{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T05:04:37Z","timestamp":1784178277796,"version":"3.55.0"},"reference-count":68,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T00:00:00Z","timestamp":1783987200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"USDA","award":["AP23OA000000C007"],"award-info":[{"award-number":["AP23OA000000C007"]}]},{"name":"USDA","award":["2023-67015-39884"],"award-info":[{"award-number":["2023-67015-39884"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background: Nucleic acid vaccines adjuvanted with lipid nanoparticles (LNPs) are a simple and effective means of protection against infectious diseases through the delivery of antigen(s) that elicit(s) specific immunity. DNA vaccines are not commercially available for poultry mainly due to low protective efficacy and the associated production cost. Methods: We introduced here a novel nanoadjuvant system (namely, QTAP) comprising DOTAP and Quil-A for efficient delivery of plasmid DNA (pDNA) constructs into cells and boosting immune responses of chickens. Variable QTAP-pDNA-LNPs formulas were investigated for their physicochemical characteristics, cellular transfection, and in vivo efficacy in a vaccine\/challenge model using pDNA encoding both S and N proteins of avian Infectious Bronchitis Virus (IBV). Results: Under electron microscopy, all LNP formulas were spherical in shape and had an overall size range of 45.4\u2013376.3 nm. The formula with the smallest size had the lowest pDNA encapsulation and the highest polydispersity index (PDI). On the contrary, formulas with larger sizes and lower PDI values encapsulated a higher concentration of plasmid payload. Interestingly, the high-payload vaccine formula was stable at both 25 \u00b0C and 4 \u00b0C for up to 6 weeks of storage, but its gene expression dropped beyond this point at both temperatures. Following immunization of chickens, all groups were challenged with a virulent IBV. Surprisingly, both formulas were able to reduce the IBV viral shedding, indicating their effectiveness even when low concentrations of pDNA were incorporated in LNPs. However, pre-challenge sera from immunized chicks did not elicit IBV-specific antibodies, compared to birds immunized with a commercial live-attenuated IBV vaccine. Conclusions: This study suggests that the QTAP nanoadjuvant system is a safe and effective adjuvant for DNA immunization in poultry that could also be further developed for commercialization.<\/jats:p>","DOI":"10.3390\/vaccines14070613","type":"journal-article","created":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T08:04:31Z","timestamp":1784016271000},"page":"613","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Novel Effective Nanoadjuvant System for Poultry Vaccines"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3651-6039","authenticated-orcid":false,"given":"Fakry F.","family":"Mohamed","sequence":"first","affiliation":[{"name":"Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA"},{"name":"Department of Virology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44511, Sharkia, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5433-6939","authenticated-orcid":false,"given":"Hassanein H.","family":"Abozeid","sequence":"additional","affiliation":[{"name":"Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA"},{"name":"Department of Poultry Diseases, Faculty of Veterinary Medicine, Cairo University, Giza 12211, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matt","family":"Murray","sequence":"additional","affiliation":[{"name":"Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3305-7903","authenticated-orcid":false,"given":"Adel M.","family":"Talaat","sequence":"additional","affiliation":[{"name":"Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA"},{"name":"Vireo Vaccine International, Middleton 53562, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"424860","DOI":"10.1155\/2015\/424860","article-title":"Progress and Challenges toward the Development of Vaccines against Avian Infectious Bronchitis","volume":"2015","author":"Bande","year":"2015","journal-title":"J. Immunol. Res."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sanders, B., Koldijk, M., and Schuitemaker, H. (2014). Inactivated Viral Vaccines, in Vaccine Analysis: Strategies, Principles, and Control, Springer.","DOI":"10.1007\/978-3-662-45024-6_2"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1637\/aviandiseases-D-23-99998","article-title":"Pros and Cons on Use of Live Viral Vaccines in Commercial Chicken Flocks","volume":"67","author":"Ganapathy","year":"2023","journal-title":"Avian Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1144924","DOI":"10.1155\/2023\/1144924","article-title":"Global Emergence of Infectious Bronchitis Virus Variants: Evolution, Immunity, and Vaccination Challenges","volume":"2023","author":"Abozeid","year":"2023","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1038\/s41541-021-00292-w","article-title":"SARS-CoV-2 vaccines strategies: A comprehensive review of phase 3 candidates","volume":"6","author":"Kyriakidis","year":"2021","journal-title":"npj Vaccines"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.ymeth.2006.07.018","article-title":"Virus-like particles: Passport to immune recognition","volume":"40","author":"Grgacic","year":"2006","journal-title":"Methods"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1146\/annurev-immunol-032712-095910","article-title":"Pathways of Antigen Processing","volume":"31","author":"Blum","year":"2013","journal-title":"Annu. Rev. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"26","DOI":"10.11599\/germs.2013.1034","article-title":"DNA vaccines: Roles against diseases","volume":"3","author":"Khan","year":"2013","journal-title":"GERMS"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1306","DOI":"10.4161\/hv.28077","article-title":"Safety and efficacy of DNA vaccines: Plasmids vs. minicircles","volume":"10","author":"Stenler","year":"2014","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1186\/s13567-019-0698-z","article-title":"Recent advances in delivery of veterinary DNA vaccines against avian pathogens","volume":"50","author":"Jazayeri","year":"2019","journal-title":"Vet. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10-1128","DOI":"10.1128\/JVI.01016-20","article-title":"A Novel Mucosal Adjuvant System for Immunization against Avian Coronavirus Causing Infectious Bronchitis","volume":"94","author":"Chandrasekar","year":"2020","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.vaccine.2015.11.043","article-title":"DNA vaccination of poultry: The current status in 2015","volume":"34","author":"Meunier","year":"2016","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.4049\/jimmunol.2100909","article-title":"Development of a Nanoparticle Multiepitope DNA Vaccine against Virulent Infectious Bronchitis Virus Challenge","volume":"208","author":"Qin","year":"2022","journal-title":"J. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1586\/14760584.4.1.51","article-title":"DNA vaccines for poultry: The jump from theory to practice","volume":"4","author":"Haygreen","year":"2005","journal-title":"Expert Rev. Vaccines"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Kozak, M., and Hu, J. (2024). DNA Vaccines: Their Formulations, Engineering and Delivery. Vaccines, 12.","DOI":"10.3390\/vaccines12010071"},{"key":"ref_16","unstructured":"(2017). First DNA Vaccine Licensed for Chickens, Cision PR Newswire. Available online: https:\/\/www.prnewswire.com\/news-releases\/first-dna-vaccine-licensed-for-chickens-300554855.html."},{"key":"ref_17","unstructured":"Saif, Y.M. (2008). Infectious bronchitis. Diseases of Poultry, Wiley-Blackwell."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.meegid.2017.05.006","article-title":"Complete genome sequences of two avian infectious bronchitis viruses isolated in Egypt: Evidence for genetic drift and genetic recombination in the circulating viruses","volume":"53","author":"Abozeid","year":"2017","journal-title":"Infect. Genet. Evol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1080\/03079459708419194","article-title":"Relationship between sequence variation in the S1 spike","volume":"26","author":"Cavanagh","year":"1997","journal-title":"Avian Pathol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"363","DOI":"10.3390\/poultry2030027","article-title":"Infectious Bronchitis Virus (Gammacoronavirus) in poultry: Genomic architecture, post-translational modifications, and structural motifs","volume":"2","author":"Bhuiyan","year":"2023","journal-title":"Poultry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1585","DOI":"10.1292\/jvms.001585","article-title":"Multivalent DNA Vaccine Enhanced Protection Efficacy against Infectious Bronchitis Virus in Chickens","volume":"71","author":"Yang","year":"2009","journal-title":"J. Vet. Med. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0939-6411(02)00129-7","article-title":"The effect of co-administration of adjuvants with a nanoparticle-based genetic vaccine delivery system on the resulting immune responses","volume":"55","author":"Cui","year":"2003","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1615\/CritRevTherDrugCarrierSyst.2020033273","article-title":"Nanoparticles as Adjuvants in Vaccine Delivery","volume":"37","author":"Garg","year":"2020","journal-title":"Crit. Rev. Ther. Drug Carr. Syst."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Wu, Z., Yu, H., Wang, G., Liu, R., Ling, D., Ke, E., Liu, X., Xu, T., and Wang, S. (2026). Vaccination with Lipid Nanoparticle-Delivered VP2-DNA Elicits Immune Protection in Chickens Against Novel Variant Infectious Bursal Disease Virus (nVarIBDV). Vaccines, 14.","DOI":"10.3390\/vaccines14020113"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1186\/s12951-025-03668-5","article-title":"Lipid nanoparticles-mRNA based on the consensus sequences of avian coronavirus S1 and N genes protect animals against multiple viral infections","volume":"23","author":"Xiao","year":"2025","journal-title":"J. Nanobiotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.addr.2020.06.002","article-title":"Lipid nanoparticles for nucleic acid delivery: Current perspectives","volume":"154\u2013155","author":"Samaridou","year":"2020","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1021\/bc300381s","article-title":"The Headgroup Evolution of Cationic Lipids for Gene Delivery","volume":"24","author":"Zhi","year":"2013","journal-title":"Bioconjugate Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1002\/med.20090","article-title":"Cationic liposomes as non-viral carriers of gene medicines: Resolved issues, open questions, and future promises","volume":"27","author":"Karmali","year":"2006","journal-title":"Med. Res. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2307","DOI":"10.1039\/c1nr10166h","article-title":"The role of surface charge density in cationic liposome-promoted dendritic cell maturation and vaccine-induced immune responses","volume":"3","author":"Ma","year":"2011","journal-title":"Nanoscale"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Henson, T.R., Richards, K.A., Gandhapudi, S.K., Woodward, J.G., and Sant, A.J. (2023). R-DOTAP Cationic Lipid Nanoparticles Outperform Squalene-Based Adjuvant Systems in Elicitation of CD4 T Cells after Recombinant Influenza Hemagglutinin Vaccination. Viruses, 15.","DOI":"10.3390\/v15020538"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"102757","DOI":"10.1016\/j.cis.2022.102757","article-title":"Liposomes as biocompatible and smart delivery systems\u2014The current state","volume":"309","author":"Dymek","year":"2022","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.apsb.2023.10.012","article-title":"Lipid nanovehicles overcome barriers to systemic RNA delivery: Lipid components, fabrication methods, and rational design","volume":"14","author":"Yan","year":"2023","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1095","DOI":"10.1586\/erv.10.89","article-title":"Nano-microparticles as immune adjuvants: Correlating particle sizes and the resultant immune responses","volume":"9","author":"Oyewumi","year":"2010","journal-title":"Expert Rev. Vaccines"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1208\/s12249-022-02294-w","article-title":"Optimization of DOTAP\/chol Cationic Lipid Nanoparticles for mRNA, pDNA, and Oligonucleotide Delivery","volume":"23","author":"Sun","year":"2022","journal-title":"AAPS PharmSciTech"},{"key":"ref_35","first-page":"100197","article-title":"Effect of the amount of cationic lipid used to complex siRNA on the cytotoxicity and proinflammatory activity of siRNA-solid lipid nanoparticles","volume":"6","author":"Hanafy","year":"2023","journal-title":"Int. J. Pharm. X"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Touray, B.J., Hanafy, M., Phanse, Y., Hildebrand, R., and Talaat, A.M. (2023). Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1188754"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e202000786","DOI":"10.26508\/lsa.202000786","article-title":"TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells","volume":"3","author":"Bestle","year":"2020","journal-title":"Life Sci. Alliance"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 Years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.jviromet.2006.07.018","article-title":"Development and evaluation of a real-time Taqman RT-PCR assay for the detection of infectious bronchitis virus from infected chickens","volume":"138","author":"Callison","year":"2006","journal-title":"J. Virol. Methods"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"17617","DOI":"10.1038\/s41598-020-74091-z","article-title":"Codon optimization with deep learning to enhance protein expression","volume":"10","author":"Fu","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Qin, F., Xia, F., Chen, H., Cui, B., Feng, Y., Zhang, P., Chen, J., and Luo, M. (2021). A Guide to Nucleic Acid Vaccines in the Prevention and Treatment of Infectious Diseases and Cancers: From Basic Principles to Current Applications. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.633776"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.2460\/javma.1990.196.11.1851","article-title":"Risks associated with use of live vaccines","volume":"196","author":"Tizard","year":"1990","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Bodman-Harris, O., Rollier, C.S., and Iqbal, M. (2024). Approaches to Enhance the Potency of Vaccines in Chickens. Vaccines, 12.","DOI":"10.20944\/preprints202409.2319.v1"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"10-1128","DOI":"10.1128\/JVI.00667-21","article-title":"Identification of Novel T-Cell Epitopes on Infectious Bronchitis Virus N Protein and Development of a Multi-epitope Vaccine","volume":"95","author":"Qin","year":"2021","journal-title":"J. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7889","DOI":"10.1128\/jvi.71.10.7889-7894.1997","article-title":"The carboxyl-terminal 120-residue polypeptide of infectious bronchitis virus nucleocapsid induces cytotoxic T lymphocytes and protects chickens from acute infection","volume":"71","author":"Seo","year":"1997","journal-title":"J. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"11951","DOI":"10.1038\/s41598-018-30356-2","article-title":"A Recombinant Newcastle Disease Virus (NDV) Expressing S Protein of Infectious Bronchitis Virus (IBV) Protects Chickens against IBV and NDV","volume":"8","author":"Shirvani","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1637\/0005-2086(2003)047[0272:POCFIB]2.0.CO;2","article-title":"Protection of Chickens from Infectious Bronchitis by In Ovo and Intramuscular Vaccination with a DNA Vaccine Expressing the S1 Glycoprotein","volume":"47","author":"Kapczynski","year":"2003","journal-title":"Avian Dis."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Gupta, S.K., Dey, S., and Chellappa, M.M. (2016). DNA vaccination in chickens. Vaccine Design: Methods and Protocols, Volume 2: Vaccines for Veterinary Diseases, Springer.","DOI":"10.1007\/978-1-4939-3389-1_11"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"101495","DOI":"10.1016\/j.omtm.2025.101495","article-title":"Exploring the potential of saponins as adjuvants in lipid-nanoparticle-based mRNA vaccines","volume":"33","author":"Eygeris","year":"2025","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"107609","DOI":"10.1016\/j.ijid.2024.107609","article-title":"Harnessing Mosaic-Adjuvanted Nucleic Acid Nanovaccines for Enhanced Avian Influenza Immunity","volume":"152","author":"Touray","year":"2025","journal-title":"Int. J. Infect. Dis."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Catenacci, L., Rossi, R., Sechi, F., Buonocore, D., Sorrenti, M., Perteghella, S., Peviani, M., and Bonferoni, M.C. (2024). Effect of Lipid Nanoparticle Physico-Chemical Properties and Composition on Their Interaction with the Immune System. Pharmaceutics, 16.","DOI":"10.3390\/pharmaceutics16121521"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"113559","DOI":"10.1016\/j.jconrel.2025.02.055","article-title":"Investigating the stability of RNA-lipid nanoparticles in biological fluids: Unveiling its crucial role for understanding LNP performance","volume":"381","author":"Zhang","year":"2025","journal-title":"J. Control. Release"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1186\/s12951-025-03836-7","article-title":"Size control of lipid nanoparticles via simulation-based design of a microfluidic chip and its effect on mRNA delivery in vitro and in vivo","volume":"23","author":"Kim","year":"2025","journal-title":"J. Nanobiotechnol."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Tanaka, H., Miyama, R., Sakurai, Y., Tamagawa, S., Nakai, Y., Tange, K., Yoshioka, H., and Akita, H. (2021). Improvement of mRNA Delivery Efficiency to a T Cell Line by Modulating PEG-Lipid Content and Phospholipid Components of Lipid Nanoparticles. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13122097"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1021\/acs.accounts.1c00544","article-title":"Chemistry of Lipid Nanoparticles for RNA Delivery","volume":"55","author":"Eygeris","year":"2021","journal-title":"Acc. Chem. Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1186\/s12951-024-02972-w","article-title":"Navigating the intricate in-vivo journey of lipid nanoparticles tailored for the targeted delivery of RNA therapeutics: A quality-by-design approach","volume":"22","author":"Haghighi","year":"2024","journal-title":"J. Nanobiotechnol."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"P\u0142uciennik, K., Sici\u0144ska, P., Misztal, W., and Bukowska, B. (2024). Important Factors Affecting Induction of Cell Death, Oxidative Stress and DNA Damage by Nano- and Microplastic Particles In Vitro. Cells, 13.","DOI":"10.3390\/cells13090768"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1038\/s41541-026-01476-y","article-title":"Superior protection against tuberculosis using heterologous mRNA nanoadjuvant vaccines","volume":"11","author":"Touray","year":"2026","journal-title":"npj Vaccines"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/s13567-019-0631-5","article-title":"Development of a recombinant Newcastle disease virus-vectored vaccine for infectious bronchitis virus variant strains circulating in Egypt","volume":"50","author":"Abozeid","year":"2019","journal-title":"Vet. Res."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Sander, V.A., Corigliano, M.G., and Clemente, M. (2019). Promising Plant-Derived Adjuvants in the Development of Coccidial Vaccines. Front. Vet. Sci., 6.","DOI":"10.3389\/fvets.2019.00020"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1787","DOI":"10.1016\/j.vaccine.2009.01.091","article-title":"Advances in saponin-based adjuvants","volume":"27","author":"Sun","year":"2009","journal-title":"Vaccine"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"3752","DOI":"10.1016\/j.vaccine.2007.01.111","article-title":"Vaccine adjuvants revisited","volume":"25","author":"Aguilar","year":"2007","journal-title":"Vaccine"},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Morais, V., Suarez, N., Cibulski, S., and Silveira, F. (2025). Leaf Saponins of Quillaja brasiliensis as Powerful Vaccine Adjuvants. Pharmaceutics, 17.","DOI":"10.3390\/pharmaceutics17080966"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1007\/s00705-019-04273-2","article-title":"Adjuvant activity of multimolecular complexes based on Glycyrrhiza glabra saponins, lipids, and influenza virus glycoproteins","volume":"164","author":"Alexyuk","year":"2019","journal-title":"Arch. Virol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.addr.2016.01.022","article-title":"The role of helper lipids in lipid nanoparticles (LNPs) designed for oligonucleotide delivery","volume":"99","author":"Cheng","year":"2016","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Marques, R., Lacerda, R., and Rom\u00e3o, L. (2022). Internal Ribosome Entry Site (IRES)-Mediated Translation and Its Potential for Novel mRNA-Based Therapy Development. Biomedicines, 10.","DOI":"10.3390\/biomedicines10081865"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1586\/14760584.1.1.85","article-title":"Technological advances to increase immunogenicity of DNA vaccines","volume":"1","author":"Lemieux","year":"2002","journal-title":"Expert Rev. Vaccines"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"65","DOI":"10.3844\/ajisp.2009.65.83","article-title":"Cytokines as Adjuvants for Vaccine and Cellular Therapies for Cancer","volume":"5","author":"Capitini","year":"2009","journal-title":"Am. J. Immunol."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/7\/613\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T04:31:35Z","timestamp":1784176295000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/7\/613"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,14]]},"references-count":68,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["vaccines14070613"],"URL":"https:\/\/doi.org\/10.3390\/vaccines14070613","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,14]]}}}