vacinação/métodos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/química , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Proteínas da Matriz Viral/genética , Proteínas da Matriz Viral/imunologia , Proteínas da Matriz Viral/metabolismo , Vacinas Virais/administração & dosagem , Vacinas Virais/química , Vacinas Virais/genéticaRESUMO
Castrate resistant prostate cancer (CRPC) remains a major challenge for healthcare professionals. Immunotherapeutic approaches, including DNA vaccination, hold the potential to harness the host's own immune system to mount a cell-mediated, anti-tumour response, capable of clearing disseminated tumour deposits. These anti-cancer vaccines represent a promising strategy for patients with advanced disease, however, to date DNA vaccines have demonstrated limited efficacy in clinical trials, owing to the lack of a suitable DNA delivery system. This study was designed to evaluate the efficacy of a two-tier delivery system incorporating cationic RALA/pDNA nanoparticles (NPs) into a dissolvable microneedle (MN) patch for the purposes of DNA vaccination against prostate cancer. Application of NP-loaded MN patches successfully resulted in endogenous production of the encoded Prostate Stem Cell Antigen (PSCA). Furthermore, immunisation with RALA/pPSCA loaded MNs elicited a tumour-specific immune response against TRAMP-C1 tumours ex vivo. Finally, vaccination with RALA/pPSCA loaded MNs demonstrated anti-tumour activity in both prophylactic and therapeutic prostate cancer models in vivo. This is further evidence that this two-tier MN delivery system is a robust platform for prostate cancer DNA vaccination. STATEMENT OF SIGNIFICANCE: This research describes the development and utilisation of our unique microneedle (MN) DNA delivery system, which enables penetration through the stratum corneum and deposition of the DNA within the highly immunogenic skin layers via a dissolvable MN matrix, and facilitates cellular uptake via complexation of pDNA cargo into nanoparticles (NPs) with the RALA delivery peptide. We report for the first time on using the NP-MN platform to immunise mice with encoded Prostate Stem Cell Antigen (mPSCA) for prostate cancer DNA vaccination. Application of the NP-MN system resulted in local mPSCA expression in vivo. Furthermore, immunisation with the NP-MN system induced a tumour-specific cellular immune response, and inhibited the growth of TRAMP-C1 prostate tumours in both prophylactic and therapeutic challenge models in vivo.
Assuntos
Antígenos de Neoplasias/imunologia , Vacinas Anticâncer , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Proteínas de Neoplasias/imunologia , Neoplasias de Próstata Resistentes à Castração , Vacinação , Vacinas de DNA , Animais , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/farmacologia , Linhagem Celular Tumoral , Proteínas Ligadas por GPI/imunologia , Células HEK293 , Humanos , Masculino , Camundongos , Agulhas , Neoplasias de Próstata Resistentes à Castração/imunologia , Neoplasias de Próstata Resistentes à Castração/patologia , Neoplasias de Próstata Resistentes à Castração/terapia , Vacinas de DNA/química , Vacinas de DNA/imunologia , Vacinas de DNA/farmacologiaRESUMO
Immunogenicity of DNA vaccines can be efficiently improved by adding adjuvants into their formulations. In this regard, the application of nano- and microparticles as vaccines adjuvants, or delivery systems, provides a powerful tool in designing modern vaccines. In the present study, we examined the role of "Supramolecular Hacky Sacks" (SHS) particles, made via the hierarchical self-assembly of a guanosine derivative, as a novel immunomodulator for DNA plasmid preparations. These plasmids code for the proteins HIV-1 Gag (pGag), the wild-type vaccinia virus Western Reserve A27 (pA27L), or a codon-optimized version of the latter (pOD1A27Lopt), which is also linked to the sequence of the outer domain-1 (OD1) from HIV-1 gp120 protein. We evaluated the enhancement of the immune responses generated by our DNA plasmid formulations in a murine model through ELISpot and ELISA assays. The SHS particles increased the frequencies of IFN-γ-producing cells in mice independently immunized with pGag and pA27L plasmids. Moreover, the addition of SHS to pGag and pA27L DNA plasmid formulations enhanced the production of IFN-γ (Th1-type) over IL-4 (Th2-type) cellular immune responses. Furthermore, pGag and pA27L plasmids formulated with SHS, triggered the production of antigen-specific IgG in mice, especially the IgG2a isotype. However, no improvement of either of those adaptive immune responses was observed in mice receiving pOD1A27Lopt+SHS. Here, we demonstrated that SHS particles have the ability to improve both arms of adaptive immunity of plasmid coding "wild-type" antigens without additional strategies to boost their immunogenicity. To the best of our knowledge, this is the first report of SHS guanosine-based particles as DNA plasmid adjuvants.
Assuntos
Guanosina , Nanopartículas , Plasmídeos , Vacinas de DNA/imunologia , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Animais , Especificidade de Anticorpos/imunologia , Citocinas/biossíntese , ELISPOT , Feminino , Guanosina/química , Anticorpos Anti-HIV/imunologia , HIV-1/imunologia , Humanos , Imunidade Humoral , Imunização , Imunogenicidade da Vacina , Imunoglobulina G/imunologia , Interferon gama/biossíntese , Camundongos , Nanopartículas/química , Plasmídeos/química , Plasmídeos/genética , Plasmídeos/imunologia , Vacinas de DNA/química , Vacinas de DNA/genéticaRESUMO
Lymphoid organs, which are populated by dendritic cells (DCs), are highly specialized tissues and provide an ideal microenvironment for T-cell priming. However, intramuscular or subcutaneous delivery of vaccine to DCs, a subset of antigen-presenting cells, has failed to stimulate optimal immune response for effective vaccination and need for adjuvants to induce immune response. To address this issue, we developed an in situ-forming injectable hybrid hydrogel that spontaneously assemble into microporous network upon subcutaneous administration, which provide a cellular niche to host immune cells, including DCs. In situ-forming injectable hybrid hydrogelators, composed of protein-polymer conjugates, formed a hydrogel depot at the close proximity to the dermis, resulting in a rapid migration of immune cells to the hydrogel boundary and infiltration to the microporous network. The biocompatibility of the watery microporous network allows recruitment of DCs without a DC enhancement factor, which was significantly higher than that of traditional hydrogel releasing chemoattractants, granulocyte-macrophage colony-stimulating factor. Owing to the sustained degradation of microporous hydrogel network, DNA vaccine release can be sustained, and the recruitment of DCs and their homing to lymph node can be modulated. Furthermore, immunization of a vaccine encoding amyloid-ß fusion proteinbearing microporous network induced a robust antigen-specific immune response in vivo and strong recall immune response was exhibited due to immunogenic memory. These hybrid hydrogels can be administered in a minimally invasive manner using hypodermic needle, bypassing the need for cytokine or DC enhancement factor and provide niche to host immune cells. These findings highlight the potential of hybrid hydrogels that may serve as a simple, yet multifunctional, platform for DNA vaccine delivery to modulate immune response.