Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice

Siva K. Gandhapudi; Hua Shi; Martin R. Ward; John Peyton Bush; Margarita Avdiushko; Karuna Sundarapandiyan; Lauren V. Wood; Mania Dorrani; Afsheen Fatima; Joe Dervan; Frank Bedu-Addo; Greg Conn; Ted M. Ross; Jerold G. Woodward

doi:10.3390/v15020432

Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice

Siva K. Gandhapudi, Hua Shi, Martin R. Ward, John Peyton Bush, Margarita Avdiushko, Karuna Sundarapandiyan, Lauren V. Wood, Mania Dorrani, Afsheen Fatima, Joe Dervan, Frank Bedu-Addo, Greg Conn, Ted M. Ross, Jerold G. Woodward

Research output: Contribution to journal › Article › peer-review

Abstract

Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) proteins, we demonstrated that R-DOTAP nanoparticles promoted strong cellular- and antibody-mediated immune responses in both monovalent and bivalent vaccines. R-DOTAP-based vaccines induced antigen-specific and polyfunctional CD8⁺ and CD4⁺ effector T cells and memory T cells, respectively. Antibody responses induced by R-DOTAP showed a balanced Th1/Th2 type immunity, neutralizing activity and protection of mice from challenge with live SARS-CoV2 or influenza viruses. R-DOTAP also facilitated significant dose sparing of the vaccine antigens. These studies demonstrate that R-DOTAP is an excellent immune stimulator for the production of next-generation subunit vaccines containing multiple recombinant proteins.

Original language	English
Article number	432
Journal	Viruses
Volume	15
Issue number	2
DOIs	https://doi.org/10.3390/v15020432
State	Published - Feb 2023

Bibliographical note

Funding Information:
This study was funded, in part, by the National Institute of Allergy and Infectious Diseases, a component of the NIH, Department of Health and Human Services, under contract 75N93019C00052 to T.M.R. and sub-contract to J.G.W., the National Center for Research Resources and the National Center for Advancing Translational Sciences, the National Institutes of Health, through Grant UL1TR001998 through Center for Clinical and Translational Science Pilot Funding to J.G.W. and S.K.G. and The PDS Biotechnology sponsored research grants to J.G.W. In addition, the flow cytometry work was partially funded by the University of Kentucky Flow Cytometry & Immune Monitoring core facility supported in part by the Office of the Vice President for Research, the Markey Cancer Center and an NCI Center Core Support Grant (P30 CA177558) to the University of Kentucky Markey Cancer Center. T.M.R. is also supported as an Eminent Scholar through the Georgia Research Alliance.

Funding Information:
J.G.W. has received research funding from PDS Biotechnology. J.G.W., S.K.G. and M.W. have received stock options from PDS Biotechnology. S.K.G., F.B.A., K.S., L.W., J.D. and G.C. are officers in PDS Biotechnology. The remaining authors declare no conflict of interest.

Publisher Copyright:
© 2023 by the authors.

Keywords

R-DOTAP
SARS-CoV2
immune stimulatory cationic lipids
influenza
vaccine adjuvants
vaccine induced T cell immunity

ASJC Scopus subject areas

Infectious Diseases
Virology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.3390/v15020432

Cite this

Gandhapudi, S. K., Shi, H., Ward, M. R., Bush, J. P., Avdiushko, M., Sundarapandiyan, K., Wood, L. V., Dorrani, M., Fatima, A., Dervan, J., Bedu-Addo, F., Conn, G., Ross, T. M., & Woodward, J. G. (2023). Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice. Viruses, 15(2), Article 432. https://doi.org/10.3390/v15020432

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title = "Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice",

abstract = "Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) proteins, we demonstrated that R-DOTAP nanoparticles promoted strong cellular- and antibody-mediated immune responses in both monovalent and bivalent vaccines. R-DOTAP-based vaccines induced antigen-specific and polyfunctional CD8+ and CD4+ effector T cells and memory T cells, respectively. Antibody responses induced by R-DOTAP showed a balanced Th1/Th2 type immunity, neutralizing activity and protection of mice from challenge with live SARS-CoV2 or influenza viruses. R-DOTAP also facilitated significant dose sparing of the vaccine antigens. These studies demonstrate that R-DOTAP is an excellent immune stimulator for the production of next-generation subunit vaccines containing multiple recombinant proteins.",

keywords = "R-DOTAP, SARS-CoV2, immune stimulatory cationic lipids, influenza, vaccine adjuvants, vaccine induced T cell immunity",

author = "Gandhapudi, {Siva K.} and Hua Shi and Ward, {Martin R.} and Bush, {John Peyton} and Margarita Avdiushko and Karuna Sundarapandiyan and Wood, {Lauren V.} and Mania Dorrani and Afsheen Fatima and Joe Dervan and Frank Bedu-Addo and Greg Conn and Ross, {Ted M.} and Woodward, {Jerold G.}",

note = "Funding Information: This study was funded, in part, by the National Institute of Allergy and Infectious Diseases, a component of the NIH, Department of Health and Human Services, under contract 75N93019C00052 to T.M.R. and sub-contract to J.G.W., the National Center for Research Resources and the National Center for Advancing Translational Sciences, the National Institutes of Health, through Grant UL1TR001998 through Center for Clinical and Translational Science Pilot Funding to J.G.W. and S.K.G. and The PDS Biotechnology sponsored research grants to J.G.W. In addition, the flow cytometry work was partially funded by the University of Kentucky Flow Cytometry & Immune Monitoring core facility supported in part by the Office of the Vice President for Research, the Markey Cancer Center and an NCI Center Core Support Grant (P30 CA177558) to the University of Kentucky Markey Cancer Center. T.M.R. is also supported as an Eminent Scholar through the Georgia Research Alliance. Funding Information: J.G.W. has received research funding from PDS Biotechnology. J.G.W., S.K.G. and M.W. have received stock options from PDS Biotechnology. S.K.G., F.B.A., K.S., L.W., J.D. and G.C. are officers in PDS Biotechnology. The remaining authors declare no conflict of interest. Publisher Copyright: {\textcopyright} 2023 by the authors.",

year = "2023",

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number = "2",

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Gandhapudi, SK, Shi, H, Ward, MR, Bush, JP, Avdiushko, M, Sundarapandiyan, K, Wood, LV, Dorrani, M, Fatima, A, Dervan, J, Bedu-Addo, F, Conn, G, Ross, TM & Woodward, JG 2023, 'Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice', Viruses, vol. 15, no. 2, 432. https://doi.org/10.3390/v15020432

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T1 - Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice

AU - Gandhapudi, Siva K.

AU - Shi, Hua

AU - Ward, Martin R.

AU - Bush, John Peyton

AU - Avdiushko, Margarita

AU - Sundarapandiyan, Karuna

AU - Wood, Lauren V.

AU - Dorrani, Mania

AU - Fatima, Afsheen

AU - Dervan, Joe

AU - Bedu-Addo, Frank

AU - Conn, Greg

AU - Ross, Ted M.

AU - Woodward, Jerold G.

N1 - Funding Information: This study was funded, in part, by the National Institute of Allergy and Infectious Diseases, a component of the NIH, Department of Health and Human Services, under contract 75N93019C00052 to T.M.R. and sub-contract to J.G.W., the National Center for Research Resources and the National Center for Advancing Translational Sciences, the National Institutes of Health, through Grant UL1TR001998 through Center for Clinical and Translational Science Pilot Funding to J.G.W. and S.K.G. and The PDS Biotechnology sponsored research grants to J.G.W. In addition, the flow cytometry work was partially funded by the University of Kentucky Flow Cytometry & Immune Monitoring core facility supported in part by the Office of the Vice President for Research, the Markey Cancer Center and an NCI Center Core Support Grant (P30 CA177558) to the University of Kentucky Markey Cancer Center. T.M.R. is also supported as an Eminent Scholar through the Georgia Research Alliance. Funding Information: J.G.W. has received research funding from PDS Biotechnology. J.G.W., S.K.G. and M.W. have received stock options from PDS Biotechnology. S.K.G., F.B.A., K.S., L.W., J.D. and G.C. are officers in PDS Biotechnology. The remaining authors declare no conflict of interest. Publisher Copyright: © 2023 by the authors.

PY - 2023/2

Y1 - 2023/2

N2 - Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) proteins, we demonstrated that R-DOTAP nanoparticles promoted strong cellular- and antibody-mediated immune responses in both monovalent and bivalent vaccines. R-DOTAP-based vaccines induced antigen-specific and polyfunctional CD8+ and CD4+ effector T cells and memory T cells, respectively. Antibody responses induced by R-DOTAP showed a balanced Th1/Th2 type immunity, neutralizing activity and protection of mice from challenge with live SARS-CoV2 or influenza viruses. R-DOTAP also facilitated significant dose sparing of the vaccine antigens. These studies demonstrate that R-DOTAP is an excellent immune stimulator for the production of next-generation subunit vaccines containing multiple recombinant proteins.

AB - Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) proteins, we demonstrated that R-DOTAP nanoparticles promoted strong cellular- and antibody-mediated immune responses in both monovalent and bivalent vaccines. R-DOTAP-based vaccines induced antigen-specific and polyfunctional CD8+ and CD4+ effector T cells and memory T cells, respectively. Antibody responses induced by R-DOTAP showed a balanced Th1/Th2 type immunity, neutralizing activity and protection of mice from challenge with live SARS-CoV2 or influenza viruses. R-DOTAP also facilitated significant dose sparing of the vaccine antigens. These studies demonstrate that R-DOTAP is an excellent immune stimulator for the production of next-generation subunit vaccines containing multiple recombinant proteins.

KW - R-DOTAP

KW - SARS-CoV2

KW - immune stimulatory cationic lipids

KW - influenza

KW - vaccine adjuvants

KW - vaccine induced T cell immunity

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SN - 1999-4915

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JO - Viruses

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Recombinant Protein Vaccines Formulated with Enantio-Specific Cationic Lipid R-DOTAP Induce Protective Cellular and Antibody-Mediated Immune Responses in Mice

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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