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Departments of
*
Medicine and
Pathology, University of Southern California School of Medicine, Los Angeles, CA 90089; and
Department of Medicine, University of Florida, Gainesville, FL 32610
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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. In contrast, Stat6-deficient NZM mice display a significant reduction in incidence of kidney disease, with a dramatic increase in survival, despite the presence of high levels of anti-dsDNA Abs. The lack of correlation between levels of these autoantibodies and kidney disease raises the question of the direct cause-effect relationships between the presence of autoantibodies and kidney disease. Furthermore, these results also question the apparent equation of the effect of Stat deficiency with loss of secretion or response to particular cytokines. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Glomerulonephritis (GN; including glomerular hypercellularity, glomerular immune complex deposition, and proteinuria) is one of the major clinical hallmarks of SLE. In addition, there are characteristic serological abnormalities, some highly specific for the diagnosis of SLE (e.g., anti-dsDNA Abs, anti-Sm Abs), and other lupus-associated but not pathognomonic, of SLE (e.g., antinuclear Abs, anti-nRNP, anti-chromatin). The deposition of pathogenic autoantibodies in target organs such as kidneys implies the participation of autoantibody producing B lymphocytes (1). In addition, various cytokines, including IFN- and IL-4, have been suggested to be involved in the pathogenesis of the autoimmune disease in human and mouse models of SLE (7, 8, 9, 10, 11).
Stat proteins are transcription factors that provide a direct link between the cytokine receptors and cytokine-induced gene transcription (12). Stat4 and Stat6 are members of this family that are essential in mediating responses to IL-12 and IL-4, respectively (12, 13).
In the present study, we have produced NZM 2328 deficient in Stat4 or Stat6 and have used these new genetic models to address questions related to mechanisms important in the induction and perpetuation of autoimmune disease in SLE.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The original Stat4 and Stat6 null mice on a 129, C57BL/6 mixed background were kindly provided to us by Dr. M. Grusby (Harvard University School of Public Health, Boston, MA). The NZM2328 are maintained at the University of Southern California Laboratory Animal Resource Center (Los Angeles, CA). The Stat4- and Stat6-deficient mice were crossed onto the NZM2328 background, using a marker-assisted selection strategy. Each backcross generation was screened with polymorphic microsatellite markers at a density of 
25 cM. In each generation 25–30 males were screened, and the progeny carrying the target gene in combination with the lowest content of donor genes (129/C57BL/6) was chosen for breeding with NZM 2328 females to obtain the next backcross generation. For identification of Stat4 and Stat6 null mutations, PCR-based assays were used. One assay amplified the linked neomycin resistance (neo) gene using the primers 5'-GGT GAG ATG ACA GGA GAT C-3' and 5'-GAC TGG GCA CAA CAG ACA ATC-3'. The other assay amplified a segment of the respective Stat gene spanning the null-generating mutation using the following primers: 5'-GTA GAT CAG GAA GGT AGC TC-3' and 5'-CTG CCA GAG AGT CTT TTC C-3' (for Stat4) and 5'-CTC AGA GTC GCT AAA GCG G-3' and 5'-GGC AGT GGT TTG ATG GTG-3' (for Stat6).
In vitro culture
Pooled spleen cells from control and Stat4- or Stat6-deficient mice were activated with 1 µg/ml plate-bound anti-CD3 (145-2C11). After 1 wk in culture, cells were washed and restimulated with anti-CD3 for 24 h. Where indicated, IL-12 was added at 200 U/ml. Supernatants were harvested, and IFN- and IL-4 production were quantified using commercial ELISA kits per the manufacturer’s instructions.
For measuring NK cell cytolytic activity, splenocytes were enriched for NK cells (stained with biotin-anti-CD3 and rat anti mouse Ig and depleted of T and B lymphocytes using magnetic beads), and incubated with rIL-12 (200 U/ml) overnight. In control experiments, NK-enriched splenocytes were incubated with 250 µg/ml poly I:C. Cells were collected, washed, and plated with 51Cr-labeled YAC-1 target cells at various E:T ratios in a 4-h chromatin release assay. YAC-1 cells were labeled with 150 µCi sodium [51Cr] chromate (Amersham, Arlington Heights, IL) for 2 h at 37°C. Cells were washed three times and plated at 10-4 cells per well containing appropriately diluted effector cells. After 4 h, plates were spun down and supernatants were counted in a scintillation counter.
Flow cytometry
Splenocytes were obtained from control and Stat4- and Stat6-deficient mice and enriched for small resting B cells by treatment of splenocytes with anti-Thy 1.2 and complement followed by Percoll gradient centrifugation. Resting B cells were cultured with IL-4 (30 U/ml). After 48 h, cells were stained with FITC-anti-CD23 and biotinylated 10.2.16 anti-I-Az Ab (kindly provided by Dr. B. Kotzin, University of Colorado, Denver, CO). Flow cytometry was performed on a FACScan (BD Biosciences, San Jose, CA).
Serologic analysis
Serum Ig profile. Total IgG and IgG subclasses (IgG1, 2a, 2b, 3) were determined by ELISA as described (14). Microtiter plate wells were coated with Fc-specific F(ab')2 goat anti-mouse IgG (Southern Biotechnology Associates, Birmingham, AL). Bound IgG subclasses were measured using alkaline phosphatase-conjugated goat anti-mouse IgG subclass-specific Ab (Caltag Laboratories, San Francisco, CA). Isotype standards were used for standard curve fitting and Ig concentrations were calculated using Softmax software (Molecular Devices, Menlo Park, CA).
Anti-chromatin Abs were measured by ELISA as described (15) with minor modifications. Wells were coated with 100 µl of 1 µg/ml chicken chromatin. Washed wells were incubated in 100 µl of 1/100 mouse sera for 1 h at room temperature, washed, and incubated with alkaline phosphate-labeled goat-anti-IgG. Plates were developed with p-nitrophenyl phosphate (pNPP) substrate (Sigma-Aldrich, St. Louis, MO). OD405 was converted to units based on the standard curve produced by serial dilutions of pooled sera from MRL-lpr/lpr mice.
Anti-dsDNA Abs were assayed in two steps. First, a semiquantitative, but highly specific Crithidia luciliae assay was used at a serum dilution of 1/20 as described (14). Secondary Ab was FITC-conjugated goat anti-mouse IgG (Southern Biotechnology Associates). Second, positive sera were further assayed for IgG and IgG subclass-specific anti-dsDNA Ab by a less specific but much more quantitative ELISA as previously described (16). Samples were screened at a 1/50 dilution and the results were expressed in arbitrary units relative to a standard positive sample derived from MRL-lpr/lpr mice.
Assessment of nephritis
The development of proteinuria was assessed weekly using Albustix assay strips (Bayer, Elkhart, IN) using a scale of 0 to +4. Severe proteinuria was defined as >300 mg/dl (3+ or more) on two consecutive examination. For the assessment of renal histology, half of each kidney was fixed in 4% formaldehyde. Paraffin sections (3 µm) were stained with H&E, periodic acid Schiff (PAS), and Masson’s trichrome, and scored in a blinded fashion for the following features (using a 0 to 3 scale): glomerular hypercellularity, necrotizing lesions, karyorrhexis, cellular crescents and hyaline deposits (these features indicate glomerular activity score); interstitial inflammation, tubular cell necrosis and epithelial cells or macrophages in tubular lumens (tubulointerstitial activity score); glomerulosclerosis, glomerular scars, fibrous crescents, tubular atrophy, and interstitial fibrosis (chronic lesion score). The scores for individual features were summed to obtain the three main scores (glomerular activity score, tubulointerstitial activity score, and chronic lesion score) and then the three main scores were summed to obtain a composite kidney biopsy score (KBS). In addition, lupus nephritis was classified and graded histopathologically according to the World Health Organization classification and scored according to the morphologic index of glomerular activity, the method widely used in evaluation of kidney biopsies in humans with SLE.
For immunofluorescence, kidney tissue was embedded in OCT compound and unfixed frozen sections were stained with 1/20 goat anti-mouse IgG1, 2a, 2b, or 3 or IgM (Southern Biotechnology Associates) or with rabbit anti-mouse C3 (Cappel Laboratories, Durham, NC). Glomerular staining was graded by intensity (0, no staining; 4+, maximum staining) and pattern (predominantly mesangial vs capillary plus mesangial).
Statistical analyses
The data are presented as mean ± SEM. The Mann-Whitney U test was used to analyze nonparametric data for statistical significance. Values of p 
0.05 were considered significant.
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Results |
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We have transferred the Stat4 and Stat6 null mutations onto the lupus background of NZB x NZW. However, rather then using the original parental strains, we used the (NZB x NZW)F1-derived inbred mouse strains NZM 2328 which most closely resembled the (NZB x NZW)F1. This eliminated the need to backcross the null mice separately to NZB and NZW background.
A marker-assisted selection protocol was used, which accelerates the introgression of genes by using genetic markers to facilitate this process (17, 18, 19). By specifically selecting progeny at each backcross generation that carry the target gene and have the lowest content of heterozygosity throughout the genome, the rate of donor genomic segment elimination is significantly accelerated. We screened each generation with microsatellite markers at a density of 25 cM. At least 25 males were screened each generation. In all, six backcrosses were completed in each of the lines. The final backcross mice carried 76 of 76 (100%) NZM 2328-specific markers but none (0%) of the C57BL/6 or 129 strain-specific marker alleles. These mice were intercrossed to produce homozygous null mice used for all the experiments described below. To follow the Stat4 and Stat6 null mutations in the mice, we used PCR-based assays: one that amplified the neomycin-resistance (neo) gene and the other that amplified a part of the respective Stat gene containing the eliminated portion. The defect in the resulting mice was conferred by the absence of these Stat proteins in Western blot analysis using polyclonal anti-Stat4 and anti-Stat6 (data not shown).
Characterization of the mice lines produced
The first step in characterizing these newly created mice models was to verify the effect of Stat-4 and Stat-6 null mutations on the production and regulation of Th1 and Th2 cytokines.
Spleen cells from "wild-type" NZM 2328 (control) and mutant NZM.Stat4-/- and NZM.Stat6-/- mice were activated in vitro with plate-bound anti-CD3 in the absence of exogenous cytokines to determine their developmental potential into Th1 or Th2, as described (20, 21). IFN- and IL-4 production was quantified by ELISA. Representative results are shown in Fig. 1. Under neutral activation conditions, splenocytes from NZM 2328 secreted both IFN- and IL-4. Consistent with the results obtained in the original 129/B6 Stat4-/- mice, NZM 2328.Stat4-/- mice had significantly reduced levels of IFN- secretion (p < 0.01), whereas NZM 2328.Stat6-/- mice secreted high levels of IFN- but very little IL-4 (p < 0.05).
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in response to IL-12. Similarly to the original 129/B6 Stat-4-/- mice, the NZM2328.Stat4-/- mice do not show enhanced IFN- secretion following IL-12 stimulation. In contrast, NZM 2328.Stat6 null mice do demonstrate enhanced IL-12-induced IFN- secretion (Fig. 2).
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Effects of Stat4 and Stat6 KO on autoimmune disease development in the NZM mouse lines
In comparison to wild-type NZM 2328 Stat4-sufficient mice, NZM 2328 Stat4-deficient female mice develop accelerated kidney disease, as measured by proteinuria and mortality (Fig. 6). The fact that proteinuria levels closely correlated with animal mortality supports the notion that these mice die as a consequence of kidney disease. In contrast, NZM2328.Stat6-deficient female mice showed a significant delay in development of kidney disease (p < 0.001).
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For assessment of renal pathology, paraffin sections were stained with H&E, PAS, and Masson’s trichrome, and scored in a blinded fashion. The severity of kidney pathology was graded according to the World Health Organization classification followed by computing a composite KBS for each kidney evaluated. By the age of 5.5 mo, most Stat4-deficient female mice showed diffuse proliferative GN World Health Organization class IV or diffuse proliferative and sclerosing GN class IV (Fig. 8). Stat 6-deficient female mice showed very little pathology at 5–5.5 mo of age. By 10–12 mo of age, they showed mostly mesangioproliferative GN class II or focal proliferative GN class II. As shown in Fig. 8F, by 5.5 mo of age Stat4-/-.NZM 2328 showed significant increase in the composite KBS compared with wild-type NZM 2328 mice (p < 0.05). In contrast, Stat6-deficient NZM 2328 mice had a significant decrease in KBS at 5.5 mo of age (p < 0.01), and even at 10–12 mo of age (p = 0.05), an age when all Stat4-deficient mice and most control NZM 2328 mice were already dead. The Stat4 null mice could not be differentiated from Stat6 null mice by the extent of involvement of interstitium and blood vessels or on the level of evidence for glomerulosclerosis vs glomerular infiltration (data not shown).
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Discussion |
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The Stat4 and Stat6 genes encode transcription factors that, when phosphorylated by Janus kinases, are activated and transported to the nucleus where they regulate cytokine-induced gene expression (22). Until very recently, production of IL-4, an essential cytokine for differentiation of Th2-responsive cells, was thought to be completely dependent on signaling via Stat6. Indeed, original studies with Stat6-/- mice suggested that these KO mice were completely deficient in Th2 production (20, 21, 23). However, more recent studies indicate there is an alternative pathway for differentiation of Th2 cells that is independent of Stat6 (24, 25, 26, 27). Despite this alternative pathway, CD4+ T cell differentiation in Stat6-/- mice defaults to a Th1-like response and a deficiency of functional Th2 cells (26, 28, 29).
Similarly, Stat4 was originally thought to be critical for development of Th1 cells (30) because it is a transcriptional regulatory molecule for IL-12, which is a potent differentiation agent for Th1 cells (31). However, recent studies demonstrate that there are both Stat4-dependent and Stat4-independent pathways for the development of Th1 cells. Although the Stat4-dependent pathway involves induction of IL-12, the Stat4-independent pathway does not, and thereby represents a default pathway that occurs in the absence of Stat4 signaling (32). Even though Stat4-/- mice contain an alternative pathway for generating Th1 cells, the CD4+ T cell response in these mice is strongly skewed toward a Th2 response, and Stat4-/- mice are deficient in Th1-type cells.
There is great interest in the possibility that a skewing toward a Th2-type response plays a major role in murine lupus. Such skewing has been suggested because CD4+ T lymphocytes from (NZB x NZW)F1 mice secrete less and less IL-2 as the mice age and the disease becomes more clinically evident (33). Further, IL-4 mRNA expression increases with age and disease development in (NZB x NZW)F1, while IL-12 mRNA expression decreases (34). Cytokine profiles in mitogen-stimulated T cells in these mice showed high Th2-type cytokine production (IL-4, IL-5, and IL-10), while Th1 cytokine (IL-2, IFN-) production appears to be low (10). Moreover, the administration of anti-IL-10 to (NZB x NZW)F1 mice delays disease onset (35), while treatment with IL-4 antagonists decreases anti-DNA Ab and associated renal disease (36). Also, transgenic expression of IL-4 under a MHC class I promoter induces autoimmunity in a nonautoimmune background (37).
Although the above studies suggest a pivotal role for Th2 cytokines, other findings suggest an important role for Th1 responses, as well. For example, the major isotype eluted from glomerular lesions of lupus mice is IgG2a (38), which depend on IFN- for its synthesis. Furthermore, lupus mice are protected from disease development by inhibition of IFN- signaling. Thus, it has been shown that neutralization of IFN- by mAb (8) or soluble IFN- receptor (9) prevent GN in (NZB x NZW)F1 mice, while administration of exogenous IFN- accelerates renal disease. Interestingly, deficiency in IFN- receptor crossed onto NZB and NZW mice causes a marked reduction in anti-DNA Ab, including IgM and IgG1 as well as IgG2a (39). Further, IFN- receptor null (NZB x NZW)F1 mice show significant reduction in kidney disease development (39).
Our results presented in this study suggest that the validity of the Th1/Th2 dichotomy in explaining the pathogenesis of lupus appears too simplistic. Some recent studies have raised doubts about the validity of the Th1/Th2 paradigm in other systems as well (40, 41).
The results raise questions regarding the accepted paradigm that autoantibodies (especially anti-dsDNA Ab) play a major, direct, and culpable role in inducing some of the disease manifestations of SLE (especially nephritis). It is commonly believed that if Ab-Ag interaction occurs and complement is activated, tissue damage will result. The mechanism by which anti-DNA Ab is believed to fix to tissue and cause damage is either by forming immune complexes of the correct size and charge to be trapped in the basement membranes of glomeruli, or by binding directly to DNA, DNA-histone, or to non-DNA Ag present in the basement membranes. Our results suggest that the association between the presence of autoantibody and kidney disease is not a simple and direct cause-effect relationship–any explanation reduced to such a simple cause-effect relationship is precarious, at best.
Recently, Shlomchik and colleagues (42) have demonstrated an Ab-independent mechanism for renal and vascular disease in the MRL-lpr/lpr mouse model using a mutant transgene encoding membrane Ig, but which did not permit the secretion of circulating Ab. Thus, a possible explanation of the accelerated nephritis obtained in Stat4-deficient lupus mice might be that autoantibodies are not critical for the development of lupus nephritis. However, while the membrane only Ig transgenic MRL-lpr/lpr mice showed preferential involvement of the interstitium and blood vessels rather than proliferative GN, there seems to be no such preferential involvement in the Stat4-deficient NZM2328 mice. In contrast, the genetic background of the mice developing the Ab-independent lupus nephritis is very different from the background of the Stat 4 null mice. Thus, the differences in the nature of the nephritis between these mice might be a consequence of their different genetic background. An alternative mechanistic explanation might be that for developing lupus nephritis a certain threshold of insult to the kidney is necessary to initiate the process. Presumably, such an insult can be delivered by autoantibody complement complexes. If the threshold of the insult is achieved, the additional excess of autoantibody might make no difference in terms of the overall outcome of the kidney disease. We are currently testing these possibilities.
Our results in Stat4-deficient NZM 2328 mice also substantiate the importance of Th1-type cytokines in autoantibody production. These results also raise questions regarding the claimed crucial role of IFN- in the development of lupus pathology because we found that the nephritis in Stat4 null mice is worse than in wild-type NZM 2328 mice despite reduced production of IFN-. Furthermore, the data might suggest a protective role for IFN- in disease initiation, possibly by a mechanism similar to what has been shown in transplantation studies (43, 44) and in some cases of autoimmunity (45, 46, 47). Further studies will attempt to verify these possibilities.
It is also possible that the observed effects of the Stat4 or Stat6 KO reflect the removal of lupus susceptibility or resistance genes during backcrossing of the mutated locus from the 129/C57BL6 background on to the NZM.2328 genetic background. Stat4 and Stat6 genes, however, are on mouse chromosome 1 (25.9 cM) and mouse chromosome 10 (70.0 cM), respectively; thus, both genes are clearly outside any known lupus susceptibility or resistance region in NZM 2328 mice (5). Therefore, it is unlikely that the phenotypes presented by these mice are due to any interference of the mutated locus with potential lupus susceptibility genes.
More importantly, these findings may be relevant to human SLE because there is abundant clinical experience demonstrating that severe GN can arise even in the absence of elevated anti-dsDNA Ab. In addition, in our lupus clinic we are following several patients that have stable high titers of anti-dsDNA Ab in their serum, without any evidence of kidney disease and a very mild SLE, an occurrence that is reminiscent of our Stat6-deficient NZM 2328 mice.
Finally, our results suggest that development of drugs that specifically regulate Stat4 and Stat6 activity may pave the way for novel therapeutic intervention in SLE.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Chaim O. Jacob, Department of Medicine, University of Southern California School of Medicine, 2011 Zonal Avenue, HMR 705, Los Angeles, CA 90089. E-mail address: Jacob{at}usc.edu
3 Abbreviations used in this paper: SLE, systemic lupus erythematosus; KO, knockout; GN, glomerulonephritis; NZM, New Zealand mixed; PAS, periodic acid Schiff; KBS, kidney biopsy score.
Received for publication February 21, 2003. Accepted for publication May 27, 2003.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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. J. Exp. Med. 166:798. receptor. Eur. J. Immunol. 25:6.[Medline]
:interleukin-4 secreting cells in MRL-lpr/lpr mice. Autoimmunity 21:107.[Medline]
is required for lupus nephritis in mice treated with the hydrocarbon oil pristine. Kidney Int. 60:2173.[Medline]
receptor deletion prevents autoantibody production and glomerulonephritis in lupus-prone (NZB x NZW)F1 mice. J. Immunol. 160:3713. and IL-4 in acute graft-vs-host disease after allogeneic bone marrow transplantation in mice. J. Clin. Invest. 102:1742.[Medline]
alters the pathology of graft rejection: protection from early necrosis. J. Immunol. 166:7072.-interferon in DP-BB rats. Diabetes 47:32.[Abstract]
. Eur. J. Immunol. 26:2393.[Medline]
axis in the development of autoimmune myocarditis: induction by IL-12 and protection by IFN-. J. Immunol. 167:5464.This article has been cited by other articles:
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  H. Agrawal, N. Jacob, E. Carreras, S. Bajana, C. Putterman, S. Turner, B. Neas, A. Mathian, M. N. Koss, W. Stohl, et al. Deficiency of Type I IFN Receptor in Lupus-Prone New Zealand Mixed 2328 Mice Decreases Dendritic Cell Numbers and Activation and Protects from Disease J. Immunol., November 1, 2009; 183(9): 6021 - 6029. [Abstract] [Full Text] [PDF]
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 A-K Abelson, A M Delgado-Vega, S V Kozyrev, E Sanchez, R Velazquez-Cruz, N Eriksson, J Wojcik, M V P Linga Reddy, G Lima, S D'Alfonso, et al. STAT4 associates with systemic lupus erythematosus through two independent effects that correlate with gene expression and act additively with IRF5 to increase risk Ann Rheum Dis, November 1, 2009; 68(11): 1746 - 1753. [Abstract] [Full Text] [PDF]
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N. Jacob, H. Yang, L. Pricop, Y. Liu, X. Gao, S. G. Zheng, J. Wang, H.-X. Gao, C. Putterman, M. N. Koss, et al. Accelerated Pathological and Clinical Nephritis in Systemic Lupus Erythematosus-Prone New Zealand Mixed 2328 Mice Doubly Deficient in TNF Receptor 1 and TNF Receptor 2 via a Th17-Associated Pathway J. Immunol., February 15, 2009; 182(4): 2532 - 2541. [Abstract] [Full Text] [PDF]
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 P. V. Kirjavainen, S. Pautler, M. L. Baroja, K. Anukam, K. Crowley, K. Carter, and G. Reid Abnormal Immunological Profile and Vaginal Microbiota in Women Prone to Urinary Tract Infections Clin. Vaccine Immunol., January 1, 2009; 16(1): 29 - 36. [Abstract] [Full Text] [PDF]
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|
 S. Sigurdsson, G. Nordmark, S. Garnier, E. Grundberg, T. Kwan, O. Nilsson, M.-L. Eloranta, I. Gunnarsson, E. Svenungsson, G. Sturfelt, et al. A risk haplotype of STAT4 for systemic lupus erythematosus is over-expressed, correlates with anti-dsDNA and shows additive effects with two risk alleles of IRF5 Hum. Mol. Genet., September 15, 2008; 17(18): 2868 - 2876. [Abstract] [Full Text] [PDF]
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 E. F. Remmers, R. M. Plenge, A. T. Lee, R. R. Graham, G. Hom, T. W. Behrens, P. I.W. de Bakker, J. M. Le, H.-S. Lee, F. Batliwalla, et al. STAT4 and the Risk of Rheumatoid Arthritis and Systemic Lupus Erythematosus N. Engl. J. Med., September 6, 2007; 357(10): 977 - 986. [Abstract] [Full Text] [PDF]
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 D. A. Isenberg, J. J. Manson, M. R. Ehrenstein, and A. Rahman Fifty years of anti-ds DNA antibodies: are we approaching journey's end? Rheumatology, July 1, 2007; 46(7): 1052 - 1056. [Abstract] [Full Text] [PDF]
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V. Sanchez-Guajardo, C. Tanchot, J. T. O'Malley, M. H. Kaplan, S. Garcia, and A. A. Freitas Agonist-Driven Development of CD4+CD25+Foxp3+ Regulatory T Cells Requires a Second Signal Mediated by Stat6 J. Immunol., June 15, 2007; 178(12): 7550 - 7556. [Abstract] [Full Text] [PDF]
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C. O. Jacob, L. Pricop, C. Putterman, M. N. Koss, Y. Liu, M. Kollaros, S. A. Bixler, C. M. Ambrose, M. L. Scott, and W. Stohl Paucity of Clinical Disease despite Serological Autoimmunity and Kidney Pathology in Lupus-Prone New Zealand Mixed 2328 Mice Deficient in BAFF J. Immunol., August 15, 2006; 177(4): 2671 - 2680. [Abstract] [Full Text] [PDF]
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 G. Rajagopalan, K. Iijima, M. Singh, H. Kita, R. Patel, and C. S. David Intranasal Exposure to Bacterial Superantigens Induces Airway Inflammation in HLA Class II Transgenic Mice Infect. Immun., February 1, 2006; 74(2): 1284 - 1296. [Abstract] [Full Text] [PDF]
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 K. J. Land, P. Gudapati, M. H. Kaplan, and G. S. Seetharamaiah Differential Requirement of Signal Transducer and Activator of Transcription-4 (Stat4) and Stat6 in a Thyrotropin Receptor-289-Adenovirus-Induced Model of Graves' Hyperthyroidism Endocrinology, January 1, 2006; 147(1): 111 - 119. [Abstract] [Full Text] [PDF]
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H.-J. Shin, H.-Y. Park, S.-J. Jeong, H.-W. Park, Y.-K. Kim, S.-H. Cho, Y.-Y. Kim, M.-L. Cho, H.-Y. Kim, K.-U. Min, et al. STAT4 Expression in Human T Cells Is Regulated by DNA Methylation but Not by Promoter Polymorphism J. Immunol., December 1, 2005; 175(11): 7143 - 7150. [Abstract] [Full Text] [PDF]
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A. Mathian, A. Weinberg, M. Gallegos, J. Banchereau, and S. Koutouzov IFN-{alpha} Induces Early Lethal Lupus in Preautoimmune (New Zealand Black x New Zealand White)F1 but Not in BALB/c Mice J. Immunol., March 1, 2005; 174(5): 2499 - 2506. [Abstract] [Full Text] [PDF]
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S. Lawman, C. Mauri, E. C. Jury, H. T. Cook, and M. R. Ehrenstein Atorvastatin Inhibits Autoreactive B Cell Activation and Delays Lupus Development in New Zealand Black/White F1 Mice J. Immunol., December 15, 2004; 173(12): 7641 - 7646. [Abstract] [Full Text] [PDF]
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 O Y Jones, P J Alexander, A Lacson, F Gok, A Feliz, Y Marikar, C Madivi, J M Jones, and R A Good Effects of fludarabine treatment on murine lupus nephritis Lupus, December 1, 2004; 13(12): 912 - 916. [Abstract] [PDF]
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K. J. Land, J. S. Moll, M. H. Kaplan, and G. S. Seetharamaiah Signal Transducer and Activator of Transcription (Stat)-6-Dependent, But Not Stat4-Dependent, Immunity Is Required for the Development of Autoimmunity in Graves' Hyperthyroidism Endocrinology, August 1, 2004; 145(8): 3724 - 3730. [Abstract] [Full Text] [PDF]
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 S. T. Waters, M. McDuffie, H. Bagavant, U. S. Deshmukh, F. Gaskin, C. Jiang, K. S.K. Tung, and S. M. Fu Breaking Tolerance to Double Stranded DNA, Nucleosome, and Other Nuclear Antigens Is Not Required for the Pathogenesis of Lupus Glomerulonephritis J. Exp. Med., January 20, 2004; 199(2): 255 - 264. [Abstract] [Full Text] [PDF]
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300 mg/dl) proteinuria and as percent cumulative survival at the ages shown. *, p < 0.001 vs control mice.
, Stat4-/-; 
, Stat6-/-. The mean Ab level of 8–12 mice per group at the indicated ages is shown. A, Total IgG anti-chromatin Ab levels shown in the different groups. B, Total IgG anti-dsDNA Ab levels in the mice groups and (C) serum IgG1 and IgG2a levels of anti-dsDNA Ab in the NZM2328 mice with the indicated genotypes.
