AAV Induced Lupus Nephritis In Mice
Systemic Lupus Erythematosus (SLE) is an autoimmune disease in which the immune system produces antibodies to cellular components of the body, leading to widespread inflammation and tissue damage. There are numerous murine models that have long been employed in an effort to understand the cellular and genetic requirements for SLE induction. The classic models of spontaneous lupus include the F1 hybrid between the New Zealand Black (NZB) and New Zealand White (NZW) mouse strains (NZB/W F1) and its derivatives, the MRL/lpr (fas signaling deficient), and BXSB/Yaa (with a duplicated gene called toll-like receptor 7 (TLR7) that is responsible for the autoimmunity phenotype) strains. All of these models portray their own iterations of lupus-like diseases with a subset of symptoms akin to those observed in human SLE, namely, autoantibody production (antibodies to double stranded DNA (dsDNA), lymphoid activation and hyperplasia, elevated proteinuria (excess protein in the urine which is a sign of kidney damage), and lupus nephritis. All these strains reliably develop spontaneous lupus nephritis. AAV induced model is a refinement of the spontaneous lupus NZB/W F1 model. Injecting NZB mice with mouse interferon alpha (mIFNa) adeno-associated virus (AAV) vectors synchronizes the spontaneous disease state associated with NZB/W mice resulting in an acceleration of the disease (from months to weeks) and a synchronization of disease state across animals (Moser, 2009; Kono, 2006; Theofilopoulos, 1985; Rudofsky, 1993; Morel, 1994; Liu, 2013)
NZB female mice arrive at 11 weeks of age. One week later, day 0, all mice are weighed, randomized by weight and injected with either LacZ-AAV of IFNa-AAV intravenously. The IFNa-AAV vehicle control group develops significant disease compared to LacZ-AAV vehicle group by histopathology. Cyclophosphamide and tofacitinib (100 mg/kg) treatments significantly suppress disease.
On day 0, all groups will be injected IV with one of the two AAVs. One group of non-disease control animals will be dosed with 100 uL of LacZ-AAV. The remaining groups of animals will be diseased, induced with mIFNa-AAV. These groups will be composed of at least one vehicle control group, one positive treatment control group (tofacitinib or cyclophosphamide) and a number of drug treatment groups. Treatment will start on day -1, day 0, or day +1, depending on whether prophylactic, developing or therapeutic dosing is desired.
- Developing (Prophylactic) – Begin dosing on study day -1 (or earlier) and continue until necropsy on day 432.
- Semi-Established (Prophylactic) – Begin dosing on study day 0 and continue until necropsy on day 43.
- Established (Therapeutic) – Begin dosing on study day +1 (or more) and continue until necropsy on day 43.
- Route of administration: SC, PO, IP, IV
- Body weight (baseline, d7, d14, d21, 28, d35, d42)
- Serum for dsDNA Titers (baseline, d14, 28, d42)
- Urine for ELISA (baseline, , d14, 28, d42)
Urine Protein Scoring Key:
0 = None.
1 = 1–30 mg/dL
2 = 31–99 mg/dL
3 = 100–299 mg/dL
4 = 300–1999 mg/dL
5 = ≥2000 mg/dL
Kidneys are evaluated using H&E stained slides for characteristic histopathologic changes of lupus-induced nephritis including glomerular diameter, crescent percentage, cortical tubular protein cast severity, interstitial inflammation, and vasculitis.
Sample Data (Click on Image to Enlarge):
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