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Human Molecular Genetics, 2004, Vol. 13, No. 15 1641-1648
DOI: 10.1093/hmg/ddh179
Human Molecular Genetics, Vol. 13, No. 15 © Oxford University Press 2004; all rights reserved

Significant linkage to spondyloarthropathy on 9q31–34

Corinne Miceli-Richard1,2,*, Habib Zouali1, Roula Said-Nahal2, Suzanne Lesage1, Françoise Merlin1, Claudia de Toma1, Hélène Blanche1, Mourad Sahbatou1, Maxime Dougados2, Gilles Thomas1, Maxime Breban3, Jean-Pierre Hugot1,4 and Groupe Français d'Etude Génétique des Spondylarthropathies (GFEGS)

1Laboratoire de Génétique des Maladies Inflammatoires de l'Intestin et Fondation Jean Dausset/CEPH, Paris, France, 2Hôpital Cochin, Assistance Publique-Hôpitaux de Paris, Université René Descartes Paris, France, 3Hôpital Ambroise Paré, Assistance Publique-Hôpitaux de Paris, Université Paris, Ile de France Ouest Paris, France and 4Equipe Avenir Inserm, Hôpital Robert Debré, Assistance Publique-Hôpitaux de Paris, Faculté Xavier Bichat, Paris, France

Received March 19, 2004; Accepted June 2, 2004


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 RESULTS
 DISCUSSION
 MATERIALS AND METHODS
 REFERENCES
 
Spondyloarthropathy (SpA) is a frequent rheumatologic disorder with a prevalence of 0.3% in Caucasian populations from western Europe. It commonly presents as chronic axial and/or peripheral arthritis with potential disabling outcome. SpA is also variably associated with extra-articular manifestations. The pathogenesis of SpA is considered as complex, with a strong genetic component. Human leukocyte antigen B27 has been identified as a predisposing factor for SpA, but family and twin studies suggest that additional genetic risk factors exist outside the major histocompatibility complex (MHC). To map SpA susceptibility loci, 120 multiplex SpA families were included in a genome-wide scan. Linkage analyses performed on the first 65 families allowed us to identify four candidate non-MHC regions on chromosomes 5q, 9q, 13q and 17q, which were further explored in the remaining 55 multiplex families (extension study). Non-parametric multipoint linkage analyses of the whole data set yielded evidence of significant linkage to 9q31–34, in the vicinity of marker D9S1776 (NPL=4.87, LOD=5.15, P=0.00002). This result provides evidence for the presence of a non-MHC susceptibility locus for SpA mapping to 9q31–34.


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
RESULTS
 DISCUSSION
 MATERIALS AND METHODS
 REFERENCES
 
Spondyloarthropathy (SpA) is one of the commonest chronic rheumatic diseases. Its prevalence has been set recently in western Europe as 0.3% (1), providing evidence that SpA is more prevalent than previously thought. This observation also suggests that the potential disabling outcome of this chronic inflammatory condition has a clear socioeconomic impact.

SpA includes a spectrum of related disorders comprising the prototype ankylosing spondylitis (AS), a subset of psoriatic arthritis (PsA), reactive arthritis (ReA), arthritis associated with inflammatory bowel disease (IBD), and undifferentiated SpA (USpA) which are difficult to differentiate because they may occur simultaneously or sequentially in the same patient. A critical question regarding the SpA spectrum is to determine to what extent predisposing genetic factors, other than HLA-B27, are common to the distinct forms of SpA. This question has been addressed previously through a phenotypic study of multiplex SpA families, concluding that distinct subsets of SpA should be considered as various phenotypic expressions of the same disease (2,3), thereby justifying the SpA disease affection status that we have used herein.

The human leukocyte antigen (HLA) B27 has been the first genetic factor identified in AS (4,5). The direct involvement of the HLA-B27 allele in the physiopathology of the disease (rather than a nearby alternative allele in linkage disequilibrium with HLA-B27 for example) is supported by the occurrence of peripheral and axial arthritis, gut inflammation, genital and skin lesions in HLA-B27 transgenic rats (6). However, the exact mechanism by which the B27 molecule may induce a chronic inflammation is still a matter of debate.

Family and twin studies have further demonstrated that the predisposition to the disease was not exclusively related to HLA-B27, suggesting that additional susceptibility genes are expected in SpA. In fact, concordance rates for HLA-B27-positive monozygotic twins (50–63%) differ highly from concordance rates for HLA-B27-positive dizygotic twins (20–27%) (7,8). Moreover, a sibling risk ratio ({lambda}S) value of 82 has been estimated (9,10). In comparison, the {lambda}HLA value ranges from 5 to 6 according to the data from genome-wide ASP linkage analysis (11). Such results provide evidence for the involvement of genetic factors arising from outside the major histocompatibility complex (MHC) (12).

Several groups, including ourselves, have been involved in the search for susceptibility genes by whole-genome screening and/or candidate gene approaches. Using the candidate gene approach, weak associations between SpA and CYP2D6 (13) and ANKH (14) have been reported, but the most convincing associations arise from the study of a positional candidate gene, located in a region where a suggestive linkage was previously reported, on chromosome 2q (11): interleukine-1 receptor antagonist (15,16). Using the alternative positional cloning strategy, the Oxford group has reported two whole genome studies on British Caucasian populations (11,17). In addition to the MHC, these screens have evidenced suggestive linkage on chromosomes 1p, 2q, 9q, 10q, 16q and 19q. The strongest linkage observed outside the MHC was on chromosome 16q, with the maximum linkage observed at 101 cM from the p-telomere (NPL score of 4.7).

In order to replicate these localizations and to identify additional susceptibility loci, we initiated linkage studies on 120 SpA French Caucasian families.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 RESULTS
DISCUSSION
 MATERIALS AND METHODS
 REFERENCES
 
Genome screen
Genome-wide linkage screen was performed for 65 families (first step) containing 180 affected individuals and 268 genotyped relatives. The results of the linkage screen are displayed in Figure 1. Twelve markers yielded NPL values greater than 2.2 (P<0.01) corresponding to five chromosomal regions including the MHC locus on chromosome 6p (Table 1). As expected, the highest NPL score was found on chromosome 6p22 with a significant linkage observed near marker D6S276 (max NPL score at 29 cM from p-telomere NPL=5.29, LOD=6.07, P=0.0000001), a marker related to the MHC locus (z0=0.04, z1=0.45, z2=0.51). The calculated {lambda}S value at this locus was 6.25. The markers yielding the most significant results in the four other regions (5q, 9q, 13q and 17q) were D5S471 (NPL=2.55, LOD=1.41, P=0.01), D9S1682 (NPL=2.32, LOD=1.17, P=0.009), D13S153 (NPL=2.55, LOD=1.41, P=0.005) and D17S949 (NPL=2.44, LOD=1.29, P=0.007). No linkage was observed on chromosome 16q (Fig. 1). However, because this region was previously reported as the best non-MHC region with significant linkage for AS (11,17), we included it in the extension study. Altogether, five regions located on chromosome 5q, 9q, 13q, 16q and 17q, were retained for the extension study.



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  		Figure 1. Multipoint non-parametric linkage analyses (NPL scores) of the whole-genome screening. NPL scores (GENEHUNTER) are shown on the vertical axis; the distance from the p-terminal end of the chromosome (in cM) is shown on the horizontal axis.

 

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  		Table 1. Linkage analysis of the whole-data set (120 families) within the six fully explored regions
 
Extension study and combined analysis
We genotyped 55 additional independent SpA multiplex families for markers centred on the above candidate regions. Linkage analyses performed on all 120 families exhibited neither significant nor suggestive linkage to chromosomes 5q, 13q, 16q or 17q (Table 1). However, significant linkage was observed on chromosome 9, with a maximum NPL score of 4.87 (LOD=5.15, P=0.00002) at 110 cM from the telomere. According to the guidelines for evaluation of results from linkage analyses (18), this value was considered as significant and allowed us to conclude that a SpA susceptibility locus resides on chromosome 9q.

By examining the pedigrees separately, we found that pedigree 110 contributed significantly to the overall linkage score (maximum NPL score for this family at 98 cM from the telomere, P=0.0004). This family included 10 affected relatives until the 5th degree, all sharing a common haplotype on chromosome 9q (Fig. 2). By examining the co-segregation of this haplotype with the disease status within the family, and by taking into account the HLAB27 carrier status, we inferred a possible dominant model of inheritance with incomplete penetrance (Fig. 2).



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  		Figure 2. Pedigree structure of family 110 and transmitted haplotype at the 9q31–34 locus. (A) Family members were genotyped for five microsatellite markers mapping on chromosome 9q. The markers are ordered from centromere to telomere. The inter-marker distances are reported in cM. (B) Affected, unknown and healthy individuals are represented by respective black, grey and white symbols. Rings indicate HLA-B27 carriers. Crosses represent recombination events. Boxes indicate the minimal chromosome regions shared by all affected family members.

 
On the basis of this finding, parametric analyses at the 9q locus were then performed on the 120 families. The model leading to the highest LOD score corresponded to the following penetrances: 0.001 (wild-type homozygotes), 0.100 (heterozygotes) and 0.100 (mutant homozygotes), with a mutant allele frequency of 0.001. Using this model of inheritance, multipoint LOD scores were then maximized for varying fractions of linked families ({alpha}). A maximum HLOD of 3.08 was obtained, with a corresponding {alpha} value of 0.23.


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 RESULTS
 DISCUSSION
MATERIALS AND METHODS
 REFERENCES
 
This genome-wide search for SpA susceptibility genes allowed detection of a significant linkage on chromosome bands 9q31–34.

This study was performed on Caucasian multiplex families with a relatively large number of affected relatives (336 affected relative pairs in 120 families) and distant affected relatives in one-third of families. In the genotyped families, a great proportion of patients had both parents genotyped (72% in sib-pair families), and in case of non-genotyped parents, unaffected relatives allowed to infer the information available for calculation of identity-by-descent in the vast majority of cases. These overall characteristics of our family cohort make it powerful to detect linkage (19).

The diagnosis of affected family members was carefully checked by a qualified rheumatologist specialized in SpA. Many previous genetic studies have been limited to the endophenotype AS (11,17), defined according to the modified New York criteria (20). These criteria imply strict radiographic evidence of advanced sacroiliitis for AS diagnosis. The Amor or the European Spondylarthropathy Study Group (ESSG) criteria used herein (21,22) permit establishment of an SpA diagnosis even at an early stage of the disease, in the absence of radiographic lesions (23). In fact, an incremental frequency of radiographic sacroiliitis over time has been demonstrated in SpA (40% prevalence after 10 years of disease duration, 86% prevalence after 20 years of disease duration), suggesting the inaccuracy of the radiological changes for early diagnosis (2). We can thus consider that the diagnosis criteria chosen herein allowed us to classify as affected a higher proportion of individuals than using the AS phenotype only.

As expected, strong linkage was identified in the MHC region containing the well-known HLA-B risk factor. The calculated {lambda}S value at this locus was 6.25, in accordance with previous results [{lambda}S=5.2 (11)]. No additional locus provided evidence of linkage with such a magnitude, confirming the great importance of the MHC in the disease susceptibility. However, this {lambda}S value was much lower than the overall {lambda}S value of 50, previously estimated in AS (24) and argued in favour of the involvement of other genetic susceptibility factors arising from outside the MHC.

Five additional regions exhibited a weak linkage signal in the first genome-wide step and were further investigated in an extension study. These regions are located on chromosomes 5q, 9q, 13q, 16q and 17q.

The region of interest on chromosome 13q was not previously linked to AS or, to our knowledge, to other closely related conditions. Positive linkage was not confirmed by the extension study suggesting that the weak linkage observed in the genome scan step was a false positive result.

On chromosome 5, the maximum NPL score was found at 122 Mb from the telomere, corresponding to the chromosome band 5q21. This locus is close to a cytokine gene cluster located at 132 Mb from the telomere. Interestingly, this region also contains a locus (IBD5) that confers susceptibility to Crohn's disease (CD), a condition closely related to SpA (25). Unfortunately, the extension study failed to confirm a linkage in this region. The IBD phenotype was segregating only among 12 of the whole family set. Therefore, analyses on this phenotype subgroup were not performed.

On chromosome 17, the extension study also failed to confirm a positive linkage. However, the region identified on chromosome 17q (maximum NPL score at 88 cM from the telomere) was relatively close to the locus, PSOR2, mapped to the interval 112–180 cM from the p-telomere and which confers susceptibility to psoriasis (26–28), another condition associated with SpA. Despite of the negative results, this relative proximity prompted us to further analyze this region in PsA families. When looking at the families demonstrating a positive linkage on chromosome 17q, we did not observe any excess of families where psoriasis was segregating (data not shown). Moreover, complementary linkage analyses were performed on the subset of 21 families where psoriasis was segregating. We observed that the linkage signal fell down dramatically (NPL score=0.81; LOD=0.14, P=0.19). As a whole, these findings suggest that the weak genetic signal observed on chromosome 17q is not specific to the PsA subphenotype.

A suggestive linkage was evidenced by Laval et al. (11) on chromosome 16q in AS, near marker D16S3091 located at 101 cM from the p-telomere (LOD=4.7, {lambda}=1.8). Notwithstanding the absence of linkage observed in our genome-screen, and considering the level of significance of this locus on the British linkage study, we further genotyped the marker D16S3091 in the extension study. We observed a trend in favour of linkage, but the NPL value did not reach the threshold of significance (NPL=1.7; LOD=0.63, P=0.04) despite a power higher to that of Laval et al. [336 affected relative pairs (Table 2) versus 255 affected sibling pairs]. Our result may thus appear as discordant to the previous report. In fact, in complex genetic disorders, linkage is often difficult to replicate. Genetic heterogeneity from a population to another (e.g. French versus British) may be explicative, but sample variations occurring in the same population may be sufficient to explain such discrepancies. As a result, our data do not conflict with the report by Laval et al. of a SpA susceptibility locus on chromosome 16q.


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  		Table 2. Characteristics of the multiplex SpA families included in the study
 
Differences in disease definition also may be questioned for explaining this apparent discordance. The linkage evidenced on the British study involved patients with AS according to the modified New York's criteria (20) in which an advanced radiological sacroiliitis is necessary for diagnosis, as discussed before. Owing to the diagnosis criteria chosen in our study, the patients included did not necessarily exhibit advanced radiological changes. When comparing the clinical characteristics of our patients (2,3) with those included in the previous AS linkage study (11), extra-articular manifestations such as psoriasis and IBD occurred in comparable percentage of patients (respectively, 16 versus 12% and 9 versus 5%). Sex ratio was also similar (1.33 versus 1.43), so was HLA-B27 carriage (97 versus 100%). The only noticeable difference was a higher percentage of patients with uveitis reported by Laval et al. (44%) when compared to our cohort of patients (29%). However, uveitis is known to be correlated to the disease duration (2) and after stratification for this parameter, prevalence of uveitis was comparable in both cohorts (44 versus 47% after more than 19 years of disease duration). Altogether, these findings suggest that in the French and British studies, similar patients were analyzed but at different stages of disease progression, our approach including patients at an earlier phase of the disease, before the constitution of a radiographic sacroiliitis. However, it cannot be ruled out that the chromosome 16q locus is related to more severe diseases. Such hypothesis is sustained by a quantitative trait locus linkage analysis performed in the British cohort of AS patients, which demonstrated an overlapping linkage with the Bath Ankylosing Spondylitis Functional Index (BASFI) and with the area linked to susceptibility (29).

The most striking finding of this study was obtained on chromosome 9q, where a positive linkage was obtained on 9q31–34 (NPL=4.87, LOD=5.15, P=0.00002 at 120 cM from the p-telomere). According to Lander and Kruglyak guidelines for evaluation of results from linkage analyses of complex diseases, this value is significant enough to conclude that a SpA susceptibility locus resides in this region. Thus, this study provides the confirmation of the presence of a non-MHC gene involved in SpA predisposition. Interestingly, Laval et al. (11) previously reported suggestive linkage for AS in a nearby region (two point LOD score=2.3, P=0.0006 for marker D9S1682 at 133 cM). Altogether, these data clearly indicate that a SpA susceptibility gene resides on the 9q31–34 region. The calculated {lambda}S value for this locus is 1.75 in our family set and 1.5 in the study reported by Laval et al. These values are consistent and suggest that the gene encoded at this locus is of moderate effect in SpA susceptibility, as expected for a complex genetic disorder.

Looking at the results pedigree-by-pedigree, we were able to detect an extended pedigree contributing in a large proportion to the positive result observed on chromosome 9 (Fig. 2). This family included 10 affected relatives until the 5th degree, all sharing a common haplotype on chromosome 9q. The affected members from this family were recruited through the same diagnosis criteria than the whole family set and did not differ on their clinical characteristics (data not shown). Nevertheless, we observed a high prevalence of HLA-B27 positivity not only among affected family members (100%) but also among the healthy spouses (Fig. 2). Such HLA-B27 prevalence potentially enhanced the disease expressivity among the kindreds explaining this large familial aggregation.

By examining the co-segregation of this haplotype with the disease status within the family, and by taking into account the HLA-B27 carrier status, we inferred a possible dominant model of inheritance with incomplete penetrance. This conclusion was supported by further parametric analyses that propose a dominant model with incomplete penetrances and a rare at risk allele.

As expected for non-parametric analyses performed in a complex genetic trait for a locus with a moderate effect, the confidence interval of the SpA locus mapping is rather large. Transmission disequilibrium tests using the available genotyping data failed to detect an association and did not allow us to refine the locus position (data not shown). Considering the haplotype shared by affected members of pedigree 110, an interval of at least 10 cM (between markers D9S1776 and D9S1682) can be proposed (Fig. 2).

Almost 150 genes have been mapped currently to the 9q31–34 interval. Of these, TLR4, TRAF1 and CD30L are involved in the regulation of the immune response and can be proposed as the most noticeable functional candidates. TLR4 is a well-characterized member of the Toll-like receptors that mediates the innate response to the lipopolysaccharide (LPS), a bacterial cell-wall component (30,31). TLR4 mediates a signalling cascade to the nucleus leading to the activation of NF-{kappa}b. Structural and functional homologies exist between TLR4 and NOD2/CARD15, which has been demonstrated to be involved in CD susceptibility (32,33). Moreover, bacterial components are suspected to be involved in the physiopathology of SpA, as in CD. Tumour necrosis factor receptor (TNFR)-associated factor 1 (TRAF1) is a component of the TNFR2 signalling complex involved in cell proliferation, cytokine production and cell death. TRAF1 regulates antigen-induced apoptosis of CD8+ lymphocytes in case of over-expression in transgenic mice (34). CD30L (TNFSFR8) is a cell membrane bound and a secreted molecule belonging to the TNF family of ligands, constitutively expressed by NK cells (35) and involved in NF-{kappa}B activation (36). Systematic analysis of these candidate genes as well as linkage disequilibrium mapping on 9q31–34 should permit further identification of the gene involved in SpA susceptibility. Such findings undoubtedly will be of great importance for a better understanding of the disease mechanisms and the development of new therapeutic approaches for treating SpA.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 RESULTS
 DISCUSSION
 MATERIALS AND METHODS
REFERENCES
 
Families and phenotype
Caucasian SpA families were recruited through the Groupe Français d'Etude Génétique des Spondylarthropathies (GFEGS). In order to limit the underlying genetic heterogeneity of the trait, we choose to perform linkage analyses only on French families. The study was approved by the relevant local ethics committee and informed consent was obtained from each participant. The diagnosis of SpA was made according to the classification criteria previously proposed by Amor (21) and/or according to the ESSG diagnosis criteria (22). Within the group of SpA, AS was diagnosed according to the modified New York criteria (20). Regarding extra-articular manifestations, the diagnosis of psoriasis required the presence of typical lesions and/or a clinical diagnosis established by a dermatologist. Anterior uveitis was retained after examination by an ophthalmologist. IBD diagnosis (including CD and ulcerative colitis) was based on endoscopic and histological examination of the gut. ReA was diagnosed according to the criteria published by Willkens (37). Finally, USpA was retained when SpA criteria were fulfilled, without AS, PsA, ReA or IBD.

Two independent panels of Caucasian multiplex families were consecutively analyzed in a two-stage strategy. The first family set (65 families including 151 affected relative pairs) was analyzed in a first step for a whole genome scan. The second family set (55 families including 185 affected relative pairs) was further investigated for an extension study focusing on genetic regions identified in the genome-wide screen. Families from both panels were recruited according to the same diagnostic criteria. A higher percentage of peripheral arthritis and back pain was observed among the affected members from the second set of families, presumably due to sample variations of the same population. Nevertheless, no statistical differences were evidenced regarding the clinical forms of the disease (P=0.6) (Table 3). Of the 120 recruited families, 80 were nuclear and 40 were extended pedigrees (Table 2). Among the nuclear families, both parents were genotyped in 72% of cases and one parent in 20%. In the case where parents where not genotyped, unaffected relatives were used to increase the information available for calculation of identity-by-descent in the majority of cases.


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  		Table 3. Clinical characteristics and clinical forms of the disease among patients included in the genome screen and in the extension study
 
The total number of affected pairs, without correction for independence, were, respectively, 151 and 185 in panels 1 (whole-genome screen) and 2 (extension study). Overall, a total of 348 SpA patients and 535 unaffected relatives were genotyped.

Microsatellite markers and maps
The genome-wide screen was performed using the 352 autosomal and 17 X-linked microsatellite-markers from the Applied Biosystems fluorescent-labelled human linkage mapping set (LMS) version 2.0. The inter-marker spacing was on an average 9.25 cM (range 6.86–11.24). The mean marker heterozygosity was 0.77. For the extension study, 22 microsatellite markers were genotyped (D5S2027, D5S471, D5S467, D5S2059, D5S1984, D5S2002, D5S2115, D9S1677, D9S1776, D9S1682, D9S290, D9S1826, D13S217, D13S171, D13S218, D13S263, D16S3091, D17S787, D17S944, D17S949, D17S785, D17S784). Microsatellite-markers were amplified by PCR (30 cycles) in multiplex amplifications on ABI 877 integrated thermal cyclers (Applied Biosystems) in 5 µl reactions containing 10 ng of genomic DNA, 2.5 mM MgCl2, 250 µM dNTPs (Amersham Bioscience), 0.3–1.2 µM of each pooled primer and 0.1 units of AmpliTaq-Gold DNA polymerase (Applied Biosystems). PCR products were denatured at 95°C for 1 min and loaded on a 5% polyacrylamide gel (36 cm well-to-read plate) in a ABI 377 DNA analyzer (Applied Biosystems). The fluorescent signal was analyzed for semi-automated fragment sizing by the GeneScan version 2.1 software (Applied Biosystems). Allele calling was then performed using Genotyper software, version 2.0 (Applied Biosystems). Control DNA (individual 1347-02 from the CEPH reference families) was systematically used as an internal standard. Two investigators determined each genotype independently to confirm their accuracy. Conflicting data were either resolved or discarded.

Mendelian segregation of alleles was checked using the Unknown software version 5.23. Marker ordering and inter-marker distances were computed using the CRI-MAP program (38). The resulting maps, obtained from the recombination events observed in our first set of 65 families (genome-wide analysis), were in accordance with published data (data not shown) and were used for further analyses.

Statistical analysis
As the mode of inheritance of SpA is unknown, genotyping data were analyzed using the non-parametric multipoint linkage (NPL) program of the GENEHUNTER package version 2.0 (39). The program was configured for large pedigrees and simultaneous scoring of all possible affected relative pairs. Linkages observed on the pooled data were considered as ‘suggestive’ (P<7.4.10–4) or ‘significant’ (P<2.2.10–5) according to published guidelines (18). Parametric linkage analyses were also performed using the GENEHUNTER package (39). In order to assess the best penetrances, the maximum LOD score was computed over multiple parameter values. Penetrances were incremented by a grid space of 0.05. The gene frequency was also varied. Using the ‘heterogeneity’ function of GENEHUNTER, LOD scores under heterogeneity were also calculated and {alpha} value was varied until the HLOD was maximized.


   ACKNOWLEDGEMENTS
 
We acknowledge the patients and their families who participated in this study and all the members of the GFEGS for their contribution in organizing the sample collection: Maria Antonietta D'Agostino, MD, Cochin Hospital, Paris, France; Bernard Amor, MD, Cochin Hospital, Paris, France; Jean-Marie Berthelot, MD, University Hospital, Nantes, France; Alain Saraux, MD, PhD, Cavale Blanche Hospital, Brest, France; Aleth Perdriger, MD, PhD, University Hospital, Rennes, France; Sandrine Guis, MD, Conception Hospital, Marseille, France; Xavier Puechal, MD, Le Mans Hospital, Le Mans, France; Jean-Françis Maillefert, MD, PhD, General Hospital, Dijon, France; Bernard Combe, MD, PhD, Lapeyronie Hospital, Montpellier, France; Pascal Claudepierre, MD, Henri Mondor Hospital, Créteil, France; Jean Sibilia, MD, PhD, Hautepierre Hospital, Strasbourg, France. We are grateful to M. Legrand, L. Cazes, L. Piouffre, C. Vaury, P. Pastureau, J.C. Baudoin, H. Bui, A. Martins, E. Tubacher and E. Poullier. We thank T. Tsuk of the Association Française des Spondylarthritiques for his contribution to the allele calling. We thank Leigh Pascoe for critically reading the manuscript. This study was supported by grants from Société Française de Rhumatologie, Programme Hospitalier de Recherche Clinique, Association de Recherche sur la Polyarthrite, Fond d'Etude et de Recherche de l'Assistance Publique, Association de Recherche Clinique en Rhumatologie, Institut National de la Santé et de la Recherche Médicale and the Pfizer-Pharmacia-Searle company.


   FOOTNOTES
 
* To whom correspondence should be addressed at: Centre d'Etude du Polymorphisme Humain/Fondation Jean-Dausset27, rue Juliette Dodu, 75010 Paris, France. Tel: +33 153725020; Fax: +33 153725058; Email: corinne.miceli{at}cephb.fr


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 RESULTS
 DISCUSSION
 MATERIALS AND METHODS
 REFERENCES
 

  1. Saraux, A., Guedes, C., Allain, J., Devauchelle, V., Valls, I., Lamour, A., Guillemin, F., Youinou, P. and Le Goff, P. (1999) Prevalence of rheumatoid arthritis and spondyloarthropathy in Brittany, France. Societe de Rhumatologie de l'Ouest. J. Rheumatol., 26, 2622–2627.[Web of Science][Medline]

  2. Said-Nahal, R., Miceli-Richard, C., Berthelot, J.M., Duche, A., Dernis-Labous, E., Le Blevec, G., Saraux, A., Perdriger, A., Guis, S., Claudepierre, P. et al. (2000) The familial form of spondylarthropathy: a clinical study of 115 multiplex families. Groupe Francais d'Etude Genetique des Spondylarthropathies. Arthritis Rheum., 43, 1356–1365.[CrossRef][Web of Science][Medline]

  3. Said-Nahal, R., Miceli-Richard, C., D'Agostino, M.A., Dernis-Labous, E., Berthelot, J.M., Duche, A., Le Blevec, G., Saraux, A., Perdriger, A., Guis, S. et al. (2001) Phenotypic diversity is not determined by independent genetic factors in familial spondylarthropathy. Arthritis Rheum., 45, 478–484.[CrossRef][Web of Science][Medline]

  4. Brewerton, D.A., Hart, F.D., Nicholls, A., Caffrey, M., James, D.C. and Sturrock, R.D. (1973) Ankylosing spondylitis and HL-A 27. Lancet, 1, 904–907.[Web of Science][Medline]

  5. Schlosstein, L., Terasaki, P.I., Bluestone, R. and Pearson, C.M. (1973) High association of an HL-A antigen, W27, with ankylosing spondylitis. N. Engl. J. Med., 288, 704–706.[Web of Science][Medline]

  6. Hammer, R.E., Maika, S.D., Richardson, J.A., Tang, J.P. and Taurog, J.D. (1990) Spontaneous inflammatory disease in transgenic rats expressing HLA-B27 and human beta 2m: an animal model of HLA-B27-associated human disorders. Cell, 63, 1099–1112.[CrossRef][Web of Science][Medline]

  7. Jarvinen, P. (1995) Occurrence of ankylosing spondylitis in a nationwide series of twins. Arthritis Rheum., 38, 381–383.[Medline]

  8. Brown, M.A., Kennedy, L.G., MacGregor, A.J., Darke, C., Duncan, E., Shatford, J.L., Taylor, A., Calin, A. and Wordsworth, P. (1997) Susceptibility to ankylosing spondylitis in twins: the role of genes, HLA, and the environment. Arthritis Rheum., 40, 1823–1828.[Web of Science][Medline]

  9. van der Linden, S.M., Valkenburg, H.A., de Jongh, B.M. and Cats, A. (1984) The risk of developing ankylosing spondylitis in HLA-B27 positive individuals. A comparison of relatives of spondylitis patients with the general population. Arthritis Rheum., 27, 361–368.[Web of Science][Medline]

  10. van der Linden, S.M. and Khan, M.A. (1984) The risk of ankylosing spondylitis in HLA-B27 positive individuals: a reappraisal. J. Rheumatol., 11, 727–728.[Web of Science][Medline]

  11. Laval, S.H., Timms, A., Edwards, S., Bradbury, L., Brophy, S., Milicic, A., Rubin, L., Siminovitch, K.A., Weeks, D.E., Calin, A. et al. (2001) Whole-genome screening in ankylosing spondylitis: evidence of non-MHC genetic-susceptibility loci. Am. J. Hum. Genet., 68, 918–926.[CrossRef][Web of Science][Medline]

  12. Breban, M., Said-Nahal, R., Hugot, J.P. and Miceli-Richard, C. (2003) Familial and genetic aspects of spondyloarthropathy. Rheum. Dis. Clin. North Am., 29, 575–594.[CrossRef][Web of Science][Medline]

  13. Brown, M.A., Edwards, S., Hoyle, E., Campbell, S., Laval, S., Daly, A.K., Pile, K.D., Calin, A., Ebringer, A., Weeks, D.E. and Wordsworth, B.P. (2000) Polymorphisms of the CYP2D6 gene increase susceptibility to ankylosing spondylitis. Hum. Mol. Genet., 9, 1563–1566.[Abstract/Free Full Text]

  14. Tsui, F.W., Tsui, H.W., Cheng, E.Y., Stone, M., Payne, U., Reveille, J.D., Shulman, M.J., Paterson, A.D. and Inman, R.D. (2003) Novel genetic markers in the 5'-flanking region of ANKH are associated with ankylosing spondylitis. Arthritis Rheum., 48, 791–797.[CrossRef][Web of Science][Medline]

  15. McGarry, F., Neilly, J., Anderson, N., Sturrock, R. and Field, M. (2001) A polymorphism within the interleukin 1 receptor antagonist (IL-1Ra) gene is associated with ankylosing spondylitis. Rheumatology (Oxford), 40, 1359–1364.

  16. Maksymowych, W.P., Reeve, J.P., Reveille, J.D., Akey, J.M., Buenviaje, H., O'Brien, L., Peloso, P.M., Thomson, G.T., Jin, L. and Russell, A.S. (2003) High-throughput single-nucleotide polymorphism analysis of the IL1RN locus in patients with ankylosing spondylitis by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. Arthritis Rheum., 48, 2011–2018.[CrossRef][Web of Science][Medline]

  17. Brown, M.A., Pile, K.D., Kennedy, L.G., Campbell, D., Andrew, L., March, R., Shatford, J.L., Weeks, D.E., Calin, A. and Wordsworth, B.P. (1998) A genome-wide screen for susceptibility loci in ankylosing spondylitis. Arthritis Rheum., 41, 588–595.[CrossRef][Web of Science][Medline]

  18. Lander, E. and Kruglyak, L. (1995) Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results. Nat. Genet., 11, 241–247.[CrossRef][Web of Science][Medline]

  19. Altmuller, J., Palmer, L.J., Fischer, G., Scherb, H. and Wjst, M. (2001) Genomewide scans of complex human diseases: true linkage is hard to find. Am. J. Hum. Genet., 69, 936–950.

  20. van der Linden, S., Valkenburg, H.A. and Cats, A. (1984c) Evaluation of diagnostic criteria for ankylosing spondylitis. A proposal for modification of the New York criteria. Arthritis Rheum., 27, 361–368.[Web of Science][Medline]

  21. Amor, B., Dougados, M. and Mijiyawa, M. (1990) Critère diagnostique des spondylarthropathies. Rev. Rhum. Mal. Ostéoartic., 57, 85–89.[Medline]

  22. Dougados, M., Van der Linden, S., Juhlin, R., Huitfeldt, B., Amor, B., Calin, A., Cats, A., Dijkmans, B., Olivieri, I., Pasero, G. et al. (1991) The European Spondylarthropathy Study Group preliminary criteria for the classification of spondylarthropathy. Arthritis Rheum., 34, 1218–1227.[Web of Science][Medline]

  23. Collantes, E., Veroz, R., Escudero, A., Munoz, E., Munoz, M.C., Cisnal, A. and Perez, V. (2000) Can some cases of ‘possible’ spondyloarthropathy be classified as ‘definite’ or ‘undifferentiated’ spondyloarthropathy? Value of criteria for spondyloarthropathies. Spanish Spondyloarthropathy Study Group. Joint Bone Spine, 67, 516–520.[CrossRef][Web of Science][Medline]

  24. Wordsworth, P. (1998) Genes in the spondyloarthropathies. Rheum. Dis. Clin. North Am., 24, 845–863.[CrossRef][Web of Science][Medline]

  25. Rioux, J.D., Daly, M.J., Silverberg, M.S., Lindblad, K., Steinhart, H., Cohen, Z., Delmonte, T., Kocher, K., Miller, K., Guschwan, S. et al. (2001) Genetic variation in the 5q31 cytokine gene cluster confers susceptibility to Crohn disease. Nat. Genet., 29, 223–228.[CrossRef][Web of Science][Medline]

  26. Tomfohrde, J., Silverman, A., Barnes, R., Fernandez-Vina, M.A., Young, M., Lory, D., Morris, L., Wuepper, K.D., Stastny, P., Menter, A. and Bowcock, A.M. (1994) Gene for familial psoriasis susceptibility mapped to the distal end of human chromosome 17q. Science, 264, 1141–1145.[Abstract/Free Full Text]

  27. Nair, R.P., Henseler, T., Jenisch, S., Stuart, P., Bichakjian, C.K., Lenk, W., Westphal, E., Guo, S.W., Christophers, E., Voorhees, J.J. and Elder, J.T. (1997) Evidence for two psoriasis susceptibility loci (HLA and 17q) and two novel candidate regions (16q and 20p) by genome-wide scan. Hum. Mol. Genet., 6, 1349–1356.[Abstract/Free Full Text]

  28. Enlund, F., Samuelsson, L., Enerback, C., Inerot, A., Wahlstrom, J., Yhr, M., Torinsson, A., Riley, J., Swanbeck, G. and Martinsson, T. (1999) Psoriasis susceptibility locus in chromosome region 3q21 identified in patients from southwest Sweden. Eur. J. Hum. Genet., 7, 783–790.[CrossRef][Web of Science][Medline]

  29. Brown, M.A., Brophy, S., Bradbury, L., Hamersma, J., Timms, A., Laval, S., Cardon, L., Calin, A. and Worsworth, P. (2003) Dentification of major loci controlling clinical manifestations of ankylosing spondylitis. Arthritis Rheum., 48, 2234–2239.[CrossRef][Web of Science][Medline]

  30. Poltorak, A., He, X., Smirnova, I., Liu, M.Y., Van Huffel, C., Du, X., Birdwell, D., Alejos, E., Silva, M., Galanos, C. et al. (1998) Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science, 282, 2085–2088.[Abstract/Free Full Text]

  31. Chow, J.C., Young, D.W., Golenbock, D.T., Christ, W.J. and Gusovsky, F. (1999) Toll-like receptor-4 mediates lipopolysaccharide-induced signal transduction. J. Biol. Chem., 274, 10689–10692.[Abstract/Free Full Text]

  32. Hugot, J.P., Chamaillard, M., Zouali, H., Lesage, S., Cezard, J.P., Belaiche, J., Almer, S., Tysk, C., O'Morain, C.A., Gassull, M. et al. (2001) Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease. Nature, 411, 599–603.[CrossRef][Medline]

  33. Ogura, Y., Bonen, D.K., Inohara, N., Nicolae, D.L., Chen, F.F., Ramos, R., Britton, H., Moran, T., Karaliuskas, R. and Duerr R.H. (2001) A frameshift mutation in NOD2 associated with susceptibility to Crohn's disease. Nature, 411, 603–606.[CrossRef][Medline]

  34. Speiser, D.E., Lee, S.Y., Wong, B., Arron, J., Santana, A., Kong, Y.Y., Ohashi, P.S. and Choi, Y. (1997) A regulatory role for TRAF1 in antigen-induced apoptosis of T cells. J. Exp. Med., 185, 1777–1783.[Abstract/Free Full Text]

  35. Kashii, Y., Giorda, R., Herberman, R.B., Whiteside, T.L. and Vujanovic, N.L. (1999) Constitutive expression and role of the TNF family ligands in apoptotic killing of tumor cells by human NK cells. J. Immunol., 163, 5358–5366.[Abstract/Free Full Text]

  36. Aizawa, S., Nakano, H., Ishida, T., Horie, R., Nagai, M., Ito, K., Yagita, H., Okumura, K., Inoue, J. and Watanabe, T. (1997) Tumor necrosis factor receptor-associated factor (TRAF) 5 and TRAF2 are involved in CD30-mediated NFkappaB activation. J. Biol. Chem., 272, 2042–2045.[Abstract/Free Full Text]

  37. Willkens, R.F., Arnett, F.C., Bitter, T., Calin, A., Fisher, L., Ford, D.K., Good, A.E. and Masi, A.T. (1981) Reiter's syndrome. Evaluation of preliminary criteria for definite disease. Arthritis Rheum., 24, 844–849.[Web of Science][Medline]

  38. Lander, E.S. and Green, P. (1987) Construction of multilocus genetic linkage maps in humans. Proc. Natl Acad. Sci. USA, 84, 2363–2367.[Abstract/Free Full Text]

  39. Kruglyak, L., Daly, M.J., Reeve-Daly, M.P. and Lander, E.S. (1996) Parametric and nonparametric linkage analysis: unified multipoint approach. Am. J. Hum. Genet., 58, 1347–1363.[Web of Science][Medline]


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