Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine

BackgroundHIV remains a major public health issue in spite of antiretroviral therapy (ART). An innovative vaccine that can induce long-lasting and effective immunity is required to curb the persistently high numbers of new infections worldwide.MethodsA novel DNA vaccine was generated using a Simian-...

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Main Authors: Deepanwita Bose, Kenneth A. Rogers, Lisa M. Shirreff, Yahia Chebloune, Francois J. Villinger
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1481427/full
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author Deepanwita Bose
Kenneth A. Rogers
Lisa M. Shirreff
Yahia Chebloune
Francois J. Villinger
author_facet Deepanwita Bose
Kenneth A. Rogers
Lisa M. Shirreff
Yahia Chebloune
Francois J. Villinger
author_sort Deepanwita Bose
collection DOAJ
description BackgroundHIV remains a major public health issue in spite of antiretroviral therapy (ART). An innovative vaccine that can induce long-lasting and effective immunity is required to curb the persistently high numbers of new infections worldwide.MethodsA novel DNA vaccine was generated using a Simian-Human Immunodeficiency Virus (SHIV) backbone with a Zambian T/F clade C envelope and under the control of the caprine arthritis encephalitis virus long terminal repeats (LTRs) for constitutive expression. Due to the deleted integrase, this DNA vaccine “CSH-DIN-T/F Z331” performs only a single replication cycle. To increase immunogenicity, the co-expression of apoptotic genes (BAX, BAK, or caspase 8) incorporated at the end of Pol was tested to promote the release of apoptotic bodies taken up by dendritic cells leading to cross-presentation of antigen. The three vaccines (CSH-DIN-T/F Z331-BAX, CSH-DIN-T/F Z331-BAK, and CSH-DIN-T/F Z331-Cas8) were tested in vitro for expression and in vivo in BALB/cJ mice for immunogenicity.ResultsTransduced HEK293 cells co-cultured with CEMx174 confirmed the single replication cycle of the DNA vaccine and the induction of apoptosis by CSH-DIN-T/F Z331-Cas8 based on Annexin V expression. BALB/cJ mice were immunized with a combined intramuscular + intradermal/electroporation approach. Intracellular cytokine staining (ICS) from splenocytes collected 12 weeks post-prime/6 weeks post-boost demonstrated a clear superiority of caspase 8 expressing construct over the others, with higher proportions of IFN-γ-, IL-2-, and IL-21-producing CD8 T cells specific to Env, Gag, and Nef. The kinetics of immune response after various immunization schedules were also investigated.ConclusionThis novel single-cycle DNA vaccine with apoptotic genes demonstrated an enhanced immunogenicity primarily for antigen-specific CD8+ T-cell responses.
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spelling doaj-art-b282b2cfaaab4c9dbfdd00bd71abf4ff2025-08-20T03:08:43ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-04-011510.3389/fcimb.2025.14814271481427Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccineDeepanwita Bose0Kenneth A. Rogers1Lisa M. Shirreff2Yahia Chebloune3Francois J. Villinger4New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA, United StatesNew Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA, United StatesNew Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA, United StatesLaboratoire Pathogénèse et Vaccination Lentivirales (PAVAL) Lab., Institut National de Recherche d’Agriculture et Environnement, Université Grenoble Alpes, Saint Martin d’Hères, FranceNew Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA, United StatesBackgroundHIV remains a major public health issue in spite of antiretroviral therapy (ART). An innovative vaccine that can induce long-lasting and effective immunity is required to curb the persistently high numbers of new infections worldwide.MethodsA novel DNA vaccine was generated using a Simian-Human Immunodeficiency Virus (SHIV) backbone with a Zambian T/F clade C envelope and under the control of the caprine arthritis encephalitis virus long terminal repeats (LTRs) for constitutive expression. Due to the deleted integrase, this DNA vaccine “CSH-DIN-T/F Z331” performs only a single replication cycle. To increase immunogenicity, the co-expression of apoptotic genes (BAX, BAK, or caspase 8) incorporated at the end of Pol was tested to promote the release of apoptotic bodies taken up by dendritic cells leading to cross-presentation of antigen. The three vaccines (CSH-DIN-T/F Z331-BAX, CSH-DIN-T/F Z331-BAK, and CSH-DIN-T/F Z331-Cas8) were tested in vitro for expression and in vivo in BALB/cJ mice for immunogenicity.ResultsTransduced HEK293 cells co-cultured with CEMx174 confirmed the single replication cycle of the DNA vaccine and the induction of apoptosis by CSH-DIN-T/F Z331-Cas8 based on Annexin V expression. BALB/cJ mice were immunized with a combined intramuscular + intradermal/electroporation approach. Intracellular cytokine staining (ICS) from splenocytes collected 12 weeks post-prime/6 weeks post-boost demonstrated a clear superiority of caspase 8 expressing construct over the others, with higher proportions of IFN-γ-, IL-2-, and IL-21-producing CD8 T cells specific to Env, Gag, and Nef. The kinetics of immune response after various immunization schedules were also investigated.ConclusionThis novel single-cycle DNA vaccine with apoptotic genes demonstrated an enhanced immunogenicity primarily for antigen-specific CD8+ T-cell responses.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1481427/fullSHIVDNA vaccinelentivectorapoptotic geneimmune responsesmouse immunization
spellingShingle Deepanwita Bose
Kenneth A. Rogers
Lisa M. Shirreff
Yahia Chebloune
Francois J. Villinger
Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
Frontiers in Cellular and Infection Microbiology
SHIV
DNA vaccine
lentivector
apoptotic gene
immune responses
mouse immunization
title Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
title_full Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
title_fullStr Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
title_full_unstemmed Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
title_short Enhanced HIV immune responses elicited by an apoptotic single-cycle SHIV lentivector DNA vaccine
title_sort enhanced hiv immune responses elicited by an apoptotic single cycle shiv lentivector dna vaccine
topic SHIV
DNA vaccine
lentivector
apoptotic gene
immune responses
mouse immunization
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1481427/full
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