Animal Models of Systemic Lupus Erythematosus (SLE) and Ex Vivo Assay Design for Drug Discovery

Seavey, Matthew M.; Lu, Lily; Stump, Kristine

doi:10.1002/0471141755.ph0560s53

Cited by 18 publications

(22 citation statements)

References 61 publications

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“…Serum and urine were collected throughout the study. Assay design and other specifics concerning this model are described elsewhere (25). BWF1 study.…”

Section: Study Designsmentioning

confidence: 99%

“…All graphs show time as "day on study" with day on study starting at 212 d of age and ending at 310 d of age. Assay design and other specifics concerning this model are described elsewhere (25).…”

Section: Study Designsmentioning

confidence: 99%

“…The measurement of serum anti-dsDNA and anti-Smith Ab was performed by an in-house-developed custom ELISA assay described elsewhere (25). Chromatin-coated plates were purchased from Inova Diagnostics.…”

Section: Ana Elisa Assaysmentioning

confidence: 99%

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Depletion of Autoreactive Plasma Cells and Treatment of Lupus Nephritis in Mice Using CEP-33779, a Novel, Orally Active, Selective Inhibitor of JAK2

Lu¹,

Stump²,

Wallace³

et al. 2011

The Journal of Immunology

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Accumulating evidence suggests that autoreactive plasma cells play an important role in systemic lupus erythematosus (SLE). In addition, several proinflammatory cytokines promote autoreactive B cell maturation and autoantibody production. Hence, therapeutic targeting of such cytokine pathways using a selective JAK2 inhibitor, CEP-33779 (JAK2 enzyme IC50 = 1.3 nM; JAK3 enzyme IC50/JAK2 enzyme IC50 = 65-fold), was tested in two mouse models of SLE. Age-matched, MRL/lpr or BWF1 mice with established SLE or lupus nephritis, respectively, were treated orally with CEP-33779 at 30 mg/kg (MRL/lpr), 55 mg/kg or 100 mg/kg (MRL/lpr and BWF1). Studies included reference standard, dexamethasone (1.5 mg/kg; MRL/lpr), and cyclophosphamide (50 mg/kg; MRL/lpr and BWF1). Treatment with CEP-33779 extended survival and reduced splenomegaly/lymphomegaly. Several serum cytokines were significantly decreased upon treatment including IL-12, IL-17A, IFN-α, IL-1β, and TNF-α. Anti-nuclear Abs and frequencies of autoantigen-specific, Ab-secreting cells declined upon CEP-33779 treatment. Increased serum complement levels were associated with reduced renal JAK2 activity, histopathology, and spleen CD138+ plasma cells. The selective JAK2 inhibitor CEP-33779 was able to mitigate several immune parameters associated with SLE advancement, including the protection and treatment of mice with lupus nephritis. These data support the possibility of using potent, orally active, small-molecule inhibitors of JAK2 to treat the debilitative disease SLE.

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“…Serum and urine were collected throughout the study. Assay design and other specifics concerning this model are described elsewhere (25). BWF1 study.…”

Section: Study Designsmentioning

confidence: 99%

Section: Study Designsmentioning

confidence: 99%

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Depletion of Autoreactive Plasma Cells and Treatment of Lupus Nephritis in Mice Using CEP-33779, a Novel, Orally Active, Selective Inhibitor of JAK2

Lu¹,

Stump²,

Wallace³

et al. 2011

The Journal of Immunology

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“…Recently, researches demonstrated that abnormalities of B cells (such as B10 cells), T lymphocytes and macrophages have been considered as leading bases of SLE [4,5]. Current treatments for SLE relies on anti-malarials, steroids, cytotoxic drugs and immunosuppressive agents, which, in spite of their curative effects, lead to a good many deleterious side effects such as therapyresistant disease symptoms and infection [6]. The hallmark of SLE is characterized by the release of autoantibodies which react with self-nuclear and cytoplasmic antigens, resulting in immunologic attacks to body organs [7].…”

Section: Introductionmentioning

confidence: 99%

Roles of 1,25(OH)2D3 and Vitamin D Receptor in the Pathogenesis of Rheumatoid Arthritis and Systemic Lupus Erythematosus by Regulating the Activation of CD4+ T Cells and the PKCδ/ERK Signaling Pathway

Ding²,

Xiang³

et al. 2016

Cell Physiol Biochem

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Background/Aims: The study aims to elucidate the roles of 1,25(OH)₂D₃ and vitamin D receptor (VDR) in the pathogenesis of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) by regulating the activation of CD4⁺ T cells and the PKCδ/ERK signaling pathway. Methods: From January 2013 to December 2015, a total of 130 SLE patients, 137 RA patients and 130 healthy controls were selected in this study. Serum levels of 1,25(OH)₂D₃ and VDR mRNA expression were detected by ELISA and real-time fluorescence quantitative PCR (RT-qPCR). Density gradient centrifugation was performed to separate peripheral blood mononuclear cells (PBMCs). CD4⁺ T cells were separated using magnetic activated cell sorting (MACS). CD4⁺T cells in logarithmic growth phase were collected and assigned into 9 groups: the normal control group, the normal negative control (NC) group, the VDR siRNA group, the RA control group, the RA NC group, the VDR over-expressed RA group, the SLE control group, the SLE NC group, and the VDR over-expressed SLE group. The mRNA and protein expressions of VDR, PKCδ, ERK1/2, CD11a, CD70 and CD40L were detected by RT-qPCR and Western blotting. Bisulfite genomic sequencing was conducted to monitor the methylation status of CD11a, CD70 and CD40L. Results: Compared with healthy controls, serum 1,25(OH)₂D₃ level and VDR mRNA expression in peripheral blood were decreased in SLE patients and RA patients. With the increase of concentrations of 1,25(OH)₂D₃ treatment, the VDR mRNA expression and DNA methylation levels of CD11a, CD70 and CD40L were declined, while the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L were elevated in SLE, RA and normal CD4⁺T cells. Compared with the SLE contro, RA control, SLE NC and RA NC groups, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L decreased but DNA methylation levels of CD11a, CD70 and CD40L increased in the VDR over-expressed SLE group and VDR over-expressed RA group. However, compared with the normal control and normal NC groups, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L increased, but DNA methylation levels of CD11a, CD70 and CD40L decreased in the VDR siRNA group. Compared with the normal control group, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L increased, but DNA methylation levels of CD11a, CD70 and CD40L decreased in the SLE control and RA control groups. Conclusion: Our study provide evidence that 1,25(OH)₂D₃ and VDR could inhibit the activation of CD4⁺ T cells and suppress the immune response of SLE and RA through inhibiting PKCδ/ERK pathway and promoting DNA methylation of CD11a, CD70 and CD40L.

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“…Remove serum and store at -80 °C for later analysis of ANA by ELISA. Refer to previous reports for detailed ANA ELISA protocols 24,25 . Store serum in multiple 10-20 µl aliquots to minimize freeze/thaw cycles, and allow a greater number of future assays.…”

Section: Final Tissue Harvestmentioning

confidence: 99%

The bm12 Inducible Model of Systemic Lupus Erythematosus (SLE) in C57BL/6 Mice

Klarquist

Janssen

2015

JoVE

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Citation: Klarquist, J., Janssen, E.M. The bm12 Inducible Model of Systemic Lupus Erythematosus (SLE) in C57BL/6 Mice. J. Vis. Exp. (105), e53319, doi:10.3791/53319 (2015). AbstractSystemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical and immunological manifestations. Several spontaneous and inducible animal models mirror common components of human disease, including the bm12 transfer model. Upon transfer of bm12 splenocytes or purified CD4 T cells, C57BL/6 mice rapidly develop large frequencies of T follicular helper cells (Tfh), germinal center (GC) B cells, and plasma cells followed by high levels of circulating anti-nuclear antibodies. Since this model utilizes mice on a pure C57BL/6 background, researchers can quickly and easily study disease progression in transgenic or knockout mouse strains in a relatively short period of time. Here we describe protocols for the induction of the model and the quantitation Tfh, GC B cells, and plasma cells by multi-color flow cytometry. Importantly, these protocols can also be used to characterize disease in most mouse models of SLE and identify Tfh, GC B cells, and plasma cells in other disease models. Video LinkThe video component of this article can be found at

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Animal Models of Systemic Lupus Erythematosus (SLE) and Ex Vivo Assay Design for Drug Discovery

Cited by 18 publications

References 61 publications

Depletion of Autoreactive Plasma Cells and Treatment of Lupus Nephritis in Mice Using CEP-33779, a Novel, Orally Active, Selective Inhibitor of JAK2

Depletion of Autoreactive Plasma Cells and Treatment of Lupus Nephritis in Mice Using CEP-33779, a Novel, Orally Active, Selective Inhibitor of JAK2

Roles of 1,25(OH)2D3 and Vitamin D Receptor in the Pathogenesis of Rheumatoid Arthritis and Systemic Lupus Erythematosus by Regulating the Activation of CD4+ T Cells and the PKCδ/ERK Signaling Pathway

The bm12 Inducible Model of Systemic Lupus Erythematosus (SLE) in C57BL/6 Mice

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