Análise da frequência de uveítes em pacientes com espondiloartrites, suas complicações e associação com parâmetros clínicos da doença / Analysis of the frequency of uveitis in spondyloarthritis patients and associations with clinical parameters of the disease

Resumo Objetivo: A uveíte anterior aguda é a principal manifestação extra-articular na espondiloartrite. O objetivo deste estudo foi analisar se a presença da uveíte se associa com diferentes manifestações clínicas, laboratoriais, radiológicas e a terapêutica nos pacientes com espondiloartrite. Métodos: Estudo observacional retrospectivo realizado com 153 pacientes portadores de espondiloartrite atendidos no período de 1997 a 2017 na Grande Florianópolis, Brasil. Foram analisados dados demográficos, laboratoriais, clínicos e do tratamento de pacientes com espondiloartrite em relação a presença ou não de uveíte. Resultados: A uveíte foi encontrada em 26,8% dos pacientes. A presença de complicações foi rara, ocorrendo catarata em somente quatro pacientes e glaucoma em dois deles. Foi observada uma tendência a maior frequência de uveíte anterior aguda no sexo masculino (p=0,06), nos pacientes com história familiar (p=0,19) e HLA-B27 positivos (p=0,14). Pacientes com espondiloartrite e uveíte mais frequentemente usavam anti-TNF (p=0,04) e apresentavam sacroiliite em exames de imagem (p=0,02). Não observou-se associação entre a uveíte e o acometimento cardiovascular (p=0,44), cutâneo (p=0,13) ou gastrointestinal (p=0,10). Conclusão: A uveíte que ocorre em pacientes com espondiloartrite é comum, tem predomínio no sexo masculino e é mais frequente em pacientes com HLA-B27 positivo. O uso de imunobiológicos como o anti-TNF é frequente nos pacientes com uveíte.

Abstract Objective: Acute anterior uveitis (AAU) is the most common extra-articular manifestation of spondyloarthritis. The aim of this study is to analyze if the presence of uveitis is associated with a diferent clinical manifestation, laboratorial, radiological and therapetiuc among spondyloarthritis patients. Methods: This was a observational retrospective study with 153 patients with spondyloarthritis attended in the period from 1997 to 2017 in Florianopolis, Brazil. It was analyzed demografical, laboratorial, clinical and therapeutic data in spondyloarthritis patients with or without uveitis. Results: 26,8% of the patients with spondyloarthritis presented uveitis. The presence of complications was rare, with cataract occurring in only four patients and glaucoma in two of them. A higher frequency of acute anterior uveitis in males (p = 0.06) was observed in patients with a family history (p = 0.19) and HLA-B27 positive (p = 0.14). Patients with spondyloarthritis and uveitis more frequently used anti-TNF (p = 0.04) and presented sacroiliitis on imaging tests (p = 0.02). There was no association between uveitis and cardiovascular (p = 0.44), cutaneous (p = 0.13) or gastrointestinal involvement (p = 0.10). Conclusion: Uveitis in patients with spondylarthritis is common, predominantly in males, and more frequently in HLA-B27 positive patients. The use of immunobiological agents such as anti-TNF is common in patients with uveitis.

Flow Cytometric Human Leukocyte Antigen-B27 Typing with Stored Samples for Batch Testing

BACKGROUND: Flow cytometry (FC) HLA-B27 typing is still used extensively for the diagnosis of spondyloarthropathies. If patient blood samples are stored for a prolonged duration, this testing can be performed in a batch manner, and in-house cellular controls could easily be procured. In this study, we investigated various methods of storing patient blood samples. METHODS: We compared four storage methods: three methods of analyzing lymphocytes (whole blood stored at room temperature, frozen mononuclear cells, and frozen white blood cells [WBCs] after lysing red blood cells [RBCs]), and one method using frozen platelets (FPLT). We used three ratios associated with mean fluorescence intensities (MFI) for HLAB27 assignment: the B27 MFI ratio (sample/control) for HLA-B27 fluorescein-5-isothiocyanate (FITC); the B7 MFI ratio for HLA-B7 phycoerythrin (PE); and the ratio of these two ratios, B7/B27 ratio. RESULTS: Comparing the B27 MFI ratios of each storage method for the HLA-B27+ samples and the B7/B27 ratios for the HLA-B7+ samples revealed that FPLT was the best of the four methods. FPLT had a sensitivity of 100% and a specificity of 99.3% for HLA-B27 assignment in DNA-typed samples (N=164) when the two criteria, namely, B27 MFI ratio >4.0 and B7/B27 ratio <1.5, were used. CONCLUSIONS: The FPLT method was found to offer a simple, economical, and accurate method of FC HLA-B27 typing by using stored patient samples. If stored samples are used, this method has the potential to replace the standard FC typing method when used in combination with a complementary DNA-based method.

Flow Cytometric Human Leukocyte Antigen-B27 Typing with Stored Samples for Batch Testing

BACKGROUND: Flow cytometry (FC) HLA-B27 typing is still used extensively for the diagnosis of spondyloarthropathies. If patient blood samples are stored for a prolonged duration, this testing can be performed in a batch manner, and in-house cellular controls could easily be procured. In this study, we investigated various methods of storing patient blood samples. METHODS: We compared four storage methods: three methods of analyzing lymphocytes (whole blood stored at room temperature, frozen mononuclear cells, and frozen white blood cells [WBCs] after lysing red blood cells [RBCs]), and one method using frozen platelets (FPLT). We used three ratios associated with mean fluorescence intensities (MFI) for HLAB27 assignment: the B27 MFI ratio (sample/control) for HLA-B27 fluorescein-5-isothiocyanate (FITC); the B7 MFI ratio for HLA-B7 phycoerythrin (PE); and the ratio of these two ratios, B7/B27 ratio. RESULTS: Comparing the B27 MFI ratios of each storage method for the HLA-B27+ samples and the B7/B27 ratios for the HLA-B7+ samples revealed that FPLT was the best of the four methods. FPLT had a sensitivity of 100% and a specificity of 99.3% for HLA-B27 assignment in DNA-typed samples (N=164) when the two criteria, namely, B27 MFI ratio >4.0 and B7/B27 ratio <1.5, were used. CONCLUSIONS: The FPLT method was found to offer a simple, economical, and accurate method of FC HLA-B27 typing by using stored patient samples. If stored samples are used, this method has the potential to replace the standard FC typing method when used in combination with a complementary DNA-based method.

Utility of In-House PCR for HLA-B27 Typing: Comparison of Concordance Rate between PCR Kit and In-House PCR / 대한진단검사의학회지

BACKGROUND: Commercial kits of PCR method are widely used in HLA-B27 typing; however, their cost is relatively high. In this study, we evaluated the utility of an in-house PCR method by comparing it with that of a commercial kit. METHODS: HLA-B27 typing was done in 188 patients by using two PCR methods, Absolute(TM) HLAB27 PCR kit (Biosewoom, Korea) and an in-house PCR method. The primers used in the in-house method were prepared by Bioneer (Korea). Both PCR tests were done by Gene Amp PCR System 9600 (Perkin-Elmer Centus Corp., USA). RESULTS: The commercial kit and in-house PCR showed 100% concordance rate with each other in HLA-B27 typing. Of 188 patients tested 72 (38.3%) were positive and 116 (61.7%) were negative by the both tests. Of 62 patients with ankylosing spondylitis, 50 were positive (80.7%). CONCLUSIONS: The in-house PCR is a reliable and cost-effective method and can replace or supplement commercial kits for HLA-B27 typing.

HLA-B27 Subtypes in Korean Patients with Ankylosing Spondylitis / 대한진단검사의학회지

BACKGROUND: HLA-B27 is strongly associated with ankylosing spondylitis (AS), and its subtypes differ in their ethnic distribution. Studies worldwide have shown that B*2701, B*2702, B*2704, B*2705, B*2707, B*2708, B*2714, B*2715, and B*2719 are AS-predisposing subtypes, whereas B*2706 and B*2709 are reported to be negatively associated with AS. The aim of this study was to investigate HLA-B27 polymorphism and clinical features according to subtypes in Korean patients with AS. METHODS: Two hundred thirty samples from patients with impression of AS were analyzed by polymerase chain reaction using a sequence-specific primers (PCR-SSP) method. Pel-Freez SSP Unitray HLA-B*27 kit (Dynal Biotech, USA) including 16 primers was used to define HLA-B27 subtypes from B*2701 to B*2735. RESULTS: Among 230 samples from patients with impression of AS, 171 were HLA-B27 positive, and among 160 patients diagnosed as AS, 154 (96.3%) were HLA-B27 positive, while 17 patients not diagnosed as AS were HLA-B27 positive. Among 154 HLA-B27 positive patients with AS, 142 (92.2%) were typed as B*2705 and 9 (5.8%) were typed as B*2704. Three cases (1.9%) could be interpreted only variously because of their HLA-B27 homogeneous alleles. Between B*2705 and B*2704, no specific HLA-B27 subtype appeared to contribute to AS susceptibility (P=0.60). Difference in clinical features between B*2705 and B*2704 could not be found in this study (P>0.05). CONCLUSIONS: This study verified that HLA-B27 (96.3%) is strongly associated with AS and identified that the major subtypes of HLA-B27 positive patients with AS in Korea are B*2705 (92.2%) and B*2704 (5.8%).

Reiter's disease [case report]

The authors describe a single case of classical Reiter's disease

Role of HLA-B27 in high grade atrioventricular block and sick sinus syndrome

There has been an association between HLA-B27 and cardiovascular affection. Furthermore, a significantly increased prevalence of HLA-B27 was found in men with pacemakers who had no clinical or radiological signs of HLA-B27 associated disease. This study was planned to obtain more insight into the association between HLA-B27 and high grade AV block [HG-AVB] and sick sinus syndrome [SSS], and the possible role of HLA-B27 in causing this block. HLA tissue typing was preformed on 200 normal Egyptians [control] and 75 Egyptian pts who had permanent pacemakers implanted for HG-AVB and SSS who had no clinical or radiological signs of HLA-B27 associated disease. The pacemaker pts were classified into 3 groups according to the aetiology of AVB and SSS, group 1 included 5 pts with acquired HG-AVB, group II included 10 pts with congenital complete heart block and group III included 60 pts with idiopathic HG-AVB or SSS. HLA-B27 was present in 5/200 [2.5%] normal healthy Egyptians, in 6/60 [10%] pts with idiopathic HG-AVB and SSS [group III]. However, HLA-B27 was negative in both groups I and II. There was a statistically significant higher prevalence of HLA-B27 in pts with idiopathic HG-AVB/SSS than in both normal Egyptians and pts with acquired or congenital AVB [p < 0.05]. HLA-B27 might have a role in the aetiology of idiopathic AVB however, further research is needed to obtain more insight into its role and the role of other factors in idiopathic AVB