Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice

Antonia Follenzi, Manuela Battaglia, Angelo Lombardo, Andrea Annoni, Maria Grazia Roncarolo, Luigi Naldini

Research output: Contribution to journalArticle

Abstract

Stable gene replacement by in vivo administration at lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immuno-competent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgena expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for tha gene therapy of hemophillas and other liver-based diseases.

Original languageEnglish
Pages (from-to)3700-3709
Number of pages10
JournalBlood
Volume103
Issue number10
DOIs
Publication statusPublished - May 15 2004

Fingerprint

Factor IX
Human engineering
Green Fluorescent Proteins
Transgenes
Hepatocytes
Liver
Gene transfer
Spleen
T-cells
Genes
T-Lymphocytes
Humoral Immunity
Cellular Immunity
Bone Marrow Cells
Genetic Therapy
Transgenic Mice
Antibody Formation
Liver Diseases
Animal Models
Gene therapy

ASJC Scopus subject areas

  • Hematology

Cite this

Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice. / Follenzi, Antonia; Battaglia, Manuela; Lombardo, Angelo; Annoni, Andrea; Roncarolo, Maria Grazia; Naldini, Luigi.

In: Blood, Vol. 103, No. 10, 15.05.2004, p. 3700-3709.

Research output: Contribution to journalArticle

@article{82f464235b2a4a74af07e2f2ba8d0b39,
title = "Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice",
abstract = "Stable gene replacement by in vivo administration at lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immuno-competent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgena expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for tha gene therapy of hemophillas and other liver-based diseases.",
author = "Antonia Follenzi and Manuela Battaglia and Angelo Lombardo and Andrea Annoni and Roncarolo, {Maria Grazia} and Luigi Naldini",
year = "2004",
month = "5",
day = "15",
doi = "10.1182/blood-2003-09-3217",
language = "English",
volume = "103",
pages = "3700--3709",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "10",

}

TY - JOUR

T1 - Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice

AU - Follenzi, Antonia

AU - Battaglia, Manuela

AU - Lombardo, Angelo

AU - Annoni, Andrea

AU - Roncarolo, Maria Grazia

AU - Naldini, Luigi

PY - 2004/5/15

Y1 - 2004/5/15

N2 - Stable gene replacement by in vivo administration at lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immuno-competent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgena expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for tha gene therapy of hemophillas and other liver-based diseases.

AB - Stable gene replacement by in vivo administration at lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immuno-competent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgena expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for tha gene therapy of hemophillas and other liver-based diseases.

UR - http://www.scopus.com/inward/record.url?scp=2342561729&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2342561729&partnerID=8YFLogxK

U2 - 10.1182/blood-2003-09-3217

DO - 10.1182/blood-2003-09-3217

M3 - Article

C2 - 14701690

AN - SCOPUS:2342561729

VL - 103

SP - 3700

EP - 3709

JO - Blood

JF - Blood

SN - 0006-4971

IS - 10

ER -