Genetic analysis of the imperfect association of H-2 haplotype with lupus-like autoimmune disease.

Abstract

Proteinuria, an indicator of renal disease, was evaluated monthly by using tetrabromphenol paper, and was graded 0-3+. Trace proteinuria is <30 mg/dl, 1+ detects 30 mg/dl, 2+ detects 100 mg/dl, and 3+ detects >500 mg/dl. Severe proteinuria was defined as >2+, and animals with this level of proteinuria at 11 months of age or earlier were classified as having severe renal disease (2, 8). These mice usually died from renal failure within 4-8 weeks after development of severe proteinuria. Animals with negative or trace proteinuria at 12 months of age were classified as having no evidence of renal disease.Southern Analysis of Genomic DNA. Liver and kidney DNA from each animal was isolated as described (9). DNA (10 pkg) was digested with restriction enzymes at a concentration of 2 units per ,ug of DNA for 1 hr at 370C. This procedure was repeated once. Various enzymes were utilized depending on the probe used: Pvu II for I-Aa, BamHI for C3, Pvu II for TU66, and Bgl II for both TU169 and the a-globin pseudogene 4 (a-q4). Digested DNAs were subjected to electrophoresis through agarose gels (10). A 0.7% gel was used for MHC, TU66, TU169, and a-+4, and the gels were run for 20 hr. To separate the fragments hybridizing to the C3 probe, a 1.5% gel was run for 48 hr. The DNA was transferred to nitrocellulose filters, baked, and hybridized with a 32p labeled probe as described (10), except that the depurination step was omitted and the gel was irradiated for 7 min with shortwave UV light before denaturation to facilitate transfer ofDNA to nitrocellulose (11). Hybridized filters were washed in 2x SSC/1% SDS at room temperature followed by 0.1X SSC/1% SDS at 550C for 45 min (lx SSC = 0.15 M Abbreviation: MHC, major histocompatibility complex.fTo whom reprint requests should be addressed. 7552The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

Induction of cross-reactive anti-dsDNA antibodies in preautoimmune NZB/NZW mice by immunization with bacterial DNA.

“…Indeed, studies of NZB/NZW as well as other lupus strains indicate defects in tolerance induction, B cell responsiveness, and patterns of V gene and idiotype expression (19)(20)(21). We have suggested therefore that the preimmune repertoire of lupus mice is uniquely enriched in autoreactive B cells that can be mutated to high affinity anti-dsDNA when appropriately stimulated.…”

“…To test this possibility, we have determined the anti-dsDNA response of young NZB/NZW female mice immunized with dsDNA from Escherichia coli (EC). These Fl hybrids spontaneously produce anti-DNA as they age and have been extensively studied as a model of lupus (19)(20)(21) In the third immunization experiment, mice were prebled and then bled weekly until one week following the last immunization.…”

Epistatic Suppression of Systemic Lupus Erythematosus: Fine Mapping ofSles1to Less Than 1 Mb

“…Earlier studies using the classic (NZB ϫ NZW]F 1 (BWF 1 )) model, from which NZM2410 is derived, in various backcross analyses have suggested that the NZW allele at the H2 (H2 z ) confers a high risk of developing renal disease, and in particular, it is the NZW class II alleles that are the dominant contributors to disease in these models (39). Many of these studies have documented that heterozygosity at the H2 gene complex is crucial for lupus susceptibility and is associated with many of the phenotypic characteristics of lupus, such as GN development, autoantibody production, and splenomegaly (40).…”

A Novel Susceptibility Locus On Chromosome 2 in the (New Zealand Black × New Zealand White)F1 Hybrid Mouse Model of Systemic Lupus Erythematosus

“…T wo of the most common features in both human and mouse models of systemic lupus erythematosus (SLE) 3 are: 1) Abs against various nuclear constituents (ssDNA, dsDNA, histone, and chromatin); and, 2) glomerulonephritis (GN) from the deposition of immune complexes onto the glomerulus basement membrane in the kidney. Even though the etiology and the specific contributions of particular gene(s) to this disease remain unsolved, substantial evidence from multiple studies in murine and human SLE has suggested that SLE has a strong genetic basis and is a complex genetic trait.…”