Abstract
Lupus nephritis (LN) is an autoimmune disorder in which co-stimulatory signals have been involved. Here we tested a cholesterol-conjugated-anti-CD40-siRNA in dendritic cells (DC) in vitro and in a model of LPS to check its potency and tissue distribution. Then, we report the effects of Chol-siRNA in an experimental model of mice with established lupus nephritis. Our in vitro studies in DC show a 100% intracellular delivery of Chol-siRNA, with a significant reduction in CD40 after LPS stimuli. In vivo in ICR mice, the CD40-mRNA suppressive effects of our Chol-siRNA on renal tissue were remarkably sustained over a 5 days after a single preliminary dose of Chol-siRNA. The intra-peritoneal administration of Chol-siRNA to NZB/WF1 mice resulted in a reduction of anti-DNA antibody titers, and histopathological renal scores as compared to untreated animals. The higher dose of Chol-siRNA prevented the progression of proteinuria as effectively as cyclophosphamide, whereas the lower dose was as effective as CTLA4. Chol-siRNA markedly reduced insterstitial CD3+ and plasma cell infiltrates as well as glomerular deposits of IgG and C3. Circulating soluble CD40 and activated splenic lymphocyte subsets were also strikingly reduced by Chol-siRNA. Our data show the potency of our compound for the therapeutic use of anti-CD40-siRNA in human LN and other autoimmune disorders.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Albuminuria / immunology
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Albuminuria / metabolism
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Albuminuria / therapy
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Animals
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Antibodies, Antinuclear / blood
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CD40 Antigens / genetics*
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CD40 Antigens / metabolism
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Cell Survival
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Cells, Cultured
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Complement C3 / metabolism
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Cytokines / blood
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Dendritic Cells / metabolism
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Disease Progression
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Gene Expression
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Gene Knockdown Techniques
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Humans
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Immunoglobulin G / metabolism
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Kidney / immunology
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Kidney / metabolism
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Kidney / pathology
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Lipopolysaccharides / pharmacology
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Lupus Nephritis / immunology
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Lupus Nephritis / metabolism
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Lupus Nephritis / therapy*
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Male
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Mice
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Mice, Inbred ICR
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Plasma Cells / immunology
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RNA Interference*
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RNA, Small Interfering / genetics*
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Spleen / immunology
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Spleen / metabolism
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Transfection
Substances
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Antibodies, Antinuclear
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CD40 Antigens
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Complement C3
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Cytokines
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Immunoglobulin G
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Lipopolysaccharides
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RNA, Small Interfering
Grants and funding
This study was supported by grants from Instituto de Salud Carlos III/FIS (PS09/00897, PS09/00107, PI10/2991), European Union Grant SYSKID Framework Programme 7 (FP7). Nuria Bolaños is a technician from ‘Contratos de tecnico de apoyo en el sistema nacional de la salud’ financed by ISCIII. Elia Ripoll was a fellowship from IDIBELL and from Societat Catalana de Transplantament. Josep M. Aran and Immaculada Herrero are a researchers from ‘Programa Estabilización Investigadores’ financed by ISCIII and Dpt. Salut Generalitat Catalunya, and Ana Merino was supported by a contract from Insitituto de Salud Carlos III, programa Sara Borrell. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.