This study was to prepare the chitosan-pDer p 2 nanoparticles and to investigate the effect of chitosan-DNA nanoparticles on immune response in mice by oral delivery of chitosan-DNA nanoparticles. The nanoparticles we...This study was to prepare the chitosan-pDer p 2 nanoparticles and to investigate the effect of chitosan-DNA nanoparticles on immune response in mice by oral delivery of chitosan-DNA nanoparticles. The nanoparticles were synthesized by complexing chitosan with plasmid DNA. The DNA was fully complexed into chitosan-DNA nanoparticles, suggesting a 100% encapsulation efficiency. Chitosan-DNA complex renders a significant protection of the plasmid. No effect on cell viability was observed in both cell types and average cell viability over 100% was obtained. Oral gene delivery with chitosan-DNA nanoparticles can generate a higher level expression of gene in vivo. Oral chitosan-pDer p 2 nanoparticles in BALB/c mice can induce IFN-γ in serum and prevent subsequent sensitization of Th2 cell-regulated specific IgE responses. The data indicate that the oral administration of chitosan-pDer p 2 nanoparticles results in the expression of Der p 2 in the epithelial cells of both stomach and small intestine and the induction of Th1-type immune response. Cellular ＆ Molecular Immunology.展开更多
Tumor vaccine is a promising strategy for cancer immunotherapy by introducing tumor antigens into the body to activate specific anti-tumor immune responses.Along with the technological breakthroughs in genetic enginee...Tumor vaccine is a promising strategy for cancer immunotherapy by introducing tumor antigens into the body to activate specific anti-tumor immune responses.Along with the technological breakthroughs in genetic engineering and delivery systems,messenger ribonucleic acid(mRNA)technology has achieved unprecedented development and application over the last few years,especially the emergency use authorizations of two mRNA vaccines during the COVID-19 pandemic,which has saved countless lives and makes the world witness the powerful efficacy of mRNA technology in vaccines.However,unlike infectious disease vaccines,which mainly induce humoral immunity,tumor vaccines also need to activate potent cellular immunity to control tumor growth,which creates a higher demand for mRNA delivery to the lymphatic organs and antigen-presenting cells(APCs).Here we review the existing bottlenecks of mRNA tumor vaccines and advanced nano-based strategies to overcome those challenges,as well as future considerations of mRNA tumor vaccines and their delivery systems.展开更多
Objective:To prepare and characterize polycaprolactone(PCL)nanoparticles loaded with sonicator fragmented(SLA)and freeze-thaw Leishmania antigens(FTLA)and to investigate the in vitro immunogenicity of antigen-encapsul...Objective:To prepare and characterize polycaprolactone(PCL)nanoparticles loaded with sonicator fragmented(SLA)and freeze-thaw Leishmania antigens(FTLA)and to investigate the in vitro immunogenicity of antigen-encapsulated nanoparticles with calcium phosphate adjuvant.Methods:The water/oil/water binary emulsion solvent evaporation method was used to synthesize antigen-loaded PCL nanoparticles.Particles were characterized by scanning electron microscopy and zeta potential measurements.Their cytotoxicity in J774 macrophages in vitro was determined by MTT analysis.In addition,the amount of nitric oxide and the level of cytokines produced by macrophages were determined by Griess reaction and ELISA method,respectively.The protective effect of the developed formulations was evaluated by determining the infection index percentage in macrophages infected with Leishmania infantum.Results:Compared to the control group,SLA PCL and FTLA PCL nanoparticles with calcium phosphate adjuvant induced a 6-and 7-fold increase in nitric oxide,respectively.Additionally,the vaccine formulations promoted the production of IFN-γand IL-12.SLA PCL and FTLA PCL nanoparticles combined with calcium phosphate adjuvant caused an approximately 13-and 11-fold reduction in infection index,respectively,compared to the control group.Conclusions:The encapsulation of antigens obtained by both sonication and freeze-thawing into PCL nanoparticles and the formulations with calcium phosphate adjuvant show strong in vitro immune stimulating properties.Therefore,PCL-based antigen delivery systems and calcium phosphate adjuvant are recommended as a potential vaccine candidate against leishmaniasis.展开更多
Cancer vaccines aimed at expanding the pool or increasing the activity of tumor-specific T cells against malignancies is an important immunotherapy modality that has been extensively pursued in the past decades. Howev...Cancer vaccines aimed at expanding the pool or increasing the activity of tumor-specific T cells against malignancies is an important immunotherapy modality that has been extensively pursued in the past decades. However, the clinical efficacy of cancer vaccines remains modest in comparison to other immunotherapies, such as checkpoint blockade and adoptive T cell therapy. This unsatisfactory performance is likely due to the suboptimal selection of tumor antigens for vaccine and inefficient delivery platform. Recently, vaccines designed to target cancer neoantigens have shown marked promise in both preclinical and early clinical studies. However, enormous challenges need to be overcome to develop a highly efficient and safe delivery strategy for targeting cancer vaccines to professional antigen-presenting cells and eliciting optimized immune response against cancers. To meet these challenges, biomaterials, particularly biomaterials that are designed to respond to certain environmental stimuli, termed as stimuli-responsive biomaterials, are being actively developed to precisely manipulate the trafficking and release of cancer vaccines in vivo for enhanced therapeutic efficacy and safety. In this mini review, we provide a brief overview of the recent advances in applying stimuli-responsive biomaterials in enhancing non-cellular cancer vaccines while focusing on the chemistry and material design with varied responsiveness. We also discuss the present challenges and opportunities in the field and provide a perspective for future directions.展开更多
Even with implementation of current influenza vaccines,influenza still claims up to 500,000 lives worldwide annually,indicating a need for a better vaccine strategy.We have developed a technology to generate unique S_...Even with implementation of current influenza vaccines,influenza still claims up to 500,000 lives worldwide annually,indicating a need for a better vaccine strategy.We have developed a technology to generate unique S_(60)-HA1 pseudovirus nanoparticles(PVNPs)that display the receptor-binding HA1 domains of influenza viruses.Each self-assembled S_(60)-HA1 PVNP consists of a T=1 icosahedral S_(60) nanoparticle that resembles the inner shell of norovirus capsid and 60 surface-displayed HA1 antigens that are excellent vaccine targets.Soluble S_(60)-HA1 PVNPs presenting HA1 antigens of H7N9 influenza virus subtypes have been produced efficiently in large amount.Their three-dimensional(3D)structures have been solved by cryogenic electron microscopy.The PVNP-displayed HA1 antigens react with HA-specific antibody,and retain authentic sialic acid binding specificity and hemagglutinate human erythrocytes.The PVNPs are highly immunogenic,eliciting high titers of HA1-specific antibodies in mice and the mouse sera strongly inhibited hemagglutinations of homologous and heterologous influenza virus HA proteins.Therefore,the S_(60)-HA1 PVNPs may provide useful reagents to study influenza viruses and offer a potential new vaccine tactic to fight the deadly influenza disease.展开更多
Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-s...Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-specific. The ability to tune the immune response is especially important in the immunosuppressive microenvironments of tumors. Because TLR9 is located intracellularly, CpG must be internalized by immune cells for functionality. Polyplexes can be self- assembled through electrostatics using DNA （anionic） condensed by a positively charged carrier. These structures improve cell delivery and have been widely explored for gene therapy. In contrast, here we use cationic poly （^-amino esters） （PBAEs） to assemble polyplexes from CpG as an adjuvant to target and improve immune stimulation in cells and mouse models. Polyplexes were formed over a range of PBAE：CpG ratios, resulting in a library of complexes with increasingly positive charge and stronger binding as PBAE：CpG ratio increased. Although higher PBAE：CpG ratios exhibited improved CpG uptake, lower ratios of PBAE：CpG--which condensed CpG more weakly, activated DCs and tumorspecific T cells more effectively. In a mouse melanoma model, polyplexes with lower binding affinities improved survival more effectively compared with higher binding affinities. These data demonstrate that altering the polyplex interaction strength impacts accessibility of CpG to TLRs in immune cells. Thus, physiochemical properties, particularly the interplay between charge, uptake, and affinity, play a key role in determining the nature and efficacy of the immune response generated. This insight identifies new design considerations that must be balanced for engineering effective immunotherapies and vaccines.展开更多
Lipid nanoparticle(LNP) is commonly used to deliver mRNA vaccines.Currently,LNP optimization primarily relies on screening ionizable lipids by traditional experiments which consumes intensive cost and time.Current stu...Lipid nanoparticle(LNP) is commonly used to deliver mRNA vaccines.Currently,LNP optimization primarily relies on screening ionizable lipids by traditional experiments which consumes intensive cost and time.Current study attempts to apply computational methods to accelerate the LNP development for mRNA vaccines.Firstly,325 data samples of mRNA vaccine LNP formulations with IgG titer were collected.The machine learning algorithm,lightGBM,was used to build a prediction model with good performance(R^(2)>0.87).More importantly,the critical substructures of ionizable lipids in LNPs were identified by the algorithm,which well agreed with published results.The animal experimental results showed that LNP using DLin-MC3-DMA(MC3) as ionizable lipid with an N/P ratio at 6:1 induced higher efficiency in mice than LNP with SM-102,which was consistent with the model prediction.Molecular dynamic modeling further investigated the molecular mechanism of LNPs used in the experiment.The result showed that the lipid molecules aggregated to form LNPs,and mRNA molecules twined around the LNPs.In summary,the machine learning predictive model for LNP-based mRNA vaccines was first developed,validated by experiments,and further integrated with molecular modeling.The prediction model can be used for virtual screening of LNP formulations in the future.展开更多
文摘This study was to prepare the chitosan-pDer p 2 nanoparticles and to investigate the effect of chitosan-DNA nanoparticles on immune response in mice by oral delivery of chitosan-DNA nanoparticles. The nanoparticles were synthesized by complexing chitosan with plasmid DNA. The DNA was fully complexed into chitosan-DNA nanoparticles, suggesting a 100% encapsulation efficiency. Chitosan-DNA complex renders a significant protection of the plasmid. No effect on cell viability was observed in both cell types and average cell viability over 100% was obtained. Oral gene delivery with chitosan-DNA nanoparticles can generate a higher level expression of gene in vivo. Oral chitosan-pDer p 2 nanoparticles in BALB/c mice can induce IFN-γ in serum and prevent subsequent sensitization of Th2 cell-regulated specific IgE responses. The data indicate that the oral administration of chitosan-pDer p 2 nanoparticles results in the expression of Der p 2 in the epithelial cells of both stomach and small intestine and the induction of Th1-type immune response. Cellular ＆ Molecular Immunology.
基金This work was supported by National Natural Science Funds of China(Nos.92059110 and 81872808)Development Fund for Shanghai Talents(No.2020090,China)+3 种基金Natural Science Foundation of Shanghai Science and Technology Innovation Plan(No.22ZR1414100,China)FDU 2025-Excellence Program Fund,Project Supported by Shanghai Municipal Science and Technology Major Project(No.2018SHZDZX01,China)ZJLab,Shanghai Post-doctoral Excellence Program(No.2033016,China)China Postdoctoral Science Foundation Funded Project(No.2023M730711).
文摘Tumor vaccine is a promising strategy for cancer immunotherapy by introducing tumor antigens into the body to activate specific anti-tumor immune responses.Along with the technological breakthroughs in genetic engineering and delivery systems,messenger ribonucleic acid(mRNA)technology has achieved unprecedented development and application over the last few years,especially the emergency use authorizations of two mRNA vaccines during the COVID-19 pandemic,which has saved countless lives and makes the world witness the powerful efficacy of mRNA technology in vaccines.However,unlike infectious disease vaccines,which mainly induce humoral immunity,tumor vaccines also need to activate potent cellular immunity to control tumor growth,which creates a higher demand for mRNA delivery to the lymphatic organs and antigen-presenting cells(APCs).Here we review the existing bottlenecks of mRNA tumor vaccines and advanced nano-based strategies to overcome those challenges,as well as future considerations of mRNA tumor vaccines and their delivery systems.
文摘Objective:To prepare and characterize polycaprolactone(PCL)nanoparticles loaded with sonicator fragmented(SLA)and freeze-thaw Leishmania antigens(FTLA)and to investigate the in vitro immunogenicity of antigen-encapsulated nanoparticles with calcium phosphate adjuvant.Methods:The water/oil/water binary emulsion solvent evaporation method was used to synthesize antigen-loaded PCL nanoparticles.Particles were characterized by scanning electron microscopy and zeta potential measurements.Their cytotoxicity in J774 macrophages in vitro was determined by MTT analysis.In addition,the amount of nitric oxide and the level of cytokines produced by macrophages were determined by Griess reaction and ELISA method,respectively.The protective effect of the developed formulations was evaluated by determining the infection index percentage in macrophages infected with Leishmania infantum.Results:Compared to the control group,SLA PCL and FTLA PCL nanoparticles with calcium phosphate adjuvant induced a 6-and 7-fold increase in nitric oxide,respectively.Additionally,the vaccine formulations promoted the production of IFN-γand IL-12.SLA PCL and FTLA PCL nanoparticles combined with calcium phosphate adjuvant caused an approximately 13-and 11-fold reduction in infection index,respectively,compared to the control group.Conclusions:The encapsulation of antigens obtained by both sonication and freeze-thawing into PCL nanoparticles and the formulations with calcium phosphate adjuvant show strong in vitro immune stimulating properties.Therefore,PCL-based antigen delivery systems and calcium phosphate adjuvant are recommended as a potential vaccine candidate against leishmaniasis.
基金This work was supported in part by the Foundation Pierre Mercier pour la science, ISREC Foundation with a donation from the Bateman Foundation, Swiss National Science Foundation （Project grant 315230 173243）, Novartis Foundation for medical-biological Research （17A058）, and the Ecole polytechnique federale de Lausanne （EPFL）.
文摘Cancer vaccines aimed at expanding the pool or increasing the activity of tumor-specific T cells against malignancies is an important immunotherapy modality that has been extensively pursued in the past decades. However, the clinical efficacy of cancer vaccines remains modest in comparison to other immunotherapies, such as checkpoint blockade and adoptive T cell therapy. This unsatisfactory performance is likely due to the suboptimal selection of tumor antigens for vaccine and inefficient delivery platform. Recently, vaccines designed to target cancer neoantigens have shown marked promise in both preclinical and early clinical studies. However, enormous challenges need to be overcome to develop a highly efficient and safe delivery strategy for targeting cancer vaccines to professional antigen-presenting cells and eliciting optimized immune response against cancers. To meet these challenges, biomaterials, particularly biomaterials that are designed to respond to certain environmental stimuli, termed as stimuli-responsive biomaterials, are being actively developed to precisely manipulate the trafficking and release of cancer vaccines in vivo for enhanced therapeutic efficacy and safety. In this mini review, we provide a brief overview of the recent advances in applying stimuli-responsive biomaterials in enhancing non-cellular cancer vaccines while focusing on the chemistry and material design with varied responsiveness. We also discuss the present challenges and opportunities in the field and provide a perspective for future directions.
基金The research described in this study was supported by the National Institute of Allergy and Infectious Diseases(NIAID,No.R56 AI148426-01A1 to M.T.)Cincinnati Children Hospital Medical Center(CCHMC,Innovation Funds 2018-2020,GAP Fund 2020-2021,and Research Innovation and Pilot Grant 2020-2021 to M.T.)+1 种基金the Center for Clinical and Translational Science and Training(CCTST)of the University of Cincinnati College of Medicine(Pilot Collaborative Studies Grant 2018-2019 to M.T.)that was supported by the National Center for Advancing Translational Sciences of the National Institutes of Health(No.UL1TR001425).
文摘Even with implementation of current influenza vaccines,influenza still claims up to 500,000 lives worldwide annually,indicating a need for a better vaccine strategy.We have developed a technology to generate unique S_(60)-HA1 pseudovirus nanoparticles(PVNPs)that display the receptor-binding HA1 domains of influenza viruses.Each self-assembled S_(60)-HA1 PVNP consists of a T=1 icosahedral S_(60) nanoparticle that resembles the inner shell of norovirus capsid and 60 surface-displayed HA1 antigens that are excellent vaccine targets.Soluble S_(60)-HA1 PVNPs presenting HA1 antigens of H7N9 influenza virus subtypes have been produced efficiently in large amount.Their three-dimensional(3D)structures have been solved by cryogenic electron microscopy.The PVNP-displayed HA1 antigens react with HA-specific antibody,and retain authentic sialic acid binding specificity and hemagglutinate human erythrocytes.The PVNPs are highly immunogenic,eliciting high titers of HA1-specific antibodies in mice and the mouse sera strongly inhibited hemagglutinations of homologous and heterologous influenza virus HA proteins.Therefore,the S_(60)-HA1 PVNPs may provide useful reagents to study influenza viruses and offer a potential new vaccine tactic to fight the deadly influenza disease.
文摘Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-specific. The ability to tune the immune response is especially important in the immunosuppressive microenvironments of tumors. Because TLR9 is located intracellularly, CpG must be internalized by immune cells for functionality. Polyplexes can be self- assembled through electrostatics using DNA （anionic） condensed by a positively charged carrier. These structures improve cell delivery and have been widely explored for gene therapy. In contrast, here we use cationic poly （^-amino esters） （PBAEs） to assemble polyplexes from CpG as an adjuvant to target and improve immune stimulation in cells and mouse models. Polyplexes were formed over a range of PBAE：CpG ratios, resulting in a library of complexes with increasingly positive charge and stronger binding as PBAE：CpG ratio increased. Although higher PBAE：CpG ratios exhibited improved CpG uptake, lower ratios of PBAE：CpG--which condensed CpG more weakly, activated DCs and tumorspecific T cells more effectively. In a mouse melanoma model, polyplexes with lower binding affinities improved survival more effectively compared with higher binding affinities. These data demonstrate that altering the polyplex interaction strength impacts accessibility of CpG to TLRs in immune cells. Thus, physiochemical properties, particularly the interplay between charge, uptake, and affinity, play a key role in determining the nature and efficacy of the immune response generated. This insight identifies new design considerations that must be balanced for engineering effective immunotherapies and vaccines.
基金financially supported by the University of Macao Research Grants (MYRG2020-00113-ICMS,China)。
文摘Lipid nanoparticle(LNP) is commonly used to deliver mRNA vaccines.Currently,LNP optimization primarily relies on screening ionizable lipids by traditional experiments which consumes intensive cost and time.Current study attempts to apply computational methods to accelerate the LNP development for mRNA vaccines.Firstly,325 data samples of mRNA vaccine LNP formulations with IgG titer were collected.The machine learning algorithm,lightGBM,was used to build a prediction model with good performance(R^(2)>0.87).More importantly,the critical substructures of ionizable lipids in LNPs were identified by the algorithm,which well agreed with published results.The animal experimental results showed that LNP using DLin-MC3-DMA(MC3) as ionizable lipid with an N/P ratio at 6:1 induced higher efficiency in mice than LNP with SM-102,which was consistent with the model prediction.Molecular dynamic modeling further investigated the molecular mechanism of LNPs used in the experiment.The result showed that the lipid molecules aggregated to form LNPs,and mRNA molecules twined around the LNPs.In summary,the machine learning predictive model for LNP-based mRNA vaccines was first developed,validated by experiments,and further integrated with molecular modeling.The prediction model can be used for virtual screening of LNP formulations in the future.
