Intravenous Delivery of HIV-Based Lentiviral Vectors Preferentially Transduces F4/80+ and Ly-6C+ Cells in Spleen, Important Target Cells in Autoimmune Arthritis

Ben T Van Den Brand,Michael Kracht,Fons A J Van De Loo,Claire E J Waterborg,Eline A Vermeij,Miranda B Bennink,Wim B Van Den Berg,Onno J Arntz,Nathalie Labrecque

doi:10.1371/journal.pone.0055356

Ben T Van Den Brand, Michael Kracht + Show 7 more

Open Access

https://doi.org/10.1371/journal.pone.0055356

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Abstract

Antigen presenting cells (APCs) play an important role in arthritis and APC specific gene therapeutic targeting will enable intracellular modulation of cell activity. Viral mediated overexpression is a potent approach to achieve adequate transgene expression levels and lentivirus (LV) is useful for sustained expression in target cells. Therefore, we studied the feasibility of lentiviral mediated targeting of APCs in experimental arthritis. Third generation VSV-G pseudotyped self-inactivating (SIN)-LV were injected intravenously and spleen cells were analyzed with flow cytometry for green fluorescent protein (GFP) transgene expression and cell surface markers. Collagen-induced arthritis (CIA) was induced by immunization with bovine collagen type II in complete Freund's adjuvant. Effect on inflammation was monitored macroscopically and T-cell subsets in spleen were analyzed by flow cytometry. Synovium from arthritic knee joints were analyzed for proinflammatory cytokine expression. Lentiviruses injected via the tail vein preferentially infected the spleen and transduction peaks at day 10. A dose escalating study showed that 8% of all spleen cells were targeted and further analysis showed that predominantly Ly6C+ and F4/80+ cells in spleen were targeted by the LV. To study the feasibility of blocking TAK1-dependent pathways by this approach, a catalytically inactive mutant of TAK1 (TAK1-K63W) was overexpressed during CIA. LV-TAK1-K63W significantly reduced incidence and arthritis severity macroscopically. Further histological analysis showed a significant decrease in bone erosion in LV-TAK1-K63W treated animals. Moreover, systemic Th17 levels were decreased by LV-TAK1-K63W treatment in addition to diminished IL-6 and KC production in inflamed synovium. In conclusion, systemically delivered LV efficiently targets monocytes and macrophages in spleen that are involved in autoimmune arthritis. Moreover, this study confirms efficacy of TAK1 targeting in arthritis. This approach may provide a valuable tool in targeting splenic APCs, to unravel their role in autoimmune arthritis and to identify and validate APC specific therapeutic targets.

Highlights

Inflammatory diseases, such as rheumatoid arthritis, are characterized by infiltration of leukocytes into the inflamed tissue consisting of various immune cells from both the innate and adaptive immune system
Effective gene therapeutic treatment of collageninduced arthritis has been achieved by ectopic overexpression of suppressor of cytokine signaling 3 (SOCS3) in splenic antigen presenting cells (APCs) [5]
This study shows that intravenous delivery of lentiviral vectors enables evaluation of therapeutic targets in splenic monocyte/macrophage-like cells that are important cells in autoimmune diseases

Summary

Introduction

Inflammatory diseases, such as rheumatoid arthritis, are characterized by infiltration of leukocytes into the inflamed tissue consisting of various immune cells from both the innate and adaptive immune system. These studies shown that the spleen is one of the organs predominantly targeted by lentivirus injected intravenously [7]. We validated intravenous lentivirus injections into DBA1/J mice by determining expression kinetics, a dose escalating study, and analysis of targeted cells.

Results

Conclusion