The JI
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     
 

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jordan, M. A.
Right arrow Articles by Baxter, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jordan, M. A.
Right arrow Articles by Baxter, A. G.
Right arrowPubmed/NCBI databases
*Gene
*Substance via MeSH
Medline Plus Health Information
*Diabetes Type 1
*Lupus
The Journal of Immunology, 2000, 165: 1673-1684.
Copyright © 2000 by The American Association of Immunologists

Linkage Analysis of Systemic Lupus Erythematosus Induced in Diabetes-Prone Nonobese Diabetic Mice by Mycobacterium bovis1

Margaret A. Jordan2, Pablo A. Silveira2, Darren P. Shepherd, Clara Chu, Simon J. Kinder, Jianhe Chen, Linda J. Palmisano, Lynn D. Poulton and Alan G. Baxter3

Centenary Institute of Cancer Medicine and Cell Biology, Newtown, Australia


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Systemic lupus erythematosus induced by Mycobacterium bovis in diabetes-prone nonobese diabetic mice was mapped in a backcross to the BALB/c strain. The subphenotypes—hemolytic anemia, antinuclear autoantibodies, and glomerular immune complex deposition—did not cosegregate, and linkage analysis for each trait was performed independently. Hemolytic anemia mapped to two loci: Bah1 at the MHC on chromosome 17 and Bah2 on distal chromosome 16. Antinuclear autoantibodies mapped to three loci: Bana1 at the MHC on chromosome 17, Bana2 on chromosome 10, and Bana3 on distal chromosome 1. Glomerular immune complex deposition did not show significant linkage to any genomic region. Mapping of autoantibodies (Coombs’ or antinuclear autoantibodies) identified two loci: Babs1 at the MHC and Babs2 on distal chromosome 1. It has previously been reported that genes conferring susceptibility to different autoimmune diseases map nonrandomly to defined regions of the genome. One possible explanation for this clustering is that some alleles at loci within these regions confer susceptibility to multiple autoimmune diseases—the "common gene" hypothesis. With the exception of the H2, this study failed to provide direct support for the common gene hypothesis, because the loci identified as conferring susceptibility to systemic lupus erythematosus did not colocalize with those previously implicated in diabetes. However, three of the four regions identified had been previously implicated in other autoimmune diseases.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Systemic lupus erythematosus (SLE)4 is a chronic, potentially fatal autoimmune disease characterized by unpredictable exacerbations and remissions of clinical manifestations involving the joints, skin, kidney, brain, serosa, lung, heart, and gastrointestinal tract. The prevalence of SLE in Caucasian populations is around 0.2% (1, 2), but a telephone survey commissioned in 1995 by the Lupus Foundation of America suggested a prevalence in the United States as high as 0.6% (3).

The cause(s) of SLE are unknown, but environmental and genetic factors are involved. The environmental factors that may trigger the disease include infections, antibiotics (especially those in the sulfonamide and penicillin groups), and UV light. SLE is known to cluster within families, and the risk of a first-degree relative of a patient developing disease is about 4% (i.e., the {lambda}s is about 20) (4). As the disease exhibits complex genetics and significant heterogeneity in terms of the clinical features at presentation, the introduction of animal models, principally mouse models, has generated considerable interest.

We have previously reported a novel model of SLE in which a single dose of pasteurized Mycobacterium bovis administered i.v. to prediabetic nonobese diabetic (NOD) mice prevented the spontaneous onset of type 1 (autoimmune) diabetes but precipitated a systemic "autoimmune rheumatic disease" (ARD) similar to SLE (5, 6). This syndrome was characterized by hemolytic anemia (HA), antinuclear autoantibodies (ANA), increased severity of sialadenitis, and glomerular immune complex deposition. The specificity of the ANA responses in these mice was directed against dsDNA and the Smith (Sm)/ribonucleoprotein complex, of which the 28-kDa polypeptide appeared to be immunodominant (7). The IgG subclass involved in the anti-Sm response was primarily IgG2a, while the subclass of the response against dsDNA was mixed, with IgG2a and IgG2b being present in equal amounts. The anti-dsDNA and anti-Sm reactivities were not mediated by polyreactive Abs because neither Ag could cross-compete plasma Ab binding to the other in competitive ELISA. The role of polyclonal B cell activation was examined by measuring total {gamma}-globulin as well as IgG reactive with other nuclear Ags including Ro60, Ro52, and La, which although not a major component of the autoantibody responses in these mice did show small but significant increases following immunization with M. bovis. Thus, polyclonal stimulation, while likely to be occurring, was not directly responsible for production of anti-Sm Abs (7). The pattern of renal disease seen in NOD mice treated with bacillus Calmette-Guérin (BCG) is similar to mild focal lupus nephritis and is characterized by segmental proliferation of a minority of glomeruli and widespread C3c deposition.

This model of SLE has two major advantages. First, it is the only mouse model for which the environmental trigger has been identified. Therefore, it introduces the opportunity for a detailed analysis of environment/genome interactions in determining disease liability. The second advantage is that the mouse strain involved, the NOD mouse strain, has been extensively genetically characterized, which permits the direct comparison of genetic loci identified in linkage studies of SLE in this model with loci previously identified in linkage analysis of type 1 diabetes.

The coexistence of these two distinct autoimmune diseases within the same mouse strain, and the reciprocal switching between these two phenotypes by a single environmental trigger (mycobacterial exposure), raises the possibility that genetic susceptibility for diabetes and SLE may be conferred by a single collection of genes. That is, a single inherited disease (autoimmunity) can be expressed as either phenotype, depending on, for example, environmental modifiers. Recent genomic studies of autoimmune diseases, both in humans and in mice, have provided significant support for this "common gene" hypothesis (8, 9, 10, 11). The linkage analysis reported here allows comparison with the data obtained from mapping diabetes in the same strain and SLE in other mouse models.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Mice

NOD/Lt, BALB/c, and C57BL/6J mice were obtained from the Animal Resource Center (Canning Vale, Australia). Breeding of specific crosses was performed within the animal facility at the Centenary Institute (Sydney, Australia). Mice were housed in clean conditions, and sentinel mice were tested by serology at 4-mo intervals for the following pathogens: mouse hepatitis virus, rotavirus, ectomelia, mouse CMV, polyoma virus, murine adenovirus, lymphocytic choriomeningitis virus, mouse pneumonia virus, reovirus, Sendai virus, Theiler’s murine encephalitis virus, Bacillus piliformis, Mycoplasma pulmonis, Bordetella bronchiseptica, Corynebacterium kutscheri, Klebsiella species, Pasteurella multocide, Pasteurella pneumotropica, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae, Citrobacter freundii, and Salmonella species. No sentinel mice tested positive for any of these pathogens.

M. bovis injection

Mice were injected at 8 wk of age with 0.4 mg of CSL (Melbourne, Australia) dried BCG (M. bovis) vaccine (living) for percutaneous use after pasteurization at 65°C for 45 min. In vitro culture of this quantity of BCG indicated that it was equivalent to ~4 x 106 CFU before inactivation.

Phenotyping

Assessment of anemia. Mice were bled 6 mo after BCG injection by retroorbital venepuncture. Hematocrits were determined by centrifuging the blood in heparinized capillary tubes (Becton Dickinson, Parsippany, NJ) at 1000 x g for 30 min. Direct Coombs’ tests were performed by washing the packed red cells three times in PBS plus 0.3% BSA and incubating them in round-bottom microtiter plates with anti-IgG serum (Southern Biotechnology Associates, Birmingham, AL) for 2 h at 37°C. Plates were then assessed visually for evidence of agglutination.

Assessment of antinuclear autoantibodies. Sera were diluted to 1:10 in PBS and incubated on HEp-2 slide monolayers (Quantafluor, Chaska, MN). Slides were washed in PBS and incubated with 20 µg/ml human-adsorbed anti-IgG FITC-conjugated antiserum (Southern Biotechnology Associates). After washing, the presence of ANA was detected by fluorescence microscopy. Samples that contained ANA were then retested at 1:100. Sera from affected NOD mice and unaffected BALB/c mice were used as positive and negative controls, respectively. Samples were declared positive for ANA if they showed moderate to strong immunofluorescent staining of HEp-2 cell nuclei at a dilution of 1:100. Samples were declared negative for ANA if they showed no evidence of immunofluorescent staining of HEp-2 cell nuclei at a dilution of 1:10.

Detection of glomerular immune complex deposition. Unlike that in NZB/W mice, the pattern of renal disease in NOD mice treated with BCG is similar to mild focal lupus nephritis and is characterized by widespread C3c deposition and segmental proliferation of a minority of glomeruli. As this form of nephritis is associated with only mild proteinuria, measurement of urinary protein with clinical dip-sticks was found not to be a robust diagnostic test. Although the diagnosis can be readily made by electron microscopy, the very large numbers of samples in this study precluded this option. In view of the relative subtlety of the histological changes as well as the expected increase in variation in the backcross compared with examining genetically identical parental or F1 mice, glomerular C3c deposition was used to detect the presence of renal disease because it was the most robust assay available.

Mice were killed at 32 wk, and their tissues were harvested for histology and DNA extraction. Kidneys were snap frozen, and C3c glomerular deposition was detected on cryostat sections following incubation with 1:100 dilution of FITC-conjugated goat anti-mouse C3c serum (Nordic Immunology, Tilburg, The Netherlands) in PBS for 30 min at room temperature. Slides were then washed three times for 5 min with PBS, mounted, and examined by fluorescence microscopy.

Genotyping

DNA of BC1 progeny was extracted from livers and subjected to an autosomal genome-wide scan at a 15-centiMorgan (cM) resolution using 150 microsatellite markers polymorphic between BALB/c and NOD/Lt strains chosen from the Whitehead Institute simple sequence length polymorphism library (12). A list of all microsatellite markers typed is available on request. Analysis of simple sequence length polymorphism was performed as previously described (13).

Linkage analysis

Genotyping errors were identified manually as double recombinants or by the error checking function of Mapmaker/EXP (14) and were reamplified. Recombination distances between markers were calculated from recombination frequencies using the Mapmaker/EXP program (14). Lengths of chromosomes and order of markers were checked against published maps (15) (http://www-genome.wi.mit.edu/; http://www.informatics.jax.org/). Two approaches to linkage analysis were used. To calculate the linkage of individual markers to the autoimmune phenotypes, a 2 x 2 contingency table ({chi}2 test of independence) was applied. The significance of linkage achieved by this method was tested by creating experimentwise thresholds using permutation analysis (16). The thresholds were simultaneously valid for all markers of the genome and were calculated by permuting the phenotype data 10,000 times. The highest {chi}2 value throughout the whole genome for each permutation was collected and subsequently sorted to produce a null distribution of the test statistic. The distribution allowed the calculation of experimentwise 95% (p < 0.05 of type 1 error in the entire genome) and 37% (p < 0.63 of type 1 error throughout the genome) critical thresholds. Markers with experimental {chi}2 surpassing the experimentwise 95% critical threshold were considered significantly linked to the trait, while those surpassing the 37% threshold were considered to show a trend toward linkage (i.e., suggestive linkage) to the trait.

In an alternative approach, interval analysis of linkage to the autoimmune phenotypes was conducted on two linkage analysis programs. The MapManager/QT program (17) was initially used to perform interval linkage analysis on the autoimmune phenotypes by entering the trait values as 0 for unaffected and 1 for affected mice (Ref 18 ; Kenneth Manly, unpublished observations). The degree of linkage to the autoimmune phenotypes was reported using a likelihood ratio statistic (LRS) (19). The program was used to calculate permutation-derived threshold values for significant and suggestive linkage (according to Lander and Kruglyak’s specifications; Ref. 20) for the interval analysis, utilizing the methods established by Churchill and Doerge (16, 17, 20). One thousand permutations of phenotypes in the dataset were performed. Qualitative trait analysis was also performed on Mapmaker/QTL 2.0b using the "penetrance scan" function (Ref. 21 ; new version supplied by Mark Daly of the Whitehead Institute of Biomedical Research), which optimizes a set of penetrances for each genotypic class. Trait values were entered as 0 or 1 for unaffected and affected mice, respectively. Results of the penetrance scan are given as a logarithm of odds (LOD) score, representing the likelihood that data have arisen due to the effect of a qualitative trait locus with optimized sets of penetrances rather than the effect of chance (under the null hypothesis that the penetrances for all genotypic classes are equal). Significance thresholds used were those suggested by Lander and Kruglyak (20) for analysis of a mouse backcross; viz LOD >= 3.3 for the threshold for significant linkage and LOD >= 1.9 for the threshold suggestive of linkage. These values were validated by substitution of the following specific values from our genetic study into equation 1 (µ(T) = [C + 2{rho}GT2){alpha}(T)) in Lander and Kruglyak’s paper (20): C (number of chromosomes) = 19, {rho} (measure of S(x) fluctuation, which represents total crossover rate for a backcross with 1 df) = 1, G (genome size in Morgans) = 15.3 (see Results), T (pointwise significance level given in {chi}2 (1 df)) = 3.841, and {alpha}(T) = 0.05. From this equation, we expected the pointwise significance level of p = 0.05 to be surpassed approximately seven times by chance (µ(T)) in each of the linkage analyses. Thus, it was determined that a pointwise significance level of p = 0.0001 and p = 0.0036 would only be surpassed 0.05 and 1 time throughout the genome by chance, respectively. These figures correspond to LOD values of 3.3 (p = 0.0001) and 1.8 (p = 0.0036), respectively, which are similar to those suggested by Lander and Kruglyak (20). These values did not change when µ(T) was calculated separately for each trait.

Mapmaker/QTL and MapManager/QT have individual advantages, and the two programs apply slightly different methods to calculate linkage to binary traits. Mapmaker/QTL is usually regarded as the tool of choice for the analysis of murine data sets, and its "penetrance scan" function is ideally suited for performing interval linkage analysis of binary traits. One disadvantage of the program is its tendency to produce artificially elevated LOD scores in large intervals (>15 cM) (21), which greatly limits its utility for calculating significance thresholds by permutation analysis. MapManager/QT has the advantage of being able to accurately calculate permutation-derived thresholds specific for individual datasets using methods derived from Churchill and Doerge (16).

Disease genes were proposed within a region when the significance thresholds were surpassed in at least two of the three analyses performed.

Other statistical analyses

Qualitative differences between samples were examined using the 4-fold table ({chi}2) test unless the expected value in any cell was 5 or less, in which case the Fisher’s exact test was used. Quantitative differences between samples were compared using the Mann-Whitney U (rank sum) test. A goodness of fit ({chi}2) test was used to compare proportions of affected animals with those predicted by modeling.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Phenotypic analysis of parental and F1 hybrid mice

NOD/Lt, C57BL/6, BALB/c, and hybrid mice (produced by mating NOD/Lt mice with either C57BL/6 or BALB/c mice) were injected i.v. at 8 wk with 0.4 mg heat-killed M. bovis. After 6 mo, the mice were bled to detect HA (by measurement of hematocrit) and ANA (by HEp-2 immunofluorescence) and subsequently killed for removal of kidneys for immunofluorescent staining of glomerular C3c deposits.

The hematocrits of all animals tested were normally distributed about a mean of 53% with a SD of 3.8%. Therefore, anemia was defined as a drop in the hematocrit to 1.645 SD below the mean (i.e., hematocrit <47%; p < 0.05). While the great majority of female NOD/Lt mice became anemic (9 of 11; 82%), only one of 14 (7%) female C57BL/6 mice and none of 16 female BALB/c mice did so (Table IGo; p < 0.0001; 4-fold table {chi}2 test). Similarly, although the sera of 10 of 11 (91%) female NOD/Lt mice had moderate to strong immunofluorescent staining of HEp-2 cell nuclei at a dilution of 1:100, none of the sera from 14 C57BL/6 and 16 BALB/c female mice did so (Table IGo; p < 0.0001; 4-fold table {chi}2 test). The differences in C3c staining deposition were less clear cut, as nine of 10 (90%) female NOD/Lt mice showed strong glomerular staining compared with four of nine (44%) C57BL/6 and three of 10 (30%) BALB/c female mice (Table IGo; p < 0.05; 4-fold table {chi}2 test).


View this table:
[in this window]
[in a new window]
  		Table I. Phenotypic analysis of parental strains and F1 hybrids1

 
The hybrid mice were almost completely protected from the induction of HA and ANA, with only two of 57 (4%) female (NOD x B6)F1 mice developing anemia (Table IGo; p < 0.0001; 4-fold table {chi}2 test). While NOD/BALB hybrid mice were free of glomerular C3c deposition, it was present in about half of NOD/B6 hybrid mice. The presence of glomerular immune complex deposition in female NOD/B6 F1 hybrid mice showed a significant deviation from expected values when the two directions of mating were compared (Table IGo; 11 of 14 vs 3 of 11; 79% vs 27%; p < 0.05; 4-fold table {chi}2 test). These two crosses were repeated, and this difference was not confirmed (3 of 10 vs 7 of 14; 30% vs 50%; NS; 4-fold table {chi}2 test) and is therefore unlikely to be of importance. No evidence was found for any other sex-linked affects in these crosses.

The penetrances of HA, ANA, and glomerular immune complex deposition in NOD mice were higher in female mice than male mice, although this trend only reached significance for glomerular immune complex deposition (p < 0.0005, 4-fold table {chi}2 test) and was not apparent in B6 or BALB/c mice or the hybrid progeny. As the phenotypic differences were more strongly marked in the BALB/c crosses than those using C57BL/6 mice, the NOD/BALB strain combination was selected for the linkage study. As considerable difficulties were experienced with conflicts arising between male hybrid mice, only female backcross mice were selected for analysis.

Production and phenotypic analysis of backcross mice

NOD/Lt mice were crossed with NOD/BALB hybrid mice in all four possible directions. At 8 wk of age, 960 segregating BC1 female mice were injected i.v. with 0.4 mg heat-killed M. bovis and 6 mo later were bled to detect HA (by measurement of hematocrit and direct Coombs’ test) and ANA (by HEp-2 immunofluorescence) and then killed for removal of kidneys and immunofluorescent staining of glomerular C3c deposits.

The hematocrits of the BC1 progeny were not normally distributed, but formed a bimodal distribution with left-skewing of the main population (Fig. 1GoA). The threshold between the two populations occurred at 44%, which correlated well with the presence or absence of Coombs’ Abs. While 36 of 45 (80%) mice with hematocrits below 44% tested positive in the direct Coombs’ test, only 47 of 820 (6%) mice with hematocrits of 44% or above did so. For the purposes of linkage analysis, HA was declared to be present in the mice with Coombs’ Abs in their sera and hematocrits below 44%. The control sample was selected from the mice without Coombs’ Abs in their sera and hematocrits of 44% or above, with a strong bias for hematocrits around the mode of the main peak in the population distribution (Fig. 1GoA). Mice with very high hematocrits were avoided because diabetics were over-represented in this sample due to dehydration and hemoconcentration secondary to hyperglycemic diuresis. As the hematocrits of BC1 progeny were not normally distributed, they did not represent a quantitative trait suitable for quantitative trait locus (QTL) mapping. Furthermore, as the threshold between normal values and anemic values correlated well with the presence of Coombs’ Abs, a binomial linkage analysis of hematocrits as a separate trait would be unlikely to contribute information independent of that obtained from mapping HA.



View larger version (52K):
[in this window]
[in a new window]
  		FIGURE 1. Phenotypic analysis of BC1 mice. A, Histogram of hematocrits of BC1 population ({blacksquare}) and the anemic () and control () samples selected for genomic analysis of HA. B, Histogram of ANA as detected by immunofluorescent staining of HEp-2 cell nuclei at a dilution of 1:10 and 1:100 in sera of BC1 population ({blacksquare}) and the affected () and unaffected () samples selected for genomic analysis.

 
As the hematocrit threshold for BC1 mice differed from that applied to the parental and F1 mice, it is of value to consider those results in the light of this threshold. Only one of 32 male BALB/c mice (3%) had a hematocrit below 44%, and none of the C57BL/6 or F1 mice did so. While four of 11 (36%) female and one of 13 (8%) male NOD mice had hematocrits <44%, another four females (total 8 of 11, 73%) had hematocrits of 44%. Two more BALB/c males (total 3 of 32, 9%) had hematocrits of 44%, while none of the C57BL/6 or F1 mice did so. Thus, it would appear that the shape of the distribution of hematocrits in BC1 mice differs slightly from that in female NOD mice. It is not known if this difference in thresholds resulted from differences in technique (the assays on parental mice were not contemporaneous with those on the BC1 mice) or from differences in the genetic background of the mice.

ANA were detected in the sera of BC1 progeny by immunofluorescence detection of IgG bound to HEp-2 nuclei at a dilution of 1:10, and positive samples were retested at a dilution of 1:100. Mice were declared positive if their sera were positive or strongly positive at 1:100 (134 of 903; 15%) and negative if their sera tested negative at 1:10 (154 of 903; 17%; Fig. 1GoB).

Glomerular C3c deposition was detected by immunofluorescence microscopy of renal cryostat sections in 68 of 818 (8.3%) BC1 progeny.

Complete phenotypic data for HA, ANA, and glomerular immune complex deposition were available for 804 BC1 female progeny. The goodness of fit ({chi}2) test was used to compare the observed frequencies of coinheritance of more than one phenotype with those expected on the basis of the observed frequencies of each individual phenotype, assuming the null hypothesis that the traits segregated independently. The null hypothesis was rejected (p < 0.0001; {chi}2 = 19.2, 2 df), almost entirely due to greater than expected coinheritance of HA and glomerular immune complex deposition. There was no evidence of coinheritance of ANA and glomerular immune complex deposition (Fig. 2Go), consistent with other reports (22) but not with a proposed critical role of ANA in the pathogenesis of glomerular immune complex deposition (reviewed in Ref. 23 ; paradox discussed in Ref. 24).



View larger version (23K):
[in this window]
[in a new window]
  		FIGURE 2. The numbers (A) and probabilities (B) of observed and expected (in parentheses) phenotype frequencies for the 804 BC1 female progeny for which complete phenotype data were available.

 
No evidence of sex-linked effects were seen in the phenotypes of BC1 progeny.

Construction of linkage map

A genome map was created by typing 221 female BC1 progeny at each of 150 polymorphic microsatellite markers distributed throughout the autosomal genome. Recombination distances between each marker were determined using the Mapmaker/EXP program (14), and the lengths of chromosomes and the order of markers were checked against published maps (15). The total autosomal genome length (excluding centromeric and telomeric portions) obtained here was 1529 cM, compared with 1187 cM reported for the MIT map, consistent with suppression of recombination in the intraspecific cross used by Dietrich et al. (15). The gene orders obtained from this data set conformed well to those previously published, with the following exceptions: D1Mit438 mapped 0.9 cM distal to D1Mit9 (order reversed); D3Mit10 mapped 2.4 cM distal to D3Mit11 (colocalized on the MIT map); D9Mit206 colocalized with D9Mit67 (2.2 cM more distal on MIT map); D14Mit225 mapped 2.6 cM distal to D14Mit160 (order reversed); D14Mit77 mapped 4.4 cM distal to D14Mit136 (colocalized on MIT map); and D17Mit83 colocalized with D17Mit16 (1 cM more distal on MIT map).

Genotypic and linkage analysis of HA

An autosomal genome scan of 34 anemic (Coombs’ positive, hematocrit <44%) and 53 unaffected female BC1 progeny was conducted using 133 polymorphic microsatellite markers with an average marker separation of 15 cM. Linkage analysis was initially performed at each marker using a 2 x 2 contingency table, and the results were compared with the 37% (suggestive) and 95% (significant) experimentwise statistical thresholds derived from a permutation analysis of 10,000 iterations of the data as specified by Churchill and Doerge (16). Genotype frequencies differed significantly ({chi}2 > 12.3) between affected and unaffected mice at loci on chromosomes 16 and 17 and exhibited results suggestive of linkage ({chi}2 > 6.5) on distal chromosome 1 and at the most proximal marker tested on chromosome 14 (Table IIGo). While NOD alleles contributed to susceptibility in a recessive fashion on chromosomes 1 and 17, BALB alleles contributed to susceptibility on chromosomes 14 and 16.


View this table:
[in this window]
[in a new window]
  		Table II. Genetic markers exhibiting evidence of linkage to HA

 
Interval analysis was performed on the data set using MapManager/QT and the penetrance scan of Mapmaker/QTL. Due to difficulties in producing permutation thresholds for Mapmaker/QTL analyses, the stringent linkage thresholds for experimental mouse backcrosses set by Lander and Kruglyak (20) for significant linkage (LOD >= 3.3) and suggestive linkage (LOD >= 1.9) were applied. The Mapmaker/QTL analysis confirmed the results obtained using contingency tables, in that it identified regions of significant linkage to HA on chromosomes 16 and 17 (Fig. 3Go). The peak linkage on chromosome 16 occurred between D16Mit58 and D16Mit5 (LOD = 3.9). The region of significant linkage on chromosome 17 extended over about 20 cM, with a peak occurring at D17Mit83 (Hsp70–1) and D17Mit16 (LOD = 4.8), which lie within the H2. A second peak was identified between D17Mit20 and D17Mit153 (LOD = 3.8), with a minimum LOD of 3.0 between the two peaks. The linkage region on distal chromosome 1 was also identified as suggestive of linkage with Mapmaker/QTL, appearing as two small peaks, one between D1Mit108 and D1Mit36, and the other between D1Mit36 and D1Mit406. Analysis of the data set using MapManager/QTL was applied using permutation derived statistical thresholds based on 1000 iterations of the data. This analysis again confirmed linkage on distal chromosome 16 and proximal chromosome 17 (data not shown)



View larger version (21K):
[in this window]
[in a new window]
  		FIGURE 3. Mapmaker/QTL linkage analyses for HA (short dash line), ANA (solid line), and autoantibodies (long dash line) on chromosomes 1 (A), 10 (B), 16 (C), and 17 (D). The straight thin line represents the Lander and Kruglyak (20 ) threshold for significance.

 
In summary, extensive analysis of this data set identified two genomic regions as being linked to HA induced by M. bovis in mice. The first, here named Bah1 (BCG-induced autoimmune hemolysis, locus 1) was mapped to proximal chromosome 17 and may be within the H2. The second locus, here named Bah2 (BCG-induced autoimmune hemolysis, locus 2) was mapped to D16Mit5 on chromosome 16.

Genotypic and linkage analysis of ANA

An autosomal genome scan of 52 female BC1 progeny with no evidence of plasma ANA (by immunofluorescence at a dilution of 1:10) and 52 female BC1 progeny with strong plasma ANA (positive at 1:100) was conducted using 127 polymorphic microsatellite markers with an average marker separation of 15 cM. Linkage analysis was initially performed at each marker using a 2 x 2 contingency table and results compared with the 37% (suggestive) and 95% (significant) experimentwise statistical thresholds derived from a permutation analysis of 10,000 iterations of the data. Genotype frequencies differed significantly ({chi}2 >= 12.0) between affected and unaffected mice at loci on chromosomes 1, 10, and 17 (Table IIIGo). Suggestive linkage ({chi}2 >= 6.5) was also identified on chromosome 5. The NOD alleles contributed susceptibility in a recessive fashion on chromosomes 1, 5, and 17, while the BALB/c allele contributed susceptibility in a dominant fashion on chromosome 10 (Table IIIGo).


View this table:
[in this window]
[in a new window]
  		Table III. Genetic markers exhibiting evidence of linkage to the production of ANA

 
Mapmaker/QTL interval analysis confirmed that loci on chromosomes 10 and 17 were linked to ANA as they exceeded the linkage thresholds set by Lander and Kruglyak for significant linkage (LOD >= 3.3; Fig. 3Go). Regions with suggestive linkage were also identified on chromosomes 1 and 5. Linkage peaked on distal chromosome 1 at D1Mit399 (LOD = 2.8), on chromosome 5 between D5Mit239 and D5Mit65 (LOD = 3.0), and on proximal chromosome 10 between D10Mit213 and D10Mit257 (LOD = 4.9). LOD scores exceeded the Lander and Kruglyak threshold for significant linkage over most of chromosome 17, with the proximal half of the chromosome surpassing a LOD of 4.0. The maximum LOD score on this chromosome (7.0) was obtained at D17Mit83, D17Mit16, and D17Mit24, which are associated with the H2.

Analysis of the data set with MapManager/QT confirmed the regions of significant (LRS >= 12.8) linkage on chromosomes 10 and 17. This analysis also confirmed the chromosome 1 linkage identified by the {chi}2 analysis. Linkage reached significance with a sharp peak at D1Mit399 (LRS = 13.5), although two other peaks exhibiting a trend toward linkage (LRS >= 6.7) were identified, one centered on D1Mit445 (LRS = 11.8) and the other on D1Mit135 (LRS = 7.9), consistent with similar results obtained with Mapmaker/QTL.

In summary, all three analyses identified genomic regions controlling susceptibility to ANA occurring in NOD mice treated with M. bovis. The first, here named Bana1 (BCG-induced ANA, locus 1), mapped to proximal chromosome 17 and may be the H2, although the breadth of the peak is suggestive of the existence of multiple genes in this region, which can influence the development of ANA in this model. The second locus, here named Bana2 (BCG-induced ANA, locus 2), mapped to proximal chromosome 10, between D10Mit213 and D10Mit257. A third locus, here named Bana3 (BCG-induced ANA, locus 3) and mapped to D1Mit399 on distal chromosome 1, achieved significance when analyzed with the contingency table and MapManager/QT, but not when analyzed with Mapmaker/QTL.

Genotypic and linkage analysis of autoantibodies

Because of the likelihood factors shared in the etiologies of Coombs’ Abs and ANA, and the apparent colocalization of genes for both traits on chromosomes 1 and 17, an autosomal genome scan was performed comparing 94 female BC1 progeny with autoantibodies (either positive for ANA at a dilution of 1:100 or direct Coombs’ test positive) and 54 unaffected female BC1 progeny (negative for ANA at 1:10 and Coombs’ negative). Linkage analysis was initially performed at each marker using a 2 x 2 contingency table and results compared with the 37% (suggestive) and 95% (significant) experimentwise thresholds derived from a permutation analysis of 10,000 iterations of the data. Genotype frequencies differed significantly ({chi}2 >= 12.2) at loci on chromosomes 1 and 17. Loci on chromosomes 5, 10, and 16 demonstrated a trend toward linkage ({chi}2 >= 6.5). Linkage peaked on chromosome 1 at D1Mit396 with a {chi}2 of 17.4, while the H2-associated markers on chromosome 17 (D17Mit83, D17Mit16, and D17Mit24) reached a {chi}2 of 42.2. The NOD alleles contributed susceptibility in a recessive fashion on chromosomes 1, 5, and 17, while the BALB/c alleles contributed to susceptibility in a dominant fashion on chromosomes 10 and 16 (Table IVGo).


View this table:
[in this window]
[in a new window]
  		Table IV. Genetic markers exhibiting evidence of linkage to the production of autoantibodies

 
Mapmaker/QTL interval analysis confirmed that regions of chromosomes 1 and 17 showed evidence of linkage by exceeding the threshold set by Lander and Kruglyak for significance (LOD >= 3.3; Fig. 3Go) and also identified regions suggestive of linkage on chromosomes 10 and 16. The pattern of LOD scores produced by the genotype frequencies of loci on chromosome 1 was very complex, with at least three peaks; one between D1Mit12 and D1Mit445 (LOD = 4.3), one proximal to D1Mit396 (LOD = 4.5), and the third between D1Mit396 and D1Mit398 (LOD = 4.5). Almost the whole of chromosome 17 exceeded the threshold for significance, with a maximum LOD occurring at D17Mit83, D17Mit16, and D17Mit24 (LOD = 9.6). Additional regions surpassing the threshold for suggestive linkage (LOD >= 1.9) were identified on chromosomes 10 and 16. Maximum linkage on chromosome 10 was found between the two most proximal markers (D10Mit213 and D10Mit257; LOD = 2.3) and on chromosome 16 between the two most distal markers (D16Mit5 and D16Mit94 LOD = 2.7). The MapManager/QT analysis of the data set confirmed significant linkage (LRS >= 13.2) on chromosomes 1 and 17 and indicated a trend toward linkage (LRS >= 6.8) on chromosome 16 (data not shown).

In summary, all three analyses identified linkage regions controlling susceptibility to autoantibodies in M. bovis-treated NOD mice on chromosomes 1 and 17. The first, here named Babs1 (BCG-induced autoantibodies, locus 1) mapped to proximal chromosome 17 and may be the H2. Linkage on chromosome 1 was complicated as MapManager/QT placed the maximum peak between D1Mit445 and D1Mit396, while the Mapmaker/QTL analysis flanked this region with two other peaks, although the trough between them was not sufficient (i.e., >=1 LOD) to define them as separate loci. Thus, only a single locus could be attributed to chromosome 1, here named Babs2 (BCG-induced autoantibodies, locus 2).

Genotypic and linkage analysis of glomerular immune complex deposition

An autosomal genome scan of 44 female BC1 progeny with glomerular immune complex deposits (by immunofluorescence of C3c) and 52 unaffected female BC1 progeny was conducted using 127 polymorphic microsatellite markers with an average marker separation of 15 cM. Linkage analysis was initially performed at each marker using a 2 x 2 contingency table. No locus exceeded the 95% (significant) experimentwise threshold derived from a permutation analysis of 10,000 iterations of the data ({chi}2 >= 12.0). Genotype frequencies showed a trend suggestive of linkage (37% threshold; {chi}2 >= 6.6) at loci on chromosomes 1, 4, 12, 16, and 17. The NOD alleles contributed susceptibility in a recessive fashion on chromosomes 1, 4, 12, and 17, while the BALB/c allele contributed susceptibility in a dominant fashion on chromosome 16 (Table VGo).


View this table:
[in this window]
[in a new window]
  		Table V. Genetic markers exhibiting evidence of linkage to glomerular immune complex deposition

 
The Mapmaker/QTL and MapManager/QT analyses also failed to identify any genomic regions that exceeded the appropriate thresholds for significant linkage, although regions suggestive of linkage were found on chromosomes 1 and 4. In the Mapmaker/QTL analysis, the region on chromosome 1 exhibiting a trend to linkage was located between D1Mit124 and D1Mit215 at a LOD of 2.4, while that on chromosome 4 occurred between D4Mit170 and D4Mit313 (LOD = 1.9).

In summary, no linkage region controlling susceptibility to glomerular C3c deposition in NOD mice treated with M. bovis surpassed a significant level of linkage with any of the three analyses applied. It is speculated that the existence of background glomerular immune complex deposition in the BALB/c outcross partner reduced the power of this section of the study.


   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Susceptibility to human SLE depends on a multiplicity of genes encoded within the MHC and elsewhere in the genome (reviewed in Ref. 25). In the case of HLA-linked genes, susceptibility to autoimmunity is associated with a limited number of extended haplotypes (supratypes or ancestral haplotypes) that include the class II and class III (complement) genes (26, 27).

The class III genes have been implicated on a number of grounds including: 1) the role of the early complement components in the removal of immune complexes from the circulation, 2) the presence of deletions or silent alleles of C4A and C4B in the susceptible haplotypes such as B8-SC01-DR3-DQ2.1, and 3) the strong association of the rare cases of absolute component deficiencies of C1q, C1r-C1s, or C4 with susceptibility to SLE even though the risks associated with partial C4 deficiency or C2 deficiency are much lower (28). However, linkage disequilibrium with class III null alleles does not fully explain the association of SLE with class II genes within the HLA gene complex (reviewed in Ref. 25), as exemplified by the very strong association between DR4 and hydralazine-induced SLE in women.

In addition to the HLA-linked SLE susceptibility genes, other genetic factors are involved. With the exception of the C1 complement component genes (on chromosome 12p13), which are involved in immune complex handling and are associated with anti-dsDNA autoantibody production and SLE nephritis (29), little is known about them except that they demonstrate a dominant pattern with incomplete penetrance in families (reviewed in Ref. 28) and may include the genes encoding the Ag receptors on T ({alpha}- and {delta}-chains, 14q11.2; ß-chain, 9p22-9cen; {gamma}-chain 7p15-7p14; Ref. 30) and B cells (heavy chain, 14q32; {kappa} light chain, 2p12; {lambda} light chain, 22q11.2; Ref. 31). Three recent genome-wide screens of SLE susceptibility genes in affected pedigrees and affected sib pair families (32, 33, 34) identified two regions significantly (LOD >= 3.6) (20) linked to SLE: 6p11-p21, mapping adjacent to the MHC; and 16q13. These studies also identified six additional regions with LOD scores suggestive of linkage (LOD >= 2.2) (20): 1q41, 1q23, 13q32, 14q21-q23, 20q13, and 20p12; and nine other regions with LOD scores exceeding 1.0.

As with SLE, susceptibility to type 1 diabetes in humans appears to depend on multiple genes encoded within the MHC and elsewhere in the genome (reviewed in Ref. 35). IDDM1 maps to the MHC on chromosome 6p21 and susceptibility is contributed by DRB1, DPB1, DQB1, and DQA1 as well as several other minor genes within the MHC. The contributions of these genes is best seen in the light of ancestral haplotypes; while the incidence of type 1 diabetes in Caucasian communities is around 0.03%, about 6% of individuals with the high-risk MHC haplotype (DR3/DR4) develop diabetes. Linkage to the insulin gene (IDDM2) on 11p15 was also originally identified on the basis of candidature and has been associated with (36), and attributed to (37, 38), allelism in a VNTR locus 5' to the insulin gene, which affects the level of insulin gene expression in the thymus. In addition to IDDM1 and IDDM2, a score of other linkage regions have been identified in genome-wide screens of affected sib pair families (39, 40), although only IDDM4 (11q13), IDDM5 (6q25), and IDDM8 (6q26) have been confirmed in other studies (reviewed in Ref. 35 ; reasons for failure to replicate linkages discussed in Refs. 35 and 41).

The coexistence of SLE and type 1 (autoimmune) diabetes in patients is very rare, and its frequency has never been demonstrated to deviate significantly from that expected to occur by chance alone if the two diseases were completely independent. Similarly, the combination of HA with type 1 diabetes appears to be exceptional. ANA, in contrast, are found in about 15% of children at onset of type 1 diabetes, while are present in only 1% of normal controls (42, 43). Despite the rarity of the coexistence of SLE and type 1 (autoimmune) diabetes, there is good evidence that, at least for loci within the MHC, some alleles confer susceptibility to both diseases. For example, two DR3-associated ancestral haplotypes that confer susceptibility to SLE (B8,SC01,DR3,DQ2,Dw24 and B18,F1C30,DR3,DQ2,Dw25) also confer susceptibility to type 1 diabetes (44). In a recently published controlled correlation of 23 published genome-wide scans of autoimmune or immune-mediated diseases, Becker et al. (8, 9) found that ~65% of positive human linkages mapped nonrandomly into 18 distinct clusters. In addition to the MHC, type 1 diabetes and SLE susceptibility genes colocalized at 7 other IDDM loci, including IDDM2 and IDDM8.

At least three major possibilities could account for these results. First, the 18 clusters identified by Becker could represent areas of the genome in which genes that affect immune function have aggregated through gene duplication and/or selective advantage in a manner similar to the evolution of the MHC. Second, they could represent genes critical to immune function at which different alleles contribute to susceptibility to different immune-related diseases. The third possibility is that certain alleles at each of these loci contribute to susceptibility to autoimmunity per se and that environmental, stochastic, and other genetic factors affect the final phenotypic expression of that tendency to autoimmunity. There are at least two possible ways to directly test the last possibility (the "common gene" hypothesis). The first method involves identifying the alleles responsible for susceptibility to each disease for at least one locus. To date, this has not been achieved, although the possibility that IDDM11 on 14q (45) and the SLE linkage in the same region (33) represents association with the TCR {alpha}-chain gene provides a potential opportunity. The second method is to identify the environmental or genetic modifiers acting on a discrete locus. The model presented here offers an opportunity to identify in NOD mice, candidate regions conferring susceptibility to both diabetes and SLE, depending on exposure to the mycobacterial trigger, with a view to their isolation by congenesis.

Evidence of colocalization of autoimmune disease susceptibility genes in animal models was first reported by Vyse and Todd (10). In a review, they correlated the linkage data from a score of linkage studies of autoimmune diseases in mice and reported that three of nine murine SLE genes and two of three orchitis genes colocalized with diabetes genes, events unlikely to have occurred by chance. Further correlative support has come from independent linkage analyses published since 1996 that have continued to identify further genomic regions associated with susceptibility to multiple autoimmune diseases. For example, Silveira et al. (46) mapped gastritis in mice following neonatal thymectomy to two loci on distal mouse chromosome 4: Gasa1 and Gasa2. Gasa1 mapped to the same genomic region as the SLE susceptibility gene Nba1 (47) and the diabetes susceptibility gene Idd11 (13), and Gasa2 mapped to the same region as the diabetes susceptibility gene Idd9 (48). Support for the common gene hypothesis was also provided by two hypothetico-inductive studies. In the first, Teuscher et al. (49) sought a relationship between the susceptibility of BALB/cBy to two immunization-induced autoimmune diseases: experimental allergic encephalomyelitis (EAE) and experimental allergic orchitis. They studied cosegregation of susceptibility alleles within a BC1 generation by assaying the male progeny of a subsequent backcross for experimental allergic orchitis and the female progeny for EAE. These results demonstrated that the loci conferring susceptibility were probably linked and were consistent with the same locus conferring susceptibility to both diseases. Perhaps the most compelling study was that by Encinas et al. (50), in which they examined diabetes-resistant congenic lines of NOD for susceptibility to EAE. One NOD mouse line, NOD.B6-Idd3, which carried a 0.15-cM region of B6 genome encoding the Il2 gene, was resistant to EAE induction while wild-type NOD mice were susceptible to disease. This result suggested that NOD sequences in this region, perhaps those of the NOD Il2 allele itself, confer susceptibility to the spontaneous autoimmune disease type 1 diabetes as well as to the experimentally induced autoimmune disease EAE.

Here, we have mapped genetic susceptibility to SLE induced in NOD mice by injection with M. bovis, and it is probably of value to consider these data in the light of the common gene hypothesis. Diabetes in NOD mice and SLE in BXSB, NZB/W, and related strains have been extensively mapped, and the regions of apparent colocalization of susceptibility genes reported (10, 11). Restriction of these candidate regions to those which either surpassed Lander and Kryglyak’s thresholds for significance, had been replicated in other studies, or confirmed by congenesis, identified three major genomic regions contributing susceptibility to both diseases: Idd1/Lbw1/Sle4 on chromosome 17, Idd11/Sle2/Nba1/Lbw2 on chromosome 4, and Idd7/Lbw5/Sle3 on chromosome 7. Both lupus and diabetes susceptibility have also been mapped to mouse chromosome 1, although the 95% confidence interval of the chromosome 1 diabetes susceptibility gene Idd5 (51) mapped at least 20 cM proximal to the SLE susceptibility genes identified with the NZB/NZW strains (Sle1/Nba2/Lbw7) (52). In contrast, of the three SLE susceptibility genes mapped to chromosome 1 with the BXSB model, one of these (Bxs1) maps to a similar region as Idd5 while another (Bxs3) maps close to the Sle1/Nba2/Lbw7 cluster of SLE susceptibility genes (53). Therefore, it is possible that the BXSB strain contains both sets of autoimmunity genes on chromosome 1.

We considered the hypothesis that the NOD alleles of genes in the implicated regions of chromosomes 17, 4, and 7 conferred susceptibility to both diabetes and SLE and would therefore be identified in a linkage study of SLE in NOD mice. The first of these regions, that on chromosome 17, was found to be strongly linked to HA (Bah1), ANA (Bana1), and autoantibodies (Babs1) in this study. The strongest association was with autoantibodies for which a {chi}2 of 42.2 (p < 8.08E-11) was obtained at the MHC. This result is consistent with the critical role of the MHC in SLE in humans and the NZB/W (22, 54, 55) and BXSB (56) models and in diabetes in humans, NOD mice (57) and BB rats (58) (Fig. 4Go). Here, homozygosity for the NOD allele of the MHC conferred maximum susceptibility to lupus, whereas in the NZB/W-related strains heterozygosity conferred greatest risk (22, 55, 59, 74, 80) and in the BXSB strain either homozygosity or heterozygosity conferred equal risk (56). Analogous to the situation with DR3 in human autoimmunity, the same H2 haplotype that is associated with susceptibility to diabetes conferred susceptibility to SLE in NOD mice. Therefore, this finding supports the common gene hypothesis, at least as applied to the MHC. In contrast, significant linkage to SLE in this model was not found on proximal chromosome 1, chromosome 4, or chromosome 7 (or indeed any other non-MHC diabetes susceptibility locus). One possible explanation for this finding is that BALB/c mice may carry susceptibility alleles at these loci, and as a result may not be segregating in this cross. There is some experimental support for the idea that BALB/c mice carry autoimmunity susceptibility alleles on distal chromosome 4 as we have mapped to BALB/c gastritis susceptibility alleles to this region (46). Although this problem could have been avoided by using C57BL/6 mice as breeding partners in these crosses, this choice was precluded by the finding reported here of extensive glomerular immune complex deposition in F1 crosses between NOD and B6 mice.



View larger version (18K):
[in this window]
[in a new window]
  		FIGURE 4. Locations of genomic regions conferring susceptibility to murine type 1 diabetes and SLE. Most susceptibility loci are shown as 20-cM regions. Type 1 diabetes loci are designated Idd and are represented by bars containing diagonal lines (13 48 50 51 57 62 63 64 65 66 67 68 69 70 71 72 73 ). All SLE loci are represented by black bars. Loci from NZB/NZW crosses are designated Nba, Sle, and Lbw (47 54 55 59 60 74 ). HA loci from NZB crosses are designated Aem (75 ). Loci from BXSB crosses are designated Bxs (53 56 80 ). Loci from MRL-lpr crosses are designated Lpr, Lmb, and Ldrm (76 77 78 79 ). Loci derived in the study of BCG-induced lupus are represented by white bars and are designated Bah, Bana, and Babs.

 
This study also identified two other regions that have been previously reported to be involved in other autoimmune diseases. The first, a region controlling susceptibility to ANA and autoantibodies on chromosome 1 (Bana3 and Babs2 respectively), lies outside the 95% confidence interval of Idd5, which maps 40 cM proximal of Bana3 and 20 cM proximal of Babs2. However, it maps to the same area as genes controlling susceptibility to ANA in three models of SLE related to the NZB/W model (Fig. 4Go). Thus, linkage to ANA on distal chromosome 1 was found in an NZM/Aeg backcross (Sle1; Ref. 59), a segregating intercross of NZB F1 mice with NZW mice (Nba2; Ref. 47), and an F2 intercross of NZB and NZW mice (Lbw7; Ref. 54). It is of particular interest that this region contained the only NZB/W lupus susceptibility locus to be identified in crosses with both the C57BL/6J and BALBcJ resistant strains (60). Therefore, the study reported here further confirms the major importance of this region in conferring susceptibility to ANA, despite significant differences between the nature of the models and mouse strains involved. The relevance to human disease of this robust linkage is strongly reinforced by genome-wide screens that have identified linkage of lupus to the syntenic region of human chromosome 1, 1q23, (32, 33).

The second region identified in this study was that on chromosome 16 (Bah2) at which BALB/c sequences conferred susceptibility to HA. The genetic marker with strongest linkage to HA on chromosome 16 used in this study (D16Mit58) has previously been identified as showing the strongest linkage to neonatal thymectomy-induced autoimmune ovarian dysgenesis (Aod1; Ref. 61).

The only other genomic region controlling susceptibility to autoimmune disease identified in this study was that on proximal chromosome 10 (Bana2) at which the BALB/c allele also conferred susceptibility to disease. Significant linkage of this locus to any autoimmune disease has not been previously identified, in contrast to the three other genomic regions associated with SLE in this study.

In summary, with the exception of the H2, this study failed to provide direct support for the common gene hypothesis, as the non-MHC-linked loci identified as conferring susceptibility to SLE did not colocalize with those previously implicated in diabetes. The NOD haplotype at the H2 conferred susceptibility to both type 1 (autoimmune) diabetes and SLE, and therefore this region remains a locus for which a congenic approach may allow further study of the interaction between mycobacterial exposure and the expression of autoimmune disease. A striking finding was the nonrandom localization of SLE susceptibility genes, which in three of four cases mapped to regions previously implicated in autoimmunity, thus confirming previous observations of a similar nature. The significance of this clustering remains to be fully explained, but may reflect the genomic structure of loci impacting on immunological function.


   Acknowledgments
 
We thank Karen Knight and Carolyn Pemberton for animal care and Anne-Marie Vachot for assistance.


   Footnotes
 
1 This work was supported by the National Health and Medical Research Council of Australia (NHMRC) and an equipment grant from The Rebecca L. Cooper Medical Foundation. A.G.B. is a recipient of an R. Douglas Wright Fellowship from the NHMRC. P.A.S. is a recipient of an Australian Postgraduate Research Award. Back

2 M.A.J. and P.A.S. contributed equally to this study. Back

3 Address correspondence and reprint requests to Dr. Alan Baxter, Centenary Institute of Cancer Medicine and Cell Biology, Locked bag 6, Newtown NSW 2042, Australia. Back

4 Abbreviations used in this paper: SLE, systemic lupus erythematosus; NOD, nonobese diabetic; HA, hemolytic anemia; ANA, antinuclear autoantibodies; ARD, autoimmune rheumatic disease; BCG, bacillus Calmette-Guérin; LRS, likelihood ratio statistic; LOD, logarithm of odds; QTL, quantitative trait locus; EAE, experimental allergic encephalomyelitis; Sm, Smith; cM, centiMorgan. Back

Received for publication December 27, 1999. Accepted for publication May 19, 2000.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Gourley, I. S., C. C. Patterson, A. L. Bell. 1997. The prevalence of systemic lupus erythematosus in Northern Ireland. Lupus 6:399.[Abstract/Free Full Text]
  2. Johnson, A. E., C. Gordon, F. D. Hobbs, P. A. Bacon. 1996. Undiagnosed systemic lupus erythematosus in the community. Lancet 347:367.[Medline]
  3. Lahita, R. G.. 1995. Special report: adjusted lupus prevalence. Results of a marketing study by the Lupus Foundation of America. Lupus 4:450.[Abstract/Free Full Text]
  4. Hotchberg, M. C.. 1987. The application of genetic epidemiology to systemic lupus erythematosus. J. Rheumatol. 14:867.[Medline]
  5. Baxter, A. G., A. Cooke. 1994. Peptide therapy in diabetes. Lancet 343:1169.
  6. Baxter, A. G., A. C. Horsfall, D. Healey, P. Ozegbe, S. Day, D. G. Williams, A. Cooke. 1994. Mycobacteria precipitate SLE-like syndrome in diabetes prone NOD mice. Immunology 83:227.[Medline]
  7. Horsfall, A. C., R. Howson, P. A. Silveira, D. G. Williams, A. G. Baxter. 1998. Characterization and specificity of B cell responses in lupus induced by M. bovis in NOD/Lt mice. Immunology 95:8.[Medline]
  8. Becker, K. G., R. M. Simon, J. E. Bailey-Wilson, B. Freidlin, W. E. Biddison, H. F. McFarland, J. M. Trant. 1998. Clustering of non-major histocompatibility complex susceptibility candidate loci in human autoimmune diseases. Proc. Natl. Acad. Sci. USA 95:9979.[Abstract/Free Full Text]
  9. Becker, K. G.. 1999. Comparative genetics of type 1 diabetes and autoimmune disease: common loci, common pathways?. Diabetes 48:1353.[Abstract]
  10. Vyse, T. J., J. A. Todd. 1996. Genetic analysis of autoimmune disease. Cell 85:311.[Medline]
  11. Baxter, A. G.. 1997. Immunogenetics and the cause of autoimmune disease. Autoimmunity 25:177.[Medline]
  12. Dietrich, W. F., J. C. Miller, R. G. Steen, M. Merchant, D. Damron, R. Nahf, A. Gross, D. C. Joyce, M. Wessel, R. D. Dredge, et al 1994. A genetic map of the mouse with 4,006 simple sequence length polymorphisms. Nat. Genet. 7:220.[Medline]
  13. Morahan, G., P. McClive, D. Huang, P. Little, A. G. Baxter. 1994. Genetic and physiological association of diabetes susceptibility with raised Na+/H+ exchange activity. Proc. Natl. Acad. Sci. USA 91:5898.[Abstract/Free Full Text]
  14. Lander, E. S., P. Green, J. Abrahamson, A. Barlow, M. J. Daly, S. E. Lincoln, L. Newburg. 1987. MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174.[Medline]
  15. Dietrich, W. F., J. Miller, R. Steen, M. A. Merchant, D. Damron-Boles, Z. Husain, R. Dredge, M. J. Daly, K. A. Ingalls, J. T. O’Connor, et al 1996. A comprehensive genetic map of the mouse genome. Nature 380:149.[Medline]
  16. Churchill, G. A., R. W. Doerge. 1994. Empirical threshold values for quantitative trait mapping. Genetics 138:963.[Abstract]
  17. Manly, K. F.. 1993. A Macintosh program for storage and analysis of experimental genetic mapping data. Mamm. Genome 4:303.[Medline]
  18. Visscher, P. M., C. S. Haley, S. A. Knott. 1996. Mapping QTLs for binary traits in backcross and F2 populations. Gen. Res. 68:55.
  19. Haley, C. S., S. A. Knott. 1992. A simple regression method for mapping quantitative trait loci in line crosses using flanking markers. Heredity 69:315.[Medline]
  20. Lander, E., L. Kruglyak. 1995. Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results. Nat. Genet. 11:241.[Medline]
  21. Lander, E. S., D. Botstein. 1989. Mapping mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121:185.[Abstract/Free Full Text]
  22. Morel, L., U. H. Rudofsky, J. A. Longmate, J. Schiffenbauer, E. K. Wakeland. 1994. Polygenic control of susceptibility to murine systemic lupus erythematosus. Immunity 1:219.[Medline]
  23. Eilat, D., Y. Naparstek. 1999. Anti-DNA autoantibodies: a puzzle of autoimmune phenomena. Immunol. Today 20:339.[Medline]
  24. Lefkowith, J. B., G. S. Gilkeson. 1996. Nephritogenic autoantibodies in lupus. Arthritis Rheum. 39:894.[Medline]
  25. Vyse, T. J., B. L. Kotzin. 1998. Genetic susceptibility to systemic lupus erythematosus. Annu. Rev. Immunol. 16:261.[Medline]
  26. Kelly, H., V. J. McCann, P. H. Kay, R. L. Dawkins. 1985. Susceptibility to IDDM is marked by MHC supratypes rather than individual alleles. Immunogenetics 22:643.[Medline]
  27. Hartung, K., M. P. Baur, R. Coldewey, M. Fricke, J. R. Kalden, H. J. Lakomek, H. H. Peter, D. Schendel, P. M. Schneider, S. A. Seuchter, et al 1992. Major histocompatibility complex haplotypes and complement C4 alleles in systemic lupus erythematosus: results of a multicenter study. J. Clin. Invest. 90:1346.
  28. Arnett, F. C., J. D. Reveille. 1992. Genetics of systemic lupus erythematosus. Rheum. Dis. Clin. N. Am. 18:865.[Medline]
  29. Walport, M. J., P. J. Lachman. 1990. Complement deficiencies and abnormalities of the complement system in systemic lupus erythematosus and related disorders. Curr. Opin. Rheumatol. 2:661.[Medline]
  30. Tebib, J. G., J. Alcocer-Varela, D. Alarcon-Segovia, P. H. Schur. 1990. Association between a T cell receptor restriction length polymorphism and systemic lupus erythematosus. J. Clin. Invest. 86:1961.
  31. Olee, T., P. M. Yang, K. A. Siminovitch, N. J. Olsen, J. Hillson, J. Wu, F. Kozin, D. A. Carson, P. P. Chen. 1991. Molecular basis of an autoantibody-associated restriction fragment length polymorphism that confers susceptibility to autoimmune disease. J. Clin. Invest. 88:193.
  32. Moser, K. L., B. R. Neas, J. E. Salmon, H. Yu, N. Gray-McGuire, G. R. Asundi, J. Bruner, J. Fox, S. Henshall Kelly, et al 1998. Genome scan of human systemic lupus erythematosus: evidence for linkage on chromosome 1q in African-American pedigrees. Proc. Natl. Acad. Sci. USA 95:14869.[Abstract/Free Full Text]
  33. Gaffney, P. M., G. M. Kearns, K. B. Shark, W. A. Ortmann, S. A. Selby, M. L. Malmgren, K. E. Rohlf, T. C. Ockenden, R. P. Messner, R. A. King, S. S. Rich, T. W. Behrens. 1998. A genome-wide search for susceptibility genes in human systemic lupus erythematosus sib-pair families. Proc. Natl. Acad. Sci. USA 95:14875.[Abstract/Free Full Text]
  34. Shai, R., Jr F. P. Quismorio, L. Li, O. J. Kwon, J. Morrison, D. J. Wallace, C. M. Neuwelt, C. Brautbar, W. j. Gauderman, C. O. Jacob. 1999. Genome-wide screen for systemic lupus erythematosus susceptibility genes in multiplex families. Hum. Mol. Genet. 8:639.[Abstract/Free Full Text]
  35. She, J.-X., M. P. Marron. 1998. Genetic susceptibility factors in type 1 diabetes: linkage, disequilibrium and functional analyses. Curr. Opin. Immunol. 10:682.[Medline]
  36. Julier, C., R. N. Hyer, J. Davies, F. Merlin, P. Soularue, L. Briant, G. Cathelineau, I. Deschamps, J. I. Rotter, P. Froguel, C. Boitard, J. I. Bell, G. M. Lathrop. 1991. Insulin-IGF2 region on chromosome 11p encodes a gene implicated in HLA-DR4-dependent diabetes susceptibility. Nature 354:155.[Medline]
  37. Vafiadis, P., S. Y. Bennett, J. A. Todd, J. Nadeau, R. Grabs, C. G. Goodyer, S. Wickramasinghe, E. Colle, C. Polychronakos. 1997. Insulin expression in human thymus is modulated by INS VNTR alleles at the IDDM2 locus. Nat. Genet. 15:289.[Medline]
  38. Pugliese, A., M. Zeller, A. Fernandez, L. J. Zalcberg, R. J. Bartlett, C. Ricordi, M. Pietropaolo, G. S. Eisenbarth, S. T. Bennet, D. D. Patel. 1997. The insulin gene is transcribed in the human thymus and transcription levels correlate with allelic variation at the INS VNTR-IDDM2 susceptibility locus for type 1 diabetes. Nat. Genet. 15:293.[Medline]
  39. Mein, C. A., L. Esposito, M. G. Dunn, G. C. L. Johnson, A. E. Timms, J. V. Goy, A. N. Smith, L. Sebag-Montefiore, M. E. Merriman, A. J. Wilson, et al 1998. A search for type 1 diabetes susceptibility genes in families from the United Kingdom. Nat. Genet. 19:297.[Medline]
  40. Concannon, P., K. J. Gogolin-Ewens, D. A. Hinds, B. Wapelhorst, V. A. Morrison, B. Stirling, M. Mitra, J. Farmer, S. R. Williams, N. J. Cox, et al 1998. A second generation screen of the human genome for susceptibility to insulin-dependent diabetes mellitus. Nat. Genet. 19:292.[Medline]
  41. Lernmark, A., J. Ott. 1998. Sometimes it’s hot, sometimes it’s not. Nat. Genet. 19:213.[Medline]
  42. Notsu, K., S. Note, N. Nabeya, S. Kuno, T. Sakurami. 1983. Antinuclear antibodies in childhood diabetics. Endocrinologia Japonica 30:469.[Medline]
  43. Menser, M. A., J. R. Hudson. 1983. Pancreatic islet and other autoantibodies in juvenile and adult onset diabetics in Australia. Pathology 15:309.[Medline]
  44. Segurado, O. G., P. Iglesias-Casarrubios, J. Martinez-Laso, A. Corell, J. M. Martin-Villa, A. Arnaiz-Villena. 1991. Autoimmunogenic HLA-DRB1*0301 allele (DR3) may be distinguished at the DRB1 non-coding regions of HLA-B8,DR3,Dw24 and B18,DR3,Dw25 haplotypes. Mol. Immunol. 28:189.[Medline]
  45. Field, L. L., R. Tobias, G. Thomson, S. Plon. 1996. Susceptibility to insulin-dependent diabetes mellitus maps to a locus (IDDM11) on human chromosome 14q24.3-q31. Genomics 33:1.[Medline]
  46. Silveira, P. A., A. G. Baxter, W. E. Cain, I. R. van Driel. 1999. A major linkage region on distal chromosome 4 confers susceptibility to mouse autoimmune gastritis. J. Immunol. 162:5106.[Abstract/Free Full Text]
  47. Drake, C. G., S. K. Babcock, E. Palmer, B. L. Kotzin. 1994. Genetic analysis of the NZB contribution to lupus-like autoimmune disease in (NZB x NZW)F1 mice. Proc. Natl. Acad. Sci. USA 91:4062.[Abstract/Free Full Text]
  48. Rodrigues, N. R., R. J. Cornall, P. Chandler, E. Simpson, L. S. Wicker, L. B. Peterson, J. A. Todd. 1994. Mapping of an insulin-dependent diabetes locus, Idd9, in NOD mice to chromosome 4. Mamm. Genome 5:167.[Medline]
  49. Teuscher, C., W. F. Hickey, C. M. Grafer, K. S. Tung. 1998. A common immunoregulatory locus controls susceptibility to actively induced experimental allergic encephalomyelitis and experimental allergic orchitis in BALB/c mice. J. Immunol. 160:2751.[Abstract/Free Full Text]
  50. Encinas, J. A., L. S. Wicker, L. B. Peterson, A. Mukasa, C. Teuscher, R. Sobel, H. L. Weiner, C. E. Seidman, J. G. Seidman, V. K. Kuchroo. 1999. QTL influencing autoimmune diabetes and encephalomyelitis map to a 0.15-cM region containing IL2. Nat. Genet. 21:158.[Medline]
  51. Cornall, R. J., J. B. Prins, J. A. Todd, A. Pressey, N. H. DeLarato, L. S. Wicker, L. B. Peterson. 1991. Type 1 diabetes in mice is linked to the interleukin-1 receptor and Lsh/Ity/Bcg genes on chromosome 1. Nature 353:262.[Medline]
  52. Blake, J. A., J. E. Richardson, M. T. Davisson, J. T. Eppig. 1999. The Mouse Genome Database (MGD): genetic and genomic information about the laboratory mouse. The Mouse Genome Database Group. Nucleic Acids Res. 27:95.[Abstract/Free Full Text]
  53. Hogarth, M. B., J. H. Slingsby, P. J. Allen, E. M. Thompson, P. Chandler, K. A. Davies, E. Simpson, B. J. Morley, M. J. Walport. 1998. Multiple lupus susceptibility loci map to chromosome 1 in BXSB mice. J. Immunol. 161:2753.[Abstract/Free Full Text]
  54. Kono, D. H., R. W. Burlingame, D. G. Owens, A. Kuramochi, R. S. Balderas, D. Balomenos, A. N. Theofilopoulos. 1994. Lupus susceptibility loci in New Zealand mice. Proc. Natl. Acad. Sci. USA 91:10168.[Abstract/Free Full Text]
  55. Vyse, T. J., L. Morel, F. J. Tanner, E. K. Wakeland, B. L. Kotzin. 1996. Backcross analysis of genes linked to autoantibody production in New Zealand White mice. J. Immunol. 157:2719.[Abstract]
  56. Merino, R., L. Fossati, M. Lacour, R. Lemoine, M. Higaki, S. Izui. 1992. H-2-linked control of the Yaa gene-induced acceleration of lupus-like autoimmune disease in BXSB mice. Eur. J. Immunol. 22:295.[Medline]
  57. Hattori, M., J. B. Buss, R. A. Jackson, L. Glimcher, M. E. Dorf, M. Minami, S. Makino, K. Moriwaki, H. Kuzuya, H. Imura, et al 1986. The NOD mouse: recessive diabetogenic gene in the major histocompatibility complex. Science 231:733.[Abstract/Free Full Text]
  58. Colle, E., R. D. Guttmann, T. A. Seemayer. 1981. Spontaneous diabetes mellitus syndrome in the rat. I. Association with the major histocompatibility complex. J. Exp. Med. 154:1237.[Abstract/Free Full Text]
  59. Morel, L., C. Mohan, Y. Yu, B. P. Croker, N. Tian, A. Deng, E. K. Wakeland. 1997. Functional dissection of systemic lupus erythematosus using congenic mouse strains. J. Immunol. 158:6019.[Abstract]
  60. Rozzo, S. J., T. J. Vyse, C. G. Drake, B. L. Kotzin. 1996. Effect of genetic background on the contribution of New Zealand Bloack loci to autoimmune lupus nephritis. Proc. Natl. Acad. Sci. USA 93:15164.[Abstract/Free Full Text]
  61. Wardel, B. B., S. D. Michael, K. S. K. Tung, J. A. Todd, E. P. Blankenhorn, K. McEntee, J. D. Sudweeks, W. K. Hansen, N. D. Meker, J. S. Griffith, K. D. Livingstone, C. Teuscher. 1995. Aod1 the immunoregulatory locus controlling abrogation of tolerance in neonatal thymectomy-induced autoimmune ovarian dysgenesis, maps to mouse chromosome 16. Proc. Natl. Acad. Sci. USA 924758..
  62. Prochazka, M., E. H. Leiter, D. V. Serreze, D. L. Coleman. 1987. Three recessive loci required for insulin-dependent diabetes in nonobese diabetic mice. Science 237:286.[Abstract/Free Full Text]
  63. Wicker, L. S., M. C. Appel, F. Dotta, A. Pressey, B. J. Miller, N. H. DeLarato, P. A. Fischer, Jr R. C. Boltz, L. B. Peterson. 1992. Autoimmune syndromes in major histocompatibility complex (MHC) congenic strains of nonobese diabetic (NOD) mice: the NOD MHC is dominant for insulitis and cyclophosphamide-induced diabetes. J. Exp. Med. 176:67.[Abstract/Free Full Text]
  64. McAleer, M. A., P. Reifsnyder, S. M. Palmer, M. Prochazka, J. M. Love, J. B. Copeman, E. E. Powell, N. R. Rodrigues, J. B. Prins, D. V. Serreze. 1995. Crosses of NOD mice with the related NON strain: a polygenic model for IDDM. Diabetes 44:1186.[Abstract]
  65. Todd, J. A., T. J. Aitman, R. J. Cornall, S. Ghosh, J. R. Hall, C. M. Hearne, A. M. Knight, J. M. Love, M. A. McAleer, J. B. Prins. 1991. Genetic analysis of autoimmune type 1 diabetes mellitus in mice. Nature 351:542.[Medline]
  66. Dallas-Pedretti, A., M. McDuffie, K. Haskins. 1995. A diabetes-associated T-cell autoantigen maps to a telomeric locus on mouse chromosome 6. Proc. Natl. Acad. Sci. USA 92:1386.[Abstract/Free Full Text]
  67. Wicker, L. S., J. A. Todd, J. B. Prins, P. L. Podolin, R. J. Renjilian, L. B. Peterson. 1994. Resistance alleles at two non-major histocompatibility complex-linked insulin-dependent diabetes loci on chromosome 3, Idd3 and Idd10, protect nonobese diabetic mice from diabetes. J. Exp. Med. 180:1705.[Abstract/Free Full Text]
  68. Podolin, P. L., P. Denny, N. Armitage, C. J. Lord, N. J. Hill, E. R. Levy, L. B. Peterson, J. A. Todd, L. S. Wicker, P. A. Lyons. 1998. Localization of two insulin-dependent diabetes (Idd) genes to the Idd10 region on mouse chromosome 3. Mamm. Genome 9:283.[Medline]
  69. Brodnicki, T., P. McClive, S. Couper, D. Huang, G. Morahan. 1997. Genetic mapping of a diabetes susceptibility gene on mouse chromosome 4 using congenic mouse strains. Australasian Society for Immunology 27th Annual Meeting, Perth 57. ASI Inc., Melbourne.
  70. Serreze, D. V., M. Prochazka, P. C. Reifsnyder, M. M. Bridgett, E. H. Leiter. 1994. Use of recombinant congenic and congenic strains of NOD mice to identify a new insulin-dependent diabetes resistance gene. J. Exp. Med. 180:1553.[Abstract/Free Full Text]
  71. Ikegami, H., S. Makino, E. Yamato, Y. Kawaguchi, H. Ueda, T. Sakamoto, K. Takekawa, T. Ogihara. 1995. Identification of a new susceptibility locus for insulin-dependent diabetes mellitus by ancestral haplotype congenic mapping. J. Clin. Invest. 96:1936.
  72. Hattori, M., E. Yamato, N. Itoh, H. Senpuku, T. Fujisawa, M. Yoshino, M. Fukuda, E. Matsumoto, T. Toyonaga, I. Nakagawa, et al 1999. Cutting edge: homologous recombination of the MHC class I K region defines new MHC-linked diabetogenic susceptibility gene(s) in nonobese diabetic mice. J. Immunol. 163:1721.[Abstract/Free Full Text]
  73. Denny, P., C. J. Lord, N. J. Hill, J. V. Goy, E. R. Levy, P. L. Podolin, L. B. Peterson, L. S. Wicker, J. A. Todd, P. A. Lyons. 1997. Mapping of the IDDM locus Idd3 to a 0.35-cM interval containing the interleukin-2 gene. Diabetes 46:695.[Abstract]
  74. Vyse, T. J., C. G. Drake, S. J. Rozzo, E. Roper, S. Izui, B. L. Kotzin. 1996. Genetic linkage of IgG autoantibody production in relation to lupus nephritis in New Zealand hybrid mice. J. Clin. Invest. 98:1762.[Medline]
  75. Ochiai, K., S. Ozaki, A. Tanino, S. Watanabe, T. Ueno, K. Mitsui, J. Toei, Y. Inada, S. Hirose, T. Shirai, H. Nishimura. 2000. Genetic regulation of anti-erythrocyte autoantibodies and splenomegaly in autoimmune hemolytic anemia-prone New Zealand Black mice. Int. Immunol. 12:1.[Abstract/Free Full Text]
  76. Watanabe-Fukunaga, R., C. I. Brannan, N. G. Copeland, N. A. Jenkins, S. Nagata. 1992. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature 356:314.[Medline]
  77. Watson, M. L., J. K. Rao, G. S. Gilkeson, P. Ruiz, E. M. Eicher, D. S. Pisetsky, A. Matsuzawa, J. M. Rochelle, M. F. Seldin. 1992. Genetic analysis of MRL-lpr mice: relationship of the Fas apoptosis gene to disease manifestations and renal disease-modifying loci. J. Exp. Med. 176:1645.[Abstract/Free Full Text]
  78. Wang, Y., M. Nose, T. Kamoto, M. Nishimura, H. Hiai. 1997. Host modifier genes affect mouse autoimmunity induced by the lpr gene. Am. J. Pathol. 151:1791.[Abstract]
  79. Vidal, S., D. H. Kono, A. N. Theofilopoulos. 1998. Loci predisposing to autoimmunity in MRL-Faslpr and C57BL/6-Faslpr mice. J. Clin. Invest. 101:696.[Medline]
  80. Kawano, H., M. Abe, D. Zhang, T. Saikawa, M. Fujimori, S. Hirose, T. Shirai. 1992. Heterozygosity of the major histocompatibility complex controls the autoimmune disease in (NZW x BXSB)F1 mice. Clin. Immunol. Immunopathol. 65:308.[Medline]



This article has been cited by other articles:


Home page
 J. Immunol.Home page
J. M. Fletcher, M. A. Jordan, S. L. Snelgrove, R. M. Slattery, F. D. Dufour, K. Kyparissoudis, G. S. Besra, D. I. Godfrey, and A. G. Baxter
Congenic Analysis of the NKT Cell Control Gene Nkt2 Implicates the Peroxisomal Protein Pxmp4
J. Immunol., September 1, 2008; 181(5): 3400 - 3412.
[Abstract] [Full Text] [PDF]

Home page
 Int ImmunolHome page
D. K. Y. Ang, T. C. Brodnicki, M. A. Jordan, W. E. Wilson, P. Silveira, B. L. Gliddon, A. G. Baxter, and I. R. van Driel
Two genetic loci independently confer susceptibility to autoimmune gastritis
Int. Immunol., September 1, 2007; 19(9): 1135 - 1144.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
M. A. Jordan, J. M. Fletcher, D. Pellicci, and A. G. Baxter
Slamf1, the NKT Cell Control Gene Nkt1
J. Immunol., February 1, 2007; 178(3): 1618 - 1627.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
J. I. Elliott, A. Sardini, J. C. Cooper, D. R. Alexander, S. Davanture, G. Chimini, and C. F. Higgins
Phosphatidylserine exposure in B lymphocytes: a role for lipid packing
Blood, September 1, 2006; 108(5): 1611 - 1617.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
A.-C. Rocha-Campos, R. Melki, R. Zhu, N. Deruytter, D. Damotte, M. Dy, A. Herbelin, and H.-J. Garchon
Genetic and Functional Analysis of the Nkt1 Locus Using Congenic NOD Mice: Improved V{alpha}14-NKT Cell Performance but Failure to Protect Against Type 1 Diabetes.
Diabetes, April 1, 2006; 55(4): 1163 - 1170.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
M. Lundholm, V. Motta, A. Lofgren-Burstrom, N. Duarte, M.-L. Bergman, S. Mayans, and D. Holmberg
Defective Induction of CTLA-4 in the NOD Mouse Is Controlled by the NOD Allele of Idd3/IL-2 and a Novel Locus (Ctex) Telomeric on Chromosome 1
Diabetes, February 1, 2006; 55(2): 538 - 544.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
S. Koarada, Y. Wu, N. Fertig, D. A. Sass, M. Nalesnik, J. A. Todd, P. A. Lyons, J. Fenyk-Melody, D. B. Rainbow, L. S. Wicker, et al.
Genetic Control of Autoimmunity: Protection from Diabetes, but Spontaneous Autoimmune Biliary Disease in a Nonobese Diabetic Congenic Strain
J. Immunol., August 15, 2004; 173(4): 2315 - 2323.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
H. Fan, A. Longacre, F. Meng, V. Patel, K. Hsiao, J. S. Koh, and J. S. Levine
Cytokine Dysregulation Induced by Apoptotic Cells Is a Shared Characteristic of Macrophages from Nonobese Diabetic and Systemic Lupus Erythematosus-Prone Mice
J. Immunol., April 15, 2004; 172(8): 4834 - 4843.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
L. M. Esteban, T. Tsoutsman, M. A. Jordan, D. Roach, L. D. Poulton, A. Brooks, O. V. Naidenko, S. Sidobre, D. I. Godfrey, and A. G. Baxter
Genetic Control of NKT Cell Numbers Maps to Major Diabetes and Lupus Loci
J. Immunol., September 15, 2003; 171(6): 2873 - 2878.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
O. Boulard, G. Fluteau, L. Eloy, D. Damotte, P. Bedossa, and H.-J. Garchon
Genetic Analysis of Autoimmune Sialadenitis in Nonobese Diabetic Mice: A Major Susceptibility Region on Chromosome 1
J. Immunol., April 15, 2002; 168(8): 4192 - 4201.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
I. A. Eaves, L. S. Wicker, G. Ghandour, P. A. Lyons, L. B. Peterson, J. A. Todd, and R. J. Glynne
Combining Mouse Congenic Strains and Microarray Gene Expression Analyses to Study a Complex Trait: The NOD Model of Type 1 Diabetes
Genome Res., February 1, 2002; 12(2): 232 - 243.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jordan, M. A.
Right arrow Articles by Baxter, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jordan, M. A.
Right arrow Articles by Baxter, A. G.
Right arrowPubmed/NCBI databases
*Gene
*Substance via MeSH
Medline Plus Health Information
*Diabetes Type 1
*Lupus

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS