Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens

Alessandra Gallinaro, Martina Borghi, Roberta Bona, Felicia Grasso, Laura Calzoletti, Laura Palladino, Serena Cecchetti, Maria Fenicia Vescio, Daniele Macchia, Valeria Morante, Andrea Canitano, Nigel Temperton, Maria Rita Castrucci, Mirella Salvatore, Zuleika Michelini, Andrea Cara, Donatella Negri

Research output: Contribution to journalArticle

Abstract

Viral vectors represent an attractive technology for vaccine delivery. We exploited the integrase defective lentiviral vector (IDLV) as a platform for delivering relevant antigens within the context of the ADITEC collaborative research program. In particular, Influenza virus hemagglutinin (HA) and nucleoprotein (NP) were delivered by IDLVs while H1N1 A/California/7/2009 subunit vaccine (HAp) with or without adjuvant was used to compare the immune response in a murine model of immunization. In order to maximize the antibody response against HA, both IDLVs were also pseudotyped with HA (IDLV-HA/HA and IDLV-NP/HA, respectively). Groups of CB6F1 mice were immunized intramuscularly with a single dose of IDLV-NP/HA, IDLV-HA/HA, HAp alone, or with HAp together with the systemic adjuvant MF59. Six months after the vaccine prime all groups were boosted with HAp alone. Cellular and antibody responses to influenza antigens were measured at different time points after the immunizations. Mice immunized with HA-pseudotyped IDLVs showed similar levels of anti-H1N1 IgG over time, evaluated by ELISA, which were comparable to those induced by HAp + MF59 vaccination, but significantly higher than those induced by HAp alone. The boost with HAp alone induced an increase of antibodies in all groups, and the responses were maintained at higher levels up to 18 weeks post-boost. The antibody response was functional and persistent overtime, capable of neutralizing virus infectivity, as evaluated by hemagglutination inhibition and microneutralization assays. Moreover, since neuraminidase (NA)-expressing plasmid was included during IDLV preparation, immunization with IDLV-NP/HA and IDLV-HA/HA also induced functional anti-NA antibodies, evaluated by enzyme-linked lectin assay. IFNγ-ELISPOT showed evidence of HA-specific response in IDLV-HA/HA immunized animals and persistent NP-specific CD8+ T cell response in IDLV-NP/HA immunized mice. Taken together our results indicate that IDLV can be harnessed for producing a vaccine able to induce a comprehensive immune response, including functional antibodies directed toward HA and NA proteins present on the vector particles in addition to a functional T cell response directed to the protein transcribed from the vector.

Original languageEnglish
Pages (from-to)171
JournalFrontiers in Immunology
Volume9
DOIs
Publication statusPublished - 2018

Fingerprint

Integrases
Hemagglutinins
Human Influenza
Vaccines
Antigens
Nucleoproteins
Neuraminidase
Antibody Formation
Immunization
T-Lymphocytes
Enzyme-Linked Immunospot Assay
Subunit Vaccines

Keywords

  • Animals
  • Antibodies, Viral/blood
  • Disease Models, Animal
  • Drug Delivery Systems
  • Enzyme-Linked Immunospot Assay
  • Female
  • Genetic Vectors
  • Hemagglutinin Glycoproteins, Influenza Virus/administration & dosage
  • Immunoglobulin G/blood
  • Influenza Vaccines/administration & dosage
  • Integrases/genetics
  • Interferon-gamma
  • Lentivirus/genetics
  • Mice
  • Orthomyxoviridae Infections/immunology
  • Vaccination/methods
  • Viral Core Proteins/administration & dosage

Cite this

Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens. / Gallinaro, Alessandra; Borghi, Martina; Bona, Roberta; Grasso, Felicia; Calzoletti, Laura; Palladino, Laura; Cecchetti, Serena; Vescio, Maria Fenicia; Macchia, Daniele; Morante, Valeria; Canitano, Andrea; Temperton, Nigel; Castrucci, Maria Rita; Salvatore, Mirella; Michelini, Zuleika; Cara, Andrea; Negri, Donatella.

In: Frontiers in Immunology, Vol. 9, 2018, p. 171.

Research output: Contribution to journalArticle

Gallinaro, Alessandra ; Borghi, Martina ; Bona, Roberta ; Grasso, Felicia ; Calzoletti, Laura ; Palladino, Laura ; Cecchetti, Serena ; Vescio, Maria Fenicia ; Macchia, Daniele ; Morante, Valeria ; Canitano, Andrea ; Temperton, Nigel ; Castrucci, Maria Rita ; Salvatore, Mirella ; Michelini, Zuleika ; Cara, Andrea ; Negri, Donatella. / Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens. In: Frontiers in Immunology. 2018 ; Vol. 9. pp. 171.
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AU - Gallinaro, Alessandra

AU - Borghi, Martina

AU - Bona, Roberta

AU - Grasso, Felicia

AU - Calzoletti, Laura

AU - Palladino, Laura

AU - Cecchetti, Serena

AU - Vescio, Maria Fenicia

AU - Macchia, Daniele

AU - Morante, Valeria

AU - Canitano, Andrea

AU - Temperton, Nigel

AU - Castrucci, Maria Rita

AU - Salvatore, Mirella

AU - Michelini, Zuleika

AU - Cara, Andrea

AU - Negri, Donatella

PY - 2018

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N2 - Viral vectors represent an attractive technology for vaccine delivery. We exploited the integrase defective lentiviral vector (IDLV) as a platform for delivering relevant antigens within the context of the ADITEC collaborative research program. In particular, Influenza virus hemagglutinin (HA) and nucleoprotein (NP) were delivered by IDLVs while H1N1 A/California/7/2009 subunit vaccine (HAp) with or without adjuvant was used to compare the immune response in a murine model of immunization. In order to maximize the antibody response against HA, both IDLVs were also pseudotyped with HA (IDLV-HA/HA and IDLV-NP/HA, respectively). Groups of CB6F1 mice were immunized intramuscularly with a single dose of IDLV-NP/HA, IDLV-HA/HA, HAp alone, or with HAp together with the systemic adjuvant MF59. Six months after the vaccine prime all groups were boosted with HAp alone. Cellular and antibody responses to influenza antigens were measured at different time points after the immunizations. Mice immunized with HA-pseudotyped IDLVs showed similar levels of anti-H1N1 IgG over time, evaluated by ELISA, which were comparable to those induced by HAp + MF59 vaccination, but significantly higher than those induced by HAp alone. The boost with HAp alone induced an increase of antibodies in all groups, and the responses were maintained at higher levels up to 18 weeks post-boost. The antibody response was functional and persistent overtime, capable of neutralizing virus infectivity, as evaluated by hemagglutination inhibition and microneutralization assays. Moreover, since neuraminidase (NA)-expressing plasmid was included during IDLV preparation, immunization with IDLV-NP/HA and IDLV-HA/HA also induced functional anti-NA antibodies, evaluated by enzyme-linked lectin assay. IFNγ-ELISPOT showed evidence of HA-specific response in IDLV-HA/HA immunized animals and persistent NP-specific CD8+ T cell response in IDLV-NP/HA immunized mice. Taken together our results indicate that IDLV can be harnessed for producing a vaccine able to induce a comprehensive immune response, including functional antibodies directed toward HA and NA proteins present on the vector particles in addition to a functional T cell response directed to the protein transcribed from the vector.

AB - Viral vectors represent an attractive technology for vaccine delivery. We exploited the integrase defective lentiviral vector (IDLV) as a platform for delivering relevant antigens within the context of the ADITEC collaborative research program. In particular, Influenza virus hemagglutinin (HA) and nucleoprotein (NP) were delivered by IDLVs while H1N1 A/California/7/2009 subunit vaccine (HAp) with or without adjuvant was used to compare the immune response in a murine model of immunization. In order to maximize the antibody response against HA, both IDLVs were also pseudotyped with HA (IDLV-HA/HA and IDLV-NP/HA, respectively). Groups of CB6F1 mice were immunized intramuscularly with a single dose of IDLV-NP/HA, IDLV-HA/HA, HAp alone, or with HAp together with the systemic adjuvant MF59. Six months after the vaccine prime all groups were boosted with HAp alone. Cellular and antibody responses to influenza antigens were measured at different time points after the immunizations. Mice immunized with HA-pseudotyped IDLVs showed similar levels of anti-H1N1 IgG over time, evaluated by ELISA, which were comparable to those induced by HAp + MF59 vaccination, but significantly higher than those induced by HAp alone. The boost with HAp alone induced an increase of antibodies in all groups, and the responses were maintained at higher levels up to 18 weeks post-boost. The antibody response was functional and persistent overtime, capable of neutralizing virus infectivity, as evaluated by hemagglutination inhibition and microneutralization assays. Moreover, since neuraminidase (NA)-expressing plasmid was included during IDLV preparation, immunization with IDLV-NP/HA and IDLV-HA/HA also induced functional anti-NA antibodies, evaluated by enzyme-linked lectin assay. IFNγ-ELISPOT showed evidence of HA-specific response in IDLV-HA/HA immunized animals and persistent NP-specific CD8+ T cell response in IDLV-NP/HA immunized mice. Taken together our results indicate that IDLV can be harnessed for producing a vaccine able to induce a comprehensive immune response, including functional antibodies directed toward HA and NA proteins present on the vector particles in addition to a functional T cell response directed to the protein transcribed from the vector.

KW - Animals

KW - Antibodies, Viral/blood

KW - Disease Models, Animal

KW - Drug Delivery Systems

KW - Enzyme-Linked Immunospot Assay

KW - Female

KW - Genetic Vectors

KW - Hemagglutinin Glycoproteins, Influenza Virus/administration & dosage

KW - Immunoglobulin G/blood

KW - Influenza Vaccines/administration & dosage

KW - Integrases/genetics

KW - Interferon-gamma

KW - Lentivirus/genetics

KW - Mice

KW - Orthomyxoviridae Infections/immunology

KW - Vaccination/methods

KW - Viral Core Proteins/administration & dosage

U2 - 10.3389/fimmu.2018.00171

DO - 10.3389/fimmu.2018.00171

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VL - 9

SP - 171

JO - Frontiers in Immunology

JF - Frontiers in Immunology

SN - 1664-3224

ER -