HLA-B27 Positive Ankylosing Spondylitis Patients Research Articles

Generation of induced pluripotent stem cells from an HLA-B27 positive ankylosing spondylitis patient with syndesmophyte formation

Human leukocyte antigen (HLA)-B27 is the genetic marker for ankylosing spondylitis (AS). Here, we generated induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells of a male AS patient carrying HLA-B27 with syndesmophyte formation by using the Sendai-virus delivery system. The resulting iPSCs had a normal karyotype, expressed pluripotent markers, and could differentiate into three germ layers. This cellular model will provide a platform for studying pathological mechanisms of new bone formation in HLA-B27 positive AS patients.

Syndesmophyte Growth in Ankylosing Spondylitis: from Laboratory to Bedside.

This study aims to review recent studies on risk factors for syndesmophyte growth in ankylosing spondylitis (AS) and on treatment effects. New genetic studies, including a genome-wide association study, provided only limited evidence of specific genetic associations with radiographic severity. Measures of inflammation, including vertebral osteitis and C-reactive protein level, were strongly associated with radiographic progression, while studies of adipokines had mixed results. Mesenchymal stem cells from HLA-B27 positive AS patients were found to promote vertebral ossification via a pathway of B27 misfolding, retinoic acid receptor-β activation, and increased bone alkaline phosphatase. Low vertebral trabecular bone density is associated with syndesmophyte growth, with reciprocal effects when bridged. Several observational studies suggested radiographic severity was reduced by treatment with tumor necrosis factor inhibitors, particularly when longer than 2years. Syndesmophyte development in AS is the result of a complex, incompletely understood, interplay of inflammatory and mechanical factors.

Genetic effects of B3GNT2 on ankylosing spondylitis susceptibility and clinical manifestations in Taiwanese

The intergenic SNP rs10865331 at 2p15 was identified as a major risk factor for ankylosing spondylitis (AS) susceptibility in genome-wide association studies (GWAS). B3GNT2 gene regulates polylactosamine synthesis is potentially functionally relevant to AS disease development. We investigated whether SNP rs10865331 and two B3GNT2 SNPs (rs11900673 and rs1136151) are associated with AS susceptibility and disease severity in Taiwanese. Distributions of genotypes, alleles, and haplotypes of three SNPs were compared between 1,472 AS patients and 2,117 healthy blood donors and among AS patients stratified by clinical characteristics. The intergenic SNP rs10865331 was significantly associated with AS (PFDR=1.02E-05) in Taiwanese. In AS patients stratified by positivity of HLA-B27 and syndesmophyte formation, all three B3GNT2 locus SNPs (rs11900673, rs1136151, and rs10865331) were significantly associated with syndesmophyte formation among HLA-B27 positive AS patients. Haplotype analyses revealed that the "CTA" (rs11900673C/rs1136151T/rs10865331A) haplotype was significantly associated with AS susceptibility (Padj=0.0177) and syndesmophyte formation (Padj=0.016) in HLA-B27 positive patients. In contrast, "TCG" (rs11900673T/rs1136151C/rs10865331G) haplotype showed protection against AS development (Padj=0.0005 for HLA-B27 positive and Padj=0.004 for HLA-B27 negative, respectively) and syndesmophyte formation (Padj=0.0017) in HLA-B27 positive patients. Furthermore, B3GNT2 mRNA expressions were negatively associated with erythrocyte sedimentation rate (ESR, P=0.0103), C-reactive protein (CRP, P=0.0353), Bath ankylosing spondylitis functional index (BASFI, P=0.0171), and syndesmophyte formation (P=0.0148). Our data suggest that B3GNT2 gene may contribute to AS development and affect AS severity by interacting with HLA-B27 in Taiwanese.

Choroidal thickness changes determined by EDI-OCT on acute anterior uveitis in patients with HLA-B27-positive ankylosing spondylitis.

To evaluate the choroidal thickness on acute anterior uveitis in patients with HLA-B27-positive ankylosing spondylitis. In this study, 32 eyes of 16 HLA-B27 positive AS patients with anterior uveitis and age-matched 19 eyes for control group were analyzed between January 2014 and April 2015. Assessment criteria were uveitis activity, visual acuity, flare existence, subfoveal choroidal thickness and central macular thickness measurements. The mean subfoveal choroidal thicknesses in affected eye group (Group 1), unaffected eye group (Group 2) and control group (Group 3) were 348.31±72.7, 301.12±49.2 and 318.0±74.3, respectively, in active periods. (p=0.04 between Group 1 and Group 2, p=0.234 between Group 1 and Group 3) The mean central macular thicknesses of Group 1, Group 2 and Group 3 were 268.50±16.5, 267.31±16.3 and 249.7±30.5, respectively, in active periods. (p=0.84 between Group 1 and Group 2, p=0.029 between Group 1 and Group 3). However, in convalescence period, the mean subfoveal choroidal thicknesses of Group 1, Group 2 and Group 3 were 322.40±48.5, 300.75±47.7 and 318.0±74.3, respectively. (p=0.22 between Group 1 and Group 2, p=0.854 between Group 1 and Group 3) The mean central macular thicknesses of Group 1, Group 2 and Group 3 were 269.75±21.9, 256.62±21.5 and 249.7±30.5, respectively. (p=0.09 between Group 1 and Group 2, p=0.03 between Group 1 and Group 3). In HLA-B27 positive ankylosing spondylitis patients with anterior uveitis, the choroidal thicknesses of the affected eyes were found as thicker than fellow unaffected one or control eyes in active period. The central macular thicknesses are not affected on both active and convalescent period.

Diagnostic value of anti-MCV antibodies for ankylosing spondylitis

Objective To investigate the diagnostic value of anti-mutated citrullinated vimentin（anti-MCV） and anti-cyclic citrullinated peptide（anti-CCP） antibodies for ankylosing spondylitis (AS), and to establish a new diagnostic method of AS based on anti-citrullinated protein/peptide antibodies（ACPAs） detection. Methods Totally 121 AS patients(AS group), 97 rheumatoid arthritis（RA） patients(RA group)and 103 healthy people(control group) were enrolled for the detection of serum levels of anti-MCV and anti-CCP antibodies using ELISA method as a diagnostic test. HLA-B27 in AS patients was tested by flow cytometry, and RF-IgM in RA and AS patients was detected by immune rate nephelometry. Receiver operating characteristics (ROC) curve was used to analyze the diagnostic value and to determine the cut-off value. Anti-MCV and anti-CCP antibodies among each group were compared by Mann-Whitney U test, Chi-square test or Fisher′s exact test was used to compare positive rate. Results The anti-MCV levels in AS group ［11.60 (12.36-25.83) U/ml］were significantly higher than control group ［11.60(8.41-13.54）U/ml ］,U=2 413,P<0.001, while the levels of anti-CCP had no difference between the two groups ［AS group 6.22 （4.30-8.07）U/ml］, and control group ［6.01(3.77-7.58)U/ml］, respectively; U=5 421,P=0.094. The calculated area under the ROC curve of anti-MCV was 0.806,and 14.67 U/ml was the optimal cut-off value with sensitivity of 0.645 and specificity of 0.942. In both HLA-B27 positive and negative AS patients, anti-MCV antibodies levels and positive rate were significantly higher than control group using new cut-off value above （U=133.5, P=0.001; U=2 279.5,P<0.001). Sensitivity of the combination detection of anti-MCV and anti-CCP (MCV+CCP－) or RF-IgM (MCV+RF－) were 60.3% (73/121), 62.8% (76/121) and specificity were 89.7% (87/97), 90.7% (88/97) respectively in differential diagnosis of AS and RA, which were significantly higher than anti-MCV detection alone in specificity (16.5%,16/97). Conclusions Anti-MCV could be a new biomarker for the diagnosis of AS. With high sensitivity and specificity, anti-MCV has an equal diagnostic efficiency in HLA-B27 positive and negative AS patients using our new cut-off value. Additionally， the combination detection of anti-MCV and anti-CCP or RF-IgM could be an effective method for differential diagnosis of AS and RA.（Chin J Lab Med,2014,37: 455-458） Key words: Spondylitis,ankylosing; Peptides,cyclic; Vimentin; Mutation; Rheumatoid factor

OP0279 The Unfolded Protein Response Genes in Synovium and Blood Mononuclear Cells of HLA-B27 Positive Ankylosing Spondylitis Patients is not Increased Compared to Other Arthritis Patients and Healthy Controls

BackgroundThe HLA-B27 heavy chain is prone to misfolding. Misfolded proteins give rise to endoplasmic reticulum stress and activation of the Unfolded Protein Response (UPR). The UPR is strongly activated in...

SAT0289 Mri assessment of spinal involvement in psoriatic arthritis: Extent of disease relates to HLA-B27

BackgroundPsoriatic Arthritis (PsA) is a heterogeneous disease with a variable, sometimes evolving clinical phenotype. An estimated 25-70% of patients may have spinal involvement which varies from largely asymptomatic to severe...

Genetic Polymorphisms of Stromal Interaction Molecule 1 Associated with the Erythrocyte Sedimentation Rate and C-Reactive Protein in HLA-B27 Positive Ankylosing Spondylitis Patients

Ankylosing spondylitis (AS) is a chronic inflammation of the sacroiliac joints, spine and peripheral joints. The development of ankylosing spondylitis is still unclear. Genetics factors such as human leukocyte antigen HLA-B27 and ERAP1 have been widely reported to associate to AS susceptibility. In this study, we enrolled 361 AS patients and selected four tagging single nucleotides polymorphisms (tSNPs) at STIM1 gene. The correlation between STIM1 genetic polymorphisms and AS activity index (BASDAI, BASFI, BAS-G) as well as laboratory parameters of inflammation (erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP)) were tested. Our results indicated that HLA-B27 positive AS patients who are carrying the minor allele homozygous G/G genotype of SNP rs3750996 significantly associated with a higher level of ESR in serum. Furthermore, rs3750996/rs3750994 pairwise allele analysis indicated that G-C haplotypes also significantly correlated with higher level of ESR as well as CRP. These findings provide a better understanding of STIM1 genetic contribution to the pathogenesis of AS.

Correlation of HLA B27 subtypes with clinical features of ankylosing spondylitis

The aim of the present study was to identify the B*27 subtypes associated with ankylosing spondylitis (AS) in our population and correlate them with clinical features of AS. Whole blood samples were collected from 81 HLA-B27 positive AS patients and 29 controls (asymptomatic healthy unrelated individuals) positive for HLA-B27. Clinical details of the patients were recorded which included history of inflammatory back pain, sacroiliitis, spine involvement, enthesitis, peripheral arthritis and uveitis. HLA-B27 subtypes were detected using commercially available techniques. Fisher's exact test was used for statistical analysis. The subtypes observed in AS patients were B*2705 (67.9%, 55/81), B*2704 (28.4%, 23/81), B*2707 (2/81) and B*2702 (1/81). Subtypes in the controls were B*2705 (62.07%, 18/29), B*2707 (27.59%, 8/29) and B*2704 (10.34%, 3/29). Uveitis was observed more in B*2704-positive AS patients (34.78%, 8/23) compared to B*2705-positive AS patients (16.36%, 9/55). However, the difference was not statistically significant (P = 0.130). No major differences were found between B*2705 and B*2704 for other clinical features. B*2705 was the main subtype observed in both patient and control groups. Frequency of B*2704 was more in AS patients compared to controls. Occurrence of AS-associated uveitis was more often in B*2704-positive AS patients compared to B*2705-positive ones.

HLA Haplotype A33-B58-Cw10 May Modulate Radiographic Development of Bamboo Spine in Taiwanese Patients with Primary Ankylosing Spondylitis

Objective: To evaluate the possible relationship between HLA alleles and bony ankylosis of the spine (bamboo spine) in Taiwanese patients with ankylosing spondylitis (AS).Methods: A small cohort of HLA-B27 positive AS patients was conducted to analyze the effects of alleles and haplotypes on the development of bamboo spine. DNA typing of HLA class I and class II genes were performed by SSP method on primary ankylosing spondylitis patients with bamboo spine (n= 84) and spinal enthesopathy controls (n= 228). Odds ratios with 95% confidence intervals andPvalue were estimated. Determination of the most probable HLA haplotypes on all patients were constructed by comparison of the alleles carried by each patient with the HLA haplotype database established in Taiwanese population studies using homozygosity approach [1] and by expectation-maximum algorithm [2].Results: Allele frequencies of HLA A33, B58, Cw10, DR4, DR17 and DQ2 were significantly lower in bamboo patients as compared to non-bamboo controls. In contrast, allele frequencies of A24, B54, Cw15, DR11 and DR14 were significantly higher in bamboo patients. Less remarkably, high frequencies of B39, B51, Cw1 and Cw2 alleles were also noted in bamboo patients. Considering linkage disequilibria of alleles in haplotypes, HLA-A11-B27-Cw12 was the most common haplotype in both bamboo and non-bamboo groups (95.23% vs. 91.22%, respectively,P= 0.238). Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 were significantly lower in bamboo patients as compared to those in controls (P&lt; 0.001,P= 0.001,P= 0.002, respectively).Conclusion: Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 showed a strong association with bamboo spine in Taiwanese AS patients. Detection of such haplotypes might be a great aid in the management of patients with the disease.

HLA haplotype A33-B58-Cw10 may modulate radiographic development of bamboo spine in Taiwanese patients with primary ankylosing spondylitis.

Objective: To evaluate the possible relationship between HLA alleles and bony ankylosis of the spine (bamboo spine) in Taiwanese patients with ankylosing spondylitis (AS).Methods: A small cohort of HLA-B27 positive AS patients was conducted to analyze the effects of alleles and haplotypes on the development of bamboo spine. DNA typing of HLA class I and class II genes were performed by SSP method on primary ankylosing spondylitis patients with bamboo spine (n = 84) and spinal enthesopathy controls (n = 228). Odds ratios with 95% confidence intervals and P value were estimated. Determination of the most probable HLA haplotypes on all patients were constructed by comparison of the alleles carried by each patient with the HLA haplotype database established in Taiwanese population studies using homozygosity approach [1] and by expectation-maximum algorithm [2].Results: Allele frequencies of HLA A33, B58, Cw10, DR4, DR17 and DQ2 were significantly lower in bamboo patients as compared to non-bamboo controls. In contrast, allele frequencies of A24, B54, Cw15, DR11 and DR14 were significantly higher in bamboo patients. Less remarkably, high frequencies of B39, B51, Cw1 and Cw2 alleles were also noted in bamboo patients. Considering linkage disequilibria of alleles in haplotypes, HLA-A11-B27-Cw12 was the most common haplotype in both bamboo and non-bamboo groups (95.23% vs. 91.22%, respectively, P = 0.238). Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 were significantly lower in bamboo patients as compared to those in controls (P < 0.001, P = 0.001, P = 0.002, respectively).Conclusion: Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 showed a strong association with bamboo spine in Taiwanese AS patients. Detection of such haplotypes might be a great aid in the management of patients with the disease.

A polymorphism within the Transforming Growth Factor β1 gene is associated with ankylosing spondylitis (AS)

Genetic factors that predispose individuals to ankylosing spondylitis (AS) are not fully understood. Axial and sacroiliac joint fibrosis are characteristic of AS and the presence of TGFβ1 mRNA in AS sacroiliac joints raised the possibility that this cytokine might be implicated in this fibrosis. We have therefore examined a group of HLA B27 positive AS patients to investigate whether they could be prone to fibrosis based on overproduction of TGFβ1.

Recognition of B cells epitopes of the Klebsiella pneumoniae GroEL-like protein by HLA-B27 positive subjects

The presence of antibodies against antigens of K. pneumoniae in HLA-B27 positive patients with ankylosing spondylitis (AS), has been well documented. We have previously reported that sera from HLA-B27 positive subjects react with the K. pneumoniae GroEL-like protein (HSP60Kp) and have higher titers than HLA-B27 negative individuals. We cloned the gene that codes for this protein, determined hydrophilic regions by computer analysis of the predicted amino acid sequence and found that residues 389–397, 360–368 and 282–290, were possible B cell epitopes. To test this prediction, and to determine if the HLA-B27 positive and negative AS patients recognize the same or different epitopes, we truncated thehsp60Kp gene, from the 3 terminal nucleotide, to obtain fragments having or not the predicted epitopes. Four polypeptides of 40, 37, 30 and 18 kDa were obtained and analysed, by ELISA and inhibition of ELISA, for their reactivity with IgG antibodies from three high responders HLA-B27 positive AS patients and three HLA-B27 negative subjects who recognized the rHSP60Kp. Sera from both HLA-B27 positive and negative subjects reacted equally well with rHSP60Kp or with the 40 and 37 kDa peptides, which do not have residues 389–397 and 360–368, respectively, but reactivity was lost with the 30 kDa peptide, which also lacks residues 282–290. Contrary to what we expected, antibodies from HLA-B27 negative and positive individuals recognized the same epitope of the HSP60Kp. Our results indicate that the important epitope for B cells could be the 282–290 region and that the contribution of the two other predicted regions is minimal. We also conclude that the differences in response to the HSP60Kp in HLA-B27 positive AS patients and HLA-B27 negative individuals is not qualitative, but only quantitative.

HLA class I associations of ankylosing spondylitis in the white population in the United Kingdom.

To investigate the HLA class I associations of ankylosing spondylitis (AS) in the white population, with particular reference to HLA-B27 subtypes. HLA-B27 and -B60 typing was performed in 284 white patients with AS. Allele frequencies of HLA-B27 and HLA-B60 from 5926 white bone marrow donors were used for comparison. HLA-B27 subtyping was performed by single strand conformation polymorphism (SSCP) in all HLA-B27 positive AS patients, and 154 HLA-B27 positive ethnically matched blood donors. The strong association of HLA-B27 and AS was confirmed (odds ratio (OR) 171, 95% confidence interval (CI) 135 to 218; p < 10(-99)). The association of HLA-B60 with AS was confirmed in HLA-B27 positive cases (OR 3.6, 95% CI 2.1 to 6.3; p < 5 x 10(-5)), and a similar association was demonstrated in HLA-B27 negative AS (OR 3.5, 95% CI 1.1 to 11.4; p < 0.05). No significant difference was observed in the frequencies of HLA-B27 allelic subtypes in patients and controls (HLA-B*2702, three of 172 patients v five of 154 controls; HLA-B*2705, 169 of 172 patients v 147 of 154 controls; HLA-B*2708, none of 172 patients v two of 154 controls), and no novel HLA-B27 alleles were detected. HLA-B27 and -B60 are associated with susceptibility to AS, but differences in HLA-B27 subtype do not affect susceptibility to AS in this white population.

The phagocyte oxidative metabolism function in ankylosing spondylitis.

The phagocyte oxydative metabolism function was measured using chemiluminescence in microamounts of whole blood in 15 ankylosing spondylitis (AS) patients (10 were B27 positive), and in 17 controls. It was obtained from cells at rest, and following stimulation (latex, zymosan, fMLP), with luminol and lucigenin as amplifiers. The maximal light intensity was significantly higher (P less than 0.01) in AS compared to the controls in resting cells as well as in those after stimulation. There was no difference between HLA-B27 positive or negative AS patients. The increase in oxidative metabolism of the phagocyte system in AS was more evident in the luminol dependent assay, suggesting an activation of the myeloperoxydase system.

Rabbit antisera to enterobacteriaceae isolated from HLA-B27 positive patients with ankylosing spondylitis (AS) are lytic for the mononuclear cells of AS patients.

Vaccines prepared from Gram-negative bacteria isolated from the stools of HLA-B27 positive AS patients were used to immunize rabbits. Three of the sera obtained were lytic in vitro for the mononuclear cells of HLA-B27 positive AS patients. One of these sera discriminated between AS patients and healthy HLA-B27 positive individuals. Cytolysis was determined in an automated, non-radioactive assay based on the release of carboxyfluorescein diacetate and the incorporation of propidium iodide.

Possible ankylosing spondylitis in acute anterior uveitis.

HLA-B27 typing of 103 acute anterior uveitis (AAU) patients showed that 49 (48%) were positive for this antigen. Of these HLA-B27 positive AAU patients, 27 (55%) had ankylosing spondylitis (AS) according to the New York criteria. The 22 HLA-B27 positive AAU patients without AS, however, also showed symptoms and signs of AS. Seventeen (77%) of these 22 HLA-B27 positive AS negative AAU patients fulfilled a set of criteria for classification as possible AS. In the group of 54 HLA-B27 negative AAU patients, only two patients (4%) had AS. Of the remaining 52 patients only seven (13%) had possible AS. Of the 21 patients having posterior uveitis (PU) none was HLA-B27 positive nor met the criteria for AS or possible AS. These findings indicate a strong relationship between AAU, AS or possible AS and HLA-B27 and suggest that HLA-B27 typing in AAU patients is important for early diagnosis of AS.