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. 2012 Oct 24;4(157):157ra141.
doi: 10.1126/scitranslmed.3004801.

NADPH oxidase inhibits the pathogenesis of systemic lupus erythematosus

Allison M Campbell  1 , Michael Kashgarian, Mark J Shlomchik
Affiliations

NADPH oxidase inhibits the pathogenesis of systemic lupus erythematosus

Allison M Campbell et al. Sci Transl Med. .

Abstract

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by loss of tolerance to self nucleic acids. The source of autoantigen that drives disease onset and progression is unclear. A candidate source of autoantigen is the neutrophil extracellular trap (NET), which releases nucleic acids into the extracellular environment, generating a structure composed of DNA coated with antimicrobial proteins. On the basis of in vitro and patient correlative studies, several groups have suggested that NETs may provide lupus autoantigens. The observation that NET release (NETosis) relies on activity of the phagocyte NADPH (reduced form of nicotinamide adenine dinucleotide phosphate) oxidase (Nox2) in neutrophils of both humans and mice provided a genetic strategy to test this hypothesis in vivo. Therefore, we crossed an X-linked nox2 null allele onto the lupus-prone MRL.Fas(lpr) genetic background and assessed immune activation, autoantibody generation, and SLE pathology. Counter to the prevailing hypothesis, Nox2-deficient lupus-prone mice had markedly exacerbated lupus, including increased spleen weight, increased renal disease, and elevated and altered autoantibody profiles. Moreover, heterozygous female mice, which have Nox2 deficiency in 50% of neutrophils, also had exacerbated lupus and altered autoantibody patterns, suggesting that failure to undergo normal Nox2-dependent cell death may result in release of immunogenic self-constituents that stimulate lupus. Our results indicate that NETosis does not contribute to SLE in vivo; instead, Nox2 acts to inhibit disease pathogenesis, making this enzyme an important target for further study and a candidate for therapeutic intervention.

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Figures

Fig. 1
Fig. 1
Increased spleen weight and expanded myeloid compartment in Nox2-deficient mice. (A) Spleen weight in grams as a function of nox2 genotype and sex (B–D) FACS analysis of percentage of CD11b+, F4/80+, Gr1int-lo macrophages (B), MHC II+, CD11c+ conventional dendritic cells (C), and Bst+, SiglecH+, pDCs (D) in the spleens of mice with the indicated genotype, reported as percentage of live. (E–F) FACS analysis of CD44+, CD62L−, activated CD4 T cells (E), and CD25+, FoxP3+ T cells (F) in the spleens of mice with the indicated genotype. Percentages are reported as a percentage of CD4+, TcRβ+ cells. Statistical analyses were performed with the one-tailed Mann Whitney test. Five or more mice were analyzed per genotype at 14 weeks of age.
Fig. 2
Fig. 2
Increased antibody forming cells in the absence of Nox2. (A) Gating strategy for AFCs in the spleen. Cells were initially gated on live, TcRβ− cells. CD44+ CD138+ cells (left) were then gated on intracellular kappa+ (right) to identify AFCs. (B–C) FACS quantification of TcRβ−, CD19low-int, CD44+, CD138+ intracellular kappa+ cells as a function of genotype for both percentage of live (B) and total numbers (C) in the spleen. (D–G) AFCs per spleen as determined by ELISPOT for IgK (D), IgG2a (E), IgG1 (F), and IgM (G). Statistical analyses were performed with the one-tailed Mann Whitney test. Five or more mice were analyzed per genotype. Mice were analyzed at 14 weeks of age.
Fig. 3
Fig. 3
Altered anti-self response in the absence of Nox2. (A) Representative ANA staining patterns from plasma of Nox2-sufficient (left) and Nox2-deficient animals (right). (B) Dominant ANA pattern quantitated for each genotype by dilution. (C) Intensity of cytoplasmic HEp-2 staining quantitated for each genotype by the dilution at which the dominant pattern disappeared. Statistical analysis was performed with the one-tailed Mann-Whitney test. (D–E) ELISA assessment of Anti-Sm antibodies (D) and anti-RNA antibodies (E) in the plasma of fully backcrossed mice of the indicated genotype. A one-tailed Chi-Squared test was used to determine statistical significance. Threshold positivity (indicated by dashed line) was set at 25,000 AU for Anti-Sm (D) or 8,000 AU for Anti-RNA (E). Samples were taken from 14-week old mice.
Fig. 4
Fig. 4
Increased renal disease in the absence of Nox2. (A) Proteinuria was assessed using Siemens Albustix(R). (B) Glomerulonephritis was scored blindly by M.K. on a scale of 1 to 6, and is represented as a function of Nox2 genotype in backcrossed mice. (C–F) Example of a Nox2-sufficient kidney (C and D) and deficient kidney (E and F) at 4X (left panels) and 10X magnification (right panels). Proteinuria assessment and histological analysis was performed on mice that were 14 weeks of age.
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References

    1. Christensen SR, Shupe J, Nickerson K, Kashgarian M, Flavell RA, Shlomchik MJ. Toll-like receptor 7 and TLR9 dictate autoantibody specificity and have opposing inflammatory and regulatory roles in a murine model of lupus. Immunity. 2006;25:417–428. - PubMed
    1. Nickerson KM, Christensen SR, Shupe J, Kashgarian M, Kim D, Elkon K, Shlomchik MJ. TLR9 regulates TLR7- and MyD88-dependent autoantibody production and disease in a murine model of lupus. J Immunol. 2010;184:1840–1848. - PMC - PubMed
    1. Nagata S, Hanayama R, Kawane K. Autoimmunity and the clearance of dead cells. Cell. 2010;140:619–630. - PubMed
    1. Botto M, Dell'Agnola C, Bygrave AE, Thompson EM, Cook HT, Petry F, Loos M, Pandolfi PP, Walport MJ. Homozygous C1q deficiency causes glomerulonephritis associated with multiple apoptotic bodies. Nat Genet. 1998;19:56–59. - PubMed
    1. Hanayama R, Tanaka M, Miyasaka K, Aozasa K, Koike M, Uchiyama Y, Nagata S. Autoimmune disease and impaired uptake of apoptotic cells in MFG-E8-deficient mice. Science. 2004;304:1147–1150. - PubMed

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