Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients

<b>Background:</b> Hepatitis B virus (HBV) is a major cause of morbidity and mortality globally, and chronic infections are associated with cirrhosis and hepatocellular carcinoma. Issues with conventional treatments and vaccines mean there is a need for new therapeutic vaccines, which mu...

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Main Authors: Md Haroon Or Rashid, Fumihiko Yasui, Takahiro Sanada, Risa Kono, Tomoko Honda, Bouchra Kitab, Lipi Akter, Masashi Utsunomiya, Risa Sato, Osamu Yoshida, Yoichi Hiasa, Yasunori Oda, Yasumasa Goh, Takashi Miyazaki, Michinori Kohara, Kyoko Tsukiyama-Kohara
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Language:English
Published: MDPI AG 2025-04-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/5/464
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author Md Haroon Or Rashid
Fumihiko Yasui
Takahiro Sanada
Risa Kono
Tomoko Honda
Bouchra Kitab
Lipi Akter
Masashi Utsunomiya
Risa Sato
Osamu Yoshida
Yoichi Hiasa
Yasunori Oda
Yasumasa Goh
Takashi Miyazaki
Michinori Kohara
Kyoko Tsukiyama-Kohara
author_facet Md Haroon Or Rashid
Fumihiko Yasui
Takahiro Sanada
Risa Kono
Tomoko Honda
Bouchra Kitab
Lipi Akter
Masashi Utsunomiya
Risa Sato
Osamu Yoshida
Yoichi Hiasa
Yasunori Oda
Yasumasa Goh
Takashi Miyazaki
Michinori Kohara
Kyoko Tsukiyama-Kohara
author_sort Md Haroon Or Rashid
collection DOAJ
description <b>Background:</b> Hepatitis B virus (HBV) is a major cause of morbidity and mortality globally, and chronic infections are associated with cirrhosis and hepatocellular carcinoma. Issues with conventional treatments and vaccines mean there is a need for new therapeutic vaccines, which must elicit a strong and sustainable immune response. Here, we evaluated the immunogenicity of dual-antigen vaccines containing hybrid surface (hy-LHBs) and core (HBc) antigens, combined with a carboxyl-vinyl polymer (CVP) as a mucoadhesive excipient, following intranasal administration in mice. <b>Methods:</b> Mice were intranasally administered a mixed vaccine (10 µg of hy-LHBs and 2.5 or 10 µg of HBc) with or without a CVP excipient, and they were assessed for their immune response (levels of IgGs or IgA antibodies in an ELISA, IFN-γ level in splenocytes in an ELISpot assay, and cytokine/chemokine levels in a BioPlex assay). A protein stability assay was also conducted for vaccine formulations with and without excipients. <b>Results:</b> Significantly enhanced IgG production was noted targeting hy-LHBs and (less markedly) HBc at 10 µg/antigen, but only a non-significant elevation was noted with the vaccine containing 2.5 µg HBc. The BioPlex assay showed a significant increase in IL-2 (#00-07, 0B), IL-12(p40)(#00), eotaxin (#00), MIP1α (#00, #00-07, 0B), and MCP-1 (#00-07, 0B) in mice that received treatment compared to those of untreated mice. The endpoint titers of IgG1 and IgG2a were measured, which were higher with CVP excipients than without. From the IgG2a/IgG1 ratio, a higher IgG1 response was induced by CVPs to hy-LHBs and a higher IgG2a response was induced to HBc. Th2-dominant phenotype to hy-LHBs was induced with CVP#00 in an ELISpot assay. The highest anti-hy-LHBs antibody titer was noted with the conventional CVP#00 excipient. Consistent with these results, a higher amount of neutralizing antibodies of HBV was induced with CVP#00 treatment and followed by #00-03 and #14-00. <b>Conclusions:</b> We consider that the addition of CVP excipients to vaccine formulation enhances immunogenicity and HBV antigen stability for intranasal vaccines. This effect was seen for both humoral and cell-mediated immune responses, indicating the potential of CVPs as excipients in intranasal HBV vaccines.
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spelling doaj-art-bdf450317cd14286bf8550d06bb693852025-08-20T03:48:01ZengMDPI AGVaccines2076-393X2025-04-0113546410.3390/vaccines13050464Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer ExcipientsMd Haroon Or Rashid0Fumihiko Yasui1Takahiro Sanada2Risa Kono3Tomoko Honda4Bouchra Kitab5Lipi Akter6Masashi Utsunomiya7Risa Sato8Osamu Yoshida9Yoichi Hiasa10Yasunori Oda11Yasumasa Goh12Takashi Miyazaki13Michinori Kohara14Kyoko Tsukiyama-Kohara15Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, JapanDepartment of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, JapanDepartment of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, JapanDepartment of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, JapanDepartment of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, JapanJoint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, JapanJoint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, JapanJoint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, JapanJoint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, JapanDepartment of Gastroenterology and Metabology, Ehime University Graduate School of Medicine, Toon 791-0295, JapanDepartment of Gastroenterology and Metabology, Ehime University Graduate School of Medicine, Toon 791-0295, JapanBeacle Co., Ltd., Kyoto 606-8305, JapanBeacle Co., Ltd., Kyoto 606-8305, JapanToko Yakuhin Kogyo Co., Ltd., Toyama 930-0211, JapanDepartment of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, JapanJoint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, Japan<b>Background:</b> Hepatitis B virus (HBV) is a major cause of morbidity and mortality globally, and chronic infections are associated with cirrhosis and hepatocellular carcinoma. Issues with conventional treatments and vaccines mean there is a need for new therapeutic vaccines, which must elicit a strong and sustainable immune response. Here, we evaluated the immunogenicity of dual-antigen vaccines containing hybrid surface (hy-LHBs) and core (HBc) antigens, combined with a carboxyl-vinyl polymer (CVP) as a mucoadhesive excipient, following intranasal administration in mice. <b>Methods:</b> Mice were intranasally administered a mixed vaccine (10 µg of hy-LHBs and 2.5 or 10 µg of HBc) with or without a CVP excipient, and they were assessed for their immune response (levels of IgGs or IgA antibodies in an ELISA, IFN-γ level in splenocytes in an ELISpot assay, and cytokine/chemokine levels in a BioPlex assay). A protein stability assay was also conducted for vaccine formulations with and without excipients. <b>Results:</b> Significantly enhanced IgG production was noted targeting hy-LHBs and (less markedly) HBc at 10 µg/antigen, but only a non-significant elevation was noted with the vaccine containing 2.5 µg HBc. The BioPlex assay showed a significant increase in IL-2 (#00-07, 0B), IL-12(p40)(#00), eotaxin (#00), MIP1α (#00, #00-07, 0B), and MCP-1 (#00-07, 0B) in mice that received treatment compared to those of untreated mice. The endpoint titers of IgG1 and IgG2a were measured, which were higher with CVP excipients than without. From the IgG2a/IgG1 ratio, a higher IgG1 response was induced by CVPs to hy-LHBs and a higher IgG2a response was induced to HBc. Th2-dominant phenotype to hy-LHBs was induced with CVP#00 in an ELISpot assay. The highest anti-hy-LHBs antibody titer was noted with the conventional CVP#00 excipient. Consistent with these results, a higher amount of neutralizing antibodies of HBV was induced with CVP#00 treatment and followed by #00-03 and #14-00. <b>Conclusions:</b> We consider that the addition of CVP excipients to vaccine formulation enhances immunogenicity and HBV antigen stability for intranasal vaccines. This effect was seen for both humoral and cell-mediated immune responses, indicating the potential of CVPs as excipients in intranasal HBV vaccines.https://www.mdpi.com/2076-393X/13/5/464HBVCVPhy-LHBsHBc
spellingShingle Md Haroon Or Rashid
Fumihiko Yasui
Takahiro Sanada
Risa Kono
Tomoko Honda
Bouchra Kitab
Lipi Akter
Masashi Utsunomiya
Risa Sato
Osamu Yoshida
Yoichi Hiasa
Yasunori Oda
Yasumasa Goh
Takashi Miyazaki
Michinori Kohara
Kyoko Tsukiyama-Kohara
Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
Vaccines
HBV
CVP
hy-LHBs
HBc
title Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
title_full Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
title_fullStr Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
title_full_unstemmed Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
title_short Immunogenicity of an Intranasal Dual (Core and Surface)-Antigen Vaccine Against Hepatitis B Virus Enhanced by Carboxyl-Vinyl Polymer Excipients
title_sort immunogenicity of an intranasal dual core and surface antigen vaccine against hepatitis b virus enhanced by carboxyl vinyl polymer excipients
topic HBV
CVP
hy-LHBs
HBc
url https://www.mdpi.com/2076-393X/13/5/464
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