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Article: Enhanced Systemic Humoral Immune Response Induced in Mice by Generalized Modules for Membrane Antigens (GMMA) Is Associated with Affinity Maturation and Isotype Switching.

Piccioli, Diego / Buricchi, Francesca / Bacconi, Marta / Bechi, Nicoletta / Galli, Barbara / Ferlicca, Francesca / Luzzi, Enrico / Cartocci, Elena / Marchi, Sara / Romagnoli, Giacomo / Alfini, Renzo / Di Benedetto, Roberta / Gallorini, Simona / Savino, Silvana / Brunelli, Brunella / Bartolini, Erika / Micoli, Francesca

Vaccines

2023 Volume 11, Issue 7

Abstract: Generalized Modules for Membrane Antigens (GMMA) are outer membrane vesicles derived from Gram-negative bacteria that can be used to design affordable subunit vaccines. GMMA have been observed to induce a potent humoral immune response in preclinical and ...

Abstract	Generalized Modules for Membrane Antigens (GMMA) are outer membrane vesicles derived from Gram-negative bacteria that can be used to design affordable subunit vaccines. GMMA have been observed to induce a potent humoral immune response in preclinical and clinical studies. In addition, in preclinical studies, it has been found that GMMA can be exploited as optimal antigen carriers for both protein and saccharide antigens, as they are able to promote the enhancement of the antigen-specific humoral immune response when the antigen is overexpressed or chemically conjugated to GMMA. Here we investigated the mechanism of this GMMA carrier effect by immunizing mice and using factor H binding protein and GMMA of
Language	English
Publishing date	2023-07-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines11071219
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Article: Investigating the Impact of Delivery System Design on the Efficacy of Self-Amplifying RNA Vaccines.

Anderluzzi, Giulia / Lou, Gustavo / Gallorini, Simona / Brazzoli, Michela / Johnson, Russell / O'Hagan, Derek T / Baudner, Barbara C / Perrie, Yvonne

Vaccines

2020 Volume 8, Issue 2

Abstract: messenger RNA (mRNA)-based vaccines combine the positive attributes of both live-attenuated and subunit vaccines. In order for these to be applied for clinical use, they require to be formulated with delivery systems. However, there are limited in vivo ... ...

Abstract	messenger RNA (mRNA)-based vaccines combine the positive attributes of both live-attenuated and subunit vaccines. In order for these to be applied for clinical use, they require to be formulated with delivery systems. However, there are limited in vivo studies which compare different delivery platforms. Therefore, we have compared four different cationic platforms: (1) liposomes, (2) solid lipid nanoparticles (SLNs), (3) polymeric nanoparticles (NPs) and (4) emulsions, to deliver a self-amplifying mRNA (SAM) vaccine. All formulations contained either the non-ionizable cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or dimethyldioctadecylammonium bromide (DDA) and they were characterized in terms of physico-chemical attributes
Language	English
Publishing date	2020-05-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines8020212
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Article ; Online: The role of nanoparticle format and route of administration on self-amplifying mRNA vaccine potency.

Anderluzzi, Giulia / Lou, Gustavo / Woods, Stuart / Schmidt, Signe Tandrup / Gallorini, Simona / Brazzoli, Michela / Johnson, Russell / Roberts, Craig W / O'Hagan, Derek T / Baudner, Barbara C / Perrie, Yvonne

Journal of controlled release : official journal of the Controlled Release Society

2021 Volume 342, Page(s) 388–399

Abstract: The efficacy of RNA-based vaccines has been recently demonstrated, leading to the use of mRNA-based COVID-19 vaccines. The application of self-amplifying mRNA within these formulations may offer further enhancement to these vaccines, as self-amplifying ... ...

Abstract	The efficacy of RNA-based vaccines has been recently demonstrated, leading to the use of mRNA-based COVID-19 vaccines. The application of self-amplifying mRNA within these formulations may offer further enhancement to these vaccines, as self-amplifying mRNA replicons enable longer expression kinetics and more potent immune responses compared to non-amplifying mRNAs. To investigate the impact of administration route on RNA-vaccine potency, we investigated the immunogenicity of a self-amplifying mRNA encoding the rabies virus glycoprotein encapsulated in different nanoparticle platforms (solid lipid nanoparticles (SLNs), polymeric nanoparticles (PNPs) and lipid nanoparticles (LNPs)). These were administered via three different routes: intramuscular, intradermal and intranasal. Our studies in a mouse model show that the immunogenicity of our 4 different saRNA vaccine formulations after intramuscular or intradermal administration was initially comparable; however, ionizable LNPs gave higher long-term IgG responses. The clearance of all 4 of the nanoparticle formulations from the intramuscular or intradermal administration site was similar. In contrast, immune responses generated after intranasal was low and coupled with rapid clearance for the administration site, irrespective of the formulation. These results demonstrate that both the administration route and delivery system format dictate self-amplifying RNA vaccine efficacy.
MeSH term(s)	Animals ; COVID-19 ; COVID-19 Vaccines ; Humans ; Liposomes ; Mice ; Nanoparticles ; RNA, Messenger ; SARS-CoV-2 ; Vaccine Potency ; Vaccines, Synthetic ; mRNA Vaccines
Chemical Substances	COVID-19 Vaccines ; Lipid Nanoparticles ; Liposomes ; RNA, Messenger ; Vaccines, Synthetic ; mRNA Vaccines
Language	English
Publishing date	2021-12-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2021.12.008
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Article ; Online: Bacterial polysaccharides with zwitterionic charge motifs: Toll-like receptor 2 agonists, T cell antigens, or both?

Wack, Andreas / Gallorini, Simona

Immunopharmacology and immunotoxicology

2008 Volume 30, Issue 4, Page(s) 761–770

Abstract: Bacterial capsular polysaccharides (PS) which naturally contain zwitterionic charge motifs (ZPS) possess specific immunostimulatory activity, leading to direct activation of antigen-presenting cells (APCs) through Toll-like receptor 2 (TLR2) and of T ... ...

Abstract	Bacterial capsular polysaccharides (PS) which naturally contain zwitterionic charge motifs (ZPS) possess specific immunostimulatory activity, leading to direct activation of antigen-presenting cells (APCs) through Toll-like receptor 2 (TLR2) and of T cells in co-culture systems. When administered intraperitoneally, ZPS and bacteria expressing them are involved in the induction or regulation of T-cell dependent inflammatory processes such as intra-abdominal abscess formation. To generate vaccine candidates with antigen and adjuvant properties in one molecule we have chemically introduced zwitterionic motifs into naturally anionic PS and find that the resulting ZPS are TLR2 agonists, able to activate human and mouse APCs. Since T-regulatory cells and other T-cell subsets express TLR2, and TLR2 engagement modifies functionality and activation state of these cells, we speculate that most effects induced by natural and chemically derived ZPS may be explained by their TLR2 agonist properties, presumably through the combined action on TLR2-expressing APCs and T cells.
MeSH term(s)	Animals ; Antigens, Differentiation, T-Lymphocyte/physiology ; Antigens, T-Independent/physiology ; Cells, Cultured ; Coculture Techniques ; Epitopes, T-Lymphocyte/chemistry ; Epitopes, T-Lymphocyte/immunology ; Humans ; Polysaccharides, Bacterial/chemistry ; Polysaccharides, Bacterial/physiology ; Static Electricity ; Toll-Like Receptor 2/agonists ; Toll-Like Receptor 2/chemistry ; Toll-Like Receptor 2/physiology
Chemical Substances	Antigens, Differentiation, T-Lymphocyte ; Antigens, T-Independent ; Epitopes, T-Lymphocyte ; Polysaccharides, Bacterial ; Toll-Like Receptor 2
Language	English
Publishing date	2008-08-01
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	807033-7
ISSN	1532-2513 ; 0892-3973
ISSN (online)	1532-2513
ISSN	0892-3973
DOI	10.1080/08923970802279126
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Zs.A 1481: Show issues

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Article ; Online: Delivery of self-amplifying mRNA vaccines by cationic lipid nanoparticles: The impact of cationic lipid selection.

Lou, Gustavo / Anderluzzi, Giulia / Schmidt, Signe Tandrup / Woods, Stuart / Gallorini, Simona / Brazzoli, Michela / Giusti, Fabiola / Ferlenghi, Ilaria / Johnson, Russell N / Roberts, Craig W / O'Hagan, Derek T / Baudner, Barbara C / Perrie, Yvonne

Journal of controlled release : official journal of the Controlled Release Society

2020 Volume 325, Page(s) 370–379

Abstract: Self-amplifying RNA (SAM) represents a versatile tool that can be used to develop potent vaccines, potentially able to elicit strong antigen-specific humoral and cellular-mediated immune responses to virtually any infectious disease. To protect the SAM ... ...

Abstract	Self-amplifying RNA (SAM) represents a versatile tool that can be used to develop potent vaccines, potentially able to elicit strong antigen-specific humoral and cellular-mediated immune responses to virtually any infectious disease. To protect the SAM from degradation and achieve efficient delivery, lipid nanoparticles (LNPs), particularly those based on ionizable amino-lipids, are commonly adopted. Herein, we compared commonly available cationic lipids, which have been broadly used in clinical investigations, as an alternative to ionizable lipids. To this end, a SAM vaccine encoding the rabies virus glycoprotein (RVG) was used. The cationic lipids investigated included 3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol (DC-Chol), dimethyldioctadecylammonium (DDA), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) and N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-1-aminium (DOBAQ). Whilst all cationic LNP (cLNP) formulations promoted high association with cells in vitro, those formulations containing the fusogenic lipid 1,2-dioleoyl-sn-3-phosphoethanolamine (DOPE) in combination with DOTAP or DDA were the most efficient at inducing antigen expression. Therefore, DOTAP and DDA formulations were selected for further in vivo studies and were compared to benchmark ionizable LNPs (iLNPs). Biodistribution studies revealed that DDA-cLNPs remained longer at the injection site compared to DOTAP-cLNPs and iLNPs when administered intramuscularly in mice. Both the cLNP formulations and the iLNPs induced strong humoral and cellular-mediated immune responses in mice that were not significantly different at a 1.5 µg SAM dose. In summary, cLNPs based on DOTAP and DDA are an efficient alternative to iLNPs to deliver SAM vaccines.
MeSH term(s)	Animals ; Lipids ; Liposomes ; Mice ; Nanoparticles ; Quaternary Ammonium Compounds ; RNA, Messenger ; Tissue Distribution ; Vaccines
Chemical Substances	Lipids ; Liposomes ; Quaternary Ammonium Compounds ; RNA, Messenger ; Vaccines
Language	English
Publishing date	2020-07-01
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2020.06.027
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Zs.A 2055: Show issues

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Article ; Online: Rational Design of Adjuvant for Skin Delivery: Conjugation of Synthetic β-Glucan Dectin-1 Agonist to Protein Antigen.

Donadei, Agnese / Gallorini, Simona / Berti, Francesco / O'Hagan, Derek T / Adamo, Roberto / Baudner, Barbara C

Molecular pharmaceutics

2015 Volume 12, Issue 5, Page(s) 1662–1672

Abstract: The potential benefits of skin delivery of vaccines derive from the presence of a densely connected network of antigen presenting cells in the skin layer, most significantly represented by Langerhans cells and dermal dendritic cells. Targeting these ... ...

Abstract	The potential benefits of skin delivery of vaccines derive from the presence of a densely connected network of antigen presenting cells in the skin layer, most significantly represented by Langerhans cells and dermal dendritic cells. Targeting these cells by adjuvant conjugated to an antigen should result in enhanced immunogenicity of a vaccine. Since one of the most widely used adjuvants is an insoluble salt of aluminum (aluminum hydroxide) that cannot be used for skin delivery due to reactogenicity, we focused our attention on agonists of receptors present on skin dendritic cells, including the Dectin-1 receptor. β-(1-3)-glucans, which are the most abundant components of the fungal surface, are known to activate the innate immune response by interaction with the C-type lectin-like Dectin-1 receptor. In this work we identified by rational design a well-defined synthetic β-(1-3)-glucan hexasaccharide as a Dectin-1 agonist and chemically conjugated it to the genetically detoxified diphtheria toxin (CRM197) protein antigen, as a means to increase the binding to Dectin-1 receptor and to target to skin dendritic cells. We demonstrated that the in vitro activation of the receptor was significantly impacted by the presentation of the glucan on the protein carrier. In vivo results in mice showed that the conjugation of the synthetic β-(1-3)-glucan when delivered intradermally resulted in higher antibody titers in comparison to intramuscular (i.m.) immunization and was not different from subcutaneous (s.c.) delivery. These findings suggest that weak receptor binders can be turned into more potent agonists by the multivalent presentation of many ligands covalently conjugated to the protein core. Moreover, this approach is particularly valuable to increase the immunogenicity of antigens administered via skin delivery.
MeSH term(s)	Adjuvants, Immunologic/chemistry ; Animals ; Antigens/chemistry ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Lectins, C-Type/chemistry ; Mice ; Mice, Inbred BALB C ; beta-Glucans/agonists ; beta-Glucans/chemistry
Chemical Substances	Adjuvants, Immunologic ; Antigens ; Lectins, C-Type ; beta-Glucans ; dectin 1
Language	English
Publishing date	2015-05-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2138405-8
ISSN	1543-8392 ; 1543-8384
ISSN (online)	1543-8392
ISSN	1543-8384
DOI	10.1021/acs.molpharmaceut.5b00072
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Zs.A 6250: Show issues

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Article ; Online: Mannosylation of LNP Results in Improved Potency for Self-Amplifying RNA (SAM) Vaccines.

Goswami, Roshan / Chatzikleanthous, Despo / Lou, Gustavo / Giusti, Fabiola / Bonci, Alessandra / Taccone, Marianna / Brazzoli, Michela / Gallorini, Simona / Ferlenghi, Ilaria / Berti, Francesco / O'Hagan, Derek T / Pergola, Carlo / Baudner, Barbara C / Adamo, Roberto

ACS infectious diseases

2019 Volume 5, Issue 9, Page(s) 1546–1558

Abstract: Mannosylation of Lipid Nanoparticles (LNP) can potentially enhance uptake by Antigen Presenting Cells, which are highly abundant in dermal tissues, to improve the potency of Self Amplifying mRNA (SAM) vaccines in comparison to the established unmodified ... ...

Abstract	Mannosylation of Lipid Nanoparticles (LNP) can potentially enhance uptake by Antigen Presenting Cells, which are highly abundant in dermal tissues, to improve the potency of Self Amplifying mRNA (SAM) vaccines in comparison to the established unmodified LNP delivery system. In the current studies, we evaluated mannosylated LNP (MLNP), which were obtained by incorporation of a stable Mannose-cholesterol amine conjugate, for the delivery of an influenza (hemagglutinin) encoded SAM vaccine in mice, by both intramuscular and intradermal routes of administration. SAM MLNP exhibited
MeSH term(s)	Animals ; Bone Marrow Cells/cytology ; Bone Marrow Cells/virology ; Cells, Cultured ; Cholesterol/chemistry ; Dendritic Cells/cytology ; Dendritic Cells/virology ; Female ; Hemagglutinin Glycoproteins, Influenza Virus/administration & dosage ; Hemagglutinin Glycoproteins, Influenza Virus/genetics ; Hemagglutinin Glycoproteins, Influenza Virus/immunology ; Immunoglobulin G/metabolism ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/chemical synthesis ; Influenza Vaccines/genetics ; Influenza Vaccines/immunology ; Injections, Intradermal ; Mannose/chemistry ; Mice ; Nanoparticles ; Orthomyxoviridae Infections/immunology ; Orthomyxoviridae Infections/prevention & control ; Particle Size ; RNA, Messenger/administration & dosage ; RNA, Messenger/genetics ; RNA, Messenger/immunology
Chemical Substances	Hemagglutinin Glycoproteins, Influenza Virus ; Immunoglobulin G ; Influenza Vaccines ; RNA, Messenger ; Cholesterol (97C5T2UQ7J) ; Mannose (PHA4727WTP)
Language	English
Publishing date	2019-07-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2373-8227
ISSN (online)	2373-8227
DOI	10.1021/acsinfecdis.9b00084
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Article ; Online: Preparation of highly concentrated influenza vaccine for use in novel delivery approaches.

Kommareddy, Sushma / Bonificio, Amanda / Gallorini, Simona / Baudner, Barbara / Singh, Manmohan / O'Hagan, Derek

Journal of pharmaceutical sciences

2013 Volume 102, Issue 3, Page(s) 866–875

Abstract: Vaccine antigens are usually available only as dilute solutions, which are difficult to formulate into various novel delivery systems, which often require highly concentrated antigens. To address this problem, we have utilized tangential flow filtration ( ...

Abstract	Vaccine antigens are usually available only as dilute solutions, which are difficult to formulate into various novel delivery systems, which often require highly concentrated antigens. To address this problem, we have utilized tangential flow filtration (TFF), a simple and scalable process to prepare highly concentrated vaccine antigens. Here, we describe the optimization of TFF to concentrate hemagglutinin (HA) of egg-derived influenza antigens, from 2008 to 2009 seasonal vaccine, to concentrations up to 28 mg/mL. Concentrated antigen was evaluated by single radial immunodiffusion and reversed-phase high-performance liquid chromatographic analysis for the estimation of the HA content and a range of assays including size exclusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and bicinchoninic acid assay for protein characterization. In addition, the concentrated antigens retained their immunogenicity, confirmed by the induction of immune responses comparable to that of unprocessed antigen in a mouse model. The liquid concentrates were stable for up to 4 weeks, which could allow subsequent formulation into novel delivery technologies. Hence, we have used influenza HA to demonstrate that the fast, robust, and scalable approach of TFF can be used to concentrate antigens for use in novel delivery approaches. Moreover, the concentration process could be applicable for a variety of antigens and a wide range of novel vaccine delivery applications.
MeSH term(s)	Animals ; Antibody Formation ; Antigens, Viral/administration & dosage ; Antigens, Viral/immunology ; Antigens, Viral/isolation & purification ; Chromatography, Gel ; Drug Delivery Systems ; Electrophoresis, Polyacrylamide Gel ; Female ; Filtration/instrumentation ; Hemagglutinin Glycoproteins, Influenza Virus/administration & dosage ; Hemagglutinin Glycoproteins, Influenza Virus/immunology ; Hemagglutinin Glycoproteins, Influenza Virus/isolation & purification ; Humans ; Influenza A Virus, H1N1 Subtype/immunology ; Influenza A Virus, H3N2 Subtype/immunology ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Influenza Vaccines/isolation & purification ; Influenza, Human/immunology ; Influenza, Human/prevention & control ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections/immunology ; Orthomyxoviridae Infections/prevention & control
Chemical Substances	Antigens, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; Influenza Vaccines
Language	English
Publishing date	2013-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	3151-3
ISSN	1520-6017 ; 0022-3549
ISSN (online)	1520-6017
ISSN	0022-3549
DOI	10.1002/jps.23444
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Article ; Online: Fabrication of cell culture-derived influenza vaccine dissolvable microstructures and evaluation of immunogenicity in guinea pigs.

Bonificio, Amanda / Ghartey-Tagoe, Esi / Gallorini, Simona / Baudner, Barbara / Chen, Guohua / Singh, Parminder / O'Hagan, Derek T / Kommareddy, Sushma

Vaccine

2015 Volume 33, Issue 25, Page(s) 2930–2938

Abstract: Microstructure patches provide an opportunity for simple, effective, and safe vaccine administration, while achieving the desired immune response. We have evaluated the MicroCor transdermal system for cell culture-derived trivalent influenza vaccine ... ...

Abstract	Microstructure patches provide an opportunity for simple, effective, and safe vaccine administration, while achieving the desired immune response. We have evaluated the MicroCor transdermal system for cell culture-derived trivalent influenza vaccine administration. Influenza monovalent purified bulk vaccines (monobulks) (H1N1, H3N2, B) were concentrated by tangential flow filtration, lyophilized, and formulated with biocompatible excipients to form the microstructure array dissolvable tips. Standard single radial immunodiffusion (SRID) determined that the influenza antigens retained potency through the formulation and microstructure array fabrication processes. Array stability was evaluated for storage in both refrigerated and room temperature conditions. Microstructure mechanical strength was confirmed by application to excised pig skin, resulting in successful skin penetration and tip dissolution within 5 min of microstructure insertion. Guinea pigs immunized with influenza vaccine-loaded microstructures had hemagglutinin inhibition (HI) and IgG titers comparable to those obtained by intramuscular injection. After two immunizations, serum HI titers for all immunized groups were greater than 40 (>4-fold higher than the untreated group). These data demonstrate the feasibility for the development of skin delivery technologies that are compatible with cell culture-derived influenza vaccines.
MeSH term(s)	Administration, Cutaneous ; Animals ; Antibodies, Viral/blood ; Antigens, Viral/blood ; Antigens, Viral/immunology ; Cells, Cultured ; Female ; Guinea Pigs ; Hemagglutination Inhibition Tests ; Hemagglutinin Glycoproteins, Influenza Virus/immunology ; Humans ; Immunodiffusion ; Immunoglobulin G/blood ; Influenza A Virus, H1N1 Subtype/immunology ; Influenza A Virus, H3N2 Subtype/immunology ; Influenza B virus/immunology ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Influenza, Human/prevention & control ; Orthomyxoviridae Infections/prevention & control ; Vaccination
Chemical Substances	Antibodies, Viral ; Antigens, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; Immunoglobulin G ; Influenza Vaccines
Language	English
Publishing date	2015-06-09
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2015.04.059
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Zs.A 1930: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Influenza subunit vaccine coated microneedle patches elicit comparable immune responses to intramuscular injection in guinea pigs.

Kommareddy, S / Baudner, B C / Bonificio, A / Gallorini, S / Palladino, G / Determan, A S / Dohmeier, D M / Kroells, K D / Sternjohn, J R / Singh, M / Dormitzer, P R / Hansen, K J / O'Hagan, D T

Vaccine

2013 Volume 31, Issue 34, Page(s) 3435–3441

Abstract: Delivery of influenza vaccine using innovative approaches such as microneedles has been researched extensively in the past decade. In this study we present concentration followed by formulation and coating of monobulks from 2008/2009 seasonal vaccine on ... ...

Abstract	Delivery of influenza vaccine using innovative approaches such as microneedles has been researched extensively in the past decade. In this study we present concentration followed by formulation and coating of monobulks from 2008/2009 seasonal vaccine on to 3M's solid microstructured transdermal system (sMTS) by a GMP-scalable process. The hemagglutinin (HA) in monobulks was concentrated by tangential flow filtration (TFF) to achieve HA concentrations as high as 20mg/ml. The stability of the coated antigens was evaluated by the functional assay, single radial immunodiffusion (SRID). The data generated show stability of the coated antigen upon storage at 4°C and room temperature in the presence of desiccant for at least 8 weeks. Freeze-thaw stability data indicate the stability of the coated antigen in stressed conditions. The vaccine coated microstructures were evaluated in vivo in a guinea pig model, and resulted in immune titers comparable to the traditional trivalent vaccine administered intramuscularly. The data presented indicate the potential use of the technology in delivery of influenza vaccine. This paper also addresses the key issues of stability of coated antigen, reproducibility and scalability of the processes used in preparation of influenza vaccine coated microneedle patches that are important in developing a successful product.
MeSH term(s)	Administration, Cutaneous ; Animals ; Antibodies, Viral/blood ; Antigens, Viral/administration & dosage ; Antigens, Viral/immunology ; Drug Stability ; Female ; Guinea Pigs ; Hemagglutination Inhibition Tests ; Hemagglutinin Glycoproteins, Influenza Virus/administration & dosage ; Hemagglutinin Glycoproteins, Influenza Virus/immunology ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Injections, Intramuscular ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections/immunology ; Orthomyxoviridae Infections/prevention & control ; Reproducibility of Results ; Transdermal Patch ; Vaccination/instrumentation ; Vaccination/methods ; Vaccines, Subunit/administration & dosage ; Vaccines, Subunit/immunology
Chemical Substances	Antibodies, Viral ; Antigens, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; Influenza Vaccines ; Vaccines, Subunit
Language	English
Publishing date	2013-07-25
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2013.01.050
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