Regulatory function of CD4+CD25++ T cells in patients with myasthenia gravis is associated with phenotypic changes and STAT5 signaling:1,25-Dihydroxyvitamin D3 modulates the suppressor activity

Regulatory T cells were investigated in early-onset (EO) and late-onset (LO) myasthenia gravis patients with anti-acetylcholine receptor antibody (AChR-MG). Alterations in PD-1 and PD-L1 on CD4(+)CD25(++) (Treg) and responder T cells (Tresp, CD4(+)CD25(-)) were observed in LOMG patients. GITR was decreased on CD4(+)CD25(++) of all patients. Decrease of FOXP3 was associated with lower phosphorylation of STAT5.1,25-dihydroxyvitamin D3 (VitD3) increased suppression in co-culture with a stronger effect in patients by acting possibly both on cell groups. Changes in surface molecules and intracellular pathways contribute to the defects of Treg in non-thymomatous AChR-MG and VitD3 can have modulatory effects. (C) 2015 Elsevier B.V. All rights reserved

Relation of HLA-DRB1 to IgG4 autoantibody and cytokine production in muscle-specific tyrosine kinase myasthenia gravis (MuSK-MG)

A small subset of myasthenia gravis (MG) patients develop autoantibodies against muscle-specific kinase (MuSK), which are predominantly of the immunoglobulin (Ig)G4 isotype. MuSK-MG is strongly associated with HLA-DRB1*14, HLA-DRB1*16 and HLA-DQB1*05. In this study, the possible effects of these HLA associations on MuSK IgG autoantibody or cytokine production were investigated. Samples from 80 MG patients with MuSK antibodies were studied. The disease-associated HLA types were screened in the DNA samples. The IgG1, IgG2, IgG3 and IgG4 titres of the MuSK antibodies and the levels of interleukin (IL)-4, IL-6, IL-17A and IL-10 were measured in the sera. Comparisons were made among the groups with or without HLA-DRB1*14, HLA-DRB1*16 or HLA-DQB1*05. The IgG4 titres of the MuSK antibodies were higher than those of the IgG1, IgG2 and IgG3 isotypes among the whole group of patients. DRB1*14 (+) DRB1*16 (–) patients had higher levels of IgG4 antibodies than those of DRB1*14 (–) DRB1*16 (+) patients. DRB1*14 (+) DRB1*16 (+) patients also had higher levels of IgG4 antibodies than those of DRB1*14 (–) DRB1*16 (+) and DRB1*14 (–) DRB1*16 (–) patients. Higher IL-10 and lower IL-17A levels were measured in DRB1*14 (+) DRB1*16 (–) patients than in DRB1*14 (–) DRB1*16 (–) patients. The higher IgG4 titres of MuSK autoantibodies in patients carrying HLA-DRB1*14 than those in the other patients suggest a role for HLA in the production of the antibodies. The differences in IL-10 and IL-17A support the role of DRB1 in the etiopathogenesis of this autoimmune response.Istanbul Universit

Turkish version of the motor function measure scale (MFM-32) for neuromuscular diseases: A cross-cultural adaptation, reliability, and validity study

WOS: 000418884300024PubMed ID: 29306245Background/aim: The Motor Function Measure (MFM-32) is a classification system for ambulant and nonambulant patients with neuromuscular diseases (NMDs). We aimed to translate it into Turkish, culturally adapt it, and test its reliability and validity for Turkish patients with NMDs. Materials and methods: The translation of the 32 items assessing three functional areas: standing position and transfers (D1: 13), axial/proximal (D2: 12), and distal (D3: 7) motor functions was performed according to the established guidelines for cross-cultural adaptation. Totally 51 patients (12.56 +/- 8.84 years; F/M 12/39) were tested. Vignos and Brooke scores for the lower and upper extremities, respectively, were used for the validity of the MFM-32-TR items, which were rated on a 4-point Likert scale. Results: The agreement coefficients for interrater reliability were excellent (0.72-0.93) for 10 items, good (0.58-0.77) for 16 items, and moderate (0.42-0.56) for 6 items of the MFM-32-TR. The intertester reliability varied from good to excellent and the intraclass correlation coefficient was 0.76-0.93. The MFM-32-TR positively correlated with Vignos and Brooke scores with coefficients 0.47 to 0.75, indicating concurrent validity. Conclusion: The MFM-32-TR is a reliable and valid outcome measure for the assessment of motor function of people with NMDs in our sociocultural context

Turkish version of the Motor Function Measure Scale (MFM-32) forneuromuscular diseases: a cross-cultural adaptation, reliability, and validity study

WOS: 000418884300024PubMed ID: 29306245Background/aim: The Motor Function Measure (MFM-32) is a classification system for ambulant and nonambulant patients with neuromuscular diseases (NMDs). We aimed to translate it into Turkish, culturally adapt it, and test its reliability and validity for Turkish patients with NMDs. Materials and methods: The translation of the 32 items assessing three functional areas: standing position and transfers (D1: 13), axial/proximal (D2: 12), and distal (D3: 7) motor functions was performed according to the established guidelines for cross-cultural adaptation. Totally 51 patients (12.56 +/- 8.84 years; F/M 12/39) were tested. Vignos and Brooke scores for the lower and upper extremities, respectively, were used for the validity of the MFM-32-TR items, which were rated on a 4-point Likert scale. Results: The agreement coefficients for interrater reliability were excellent (0.72-0.93) for 10 items, good (0.58-0.77) for 16 items, and moderate (0.42-0.56) for 6 items of the MFM-32-TR. The intertester reliability varied from good to excellent and the intraclass correlation coefficient was 0.76-0.93. The MFM-32-TR positively correlated with Vignos and Brooke scores with coefficients 0.47 to 0.75, indicating concurrent validity. Conclusion: The MFM-32-TR is a reliable and valid outcome measure for the assessment of motor function of people with NMDs in our sociocultural context

TÜRK TOPLUMUNDA EN SIK RASTLANILAN MİYOFOSFORİLAZ (PYGM) MUTASYONLARI: MCARDLE HASTALIĞININ GENETİK TANISI İÇİN YENİ NESİL DİZİLEME

Amaç: Miyofosforilaz enzimini kodlayan PYGM genindeki mutasyonlar, enzimde aktivite eksikliğine neden olarak glikojen depo hastalığı tip V (GDHV – McArdle hastalığı) fenotipini ortaya çıkarmaktadır. Dünya genelinde en sık rastlanılan PYGM mutasyonları sırasıyla p.Arg50X ve p.Arg488X nokta mutasyonları olup, proteinde erken sonlanmaya neden olan anlamsız mutasyonlardır. Bu çalışmada ülkemizde genetik tanının konulabilmesi ve bu testlerin tanı akışında kas biyopsisinin önüne geçebilmesi için, Türk McArdle hastalarında PYGM geninde en sık görülen mutasyonların yeni nesil dizileme (YND) teknikleri ile belirlenmesi ve moleküler yöntemler ile bir tanı algoritması oluşturulması amaçlanmaktadır. Gereç ve Yöntem: Çalışma, daha önce kas biyopsisinde enzim histokimya ile miyofosforilaz enzim eksikliği tanısı konmuş 19 aileden oluşan hasta ve aile fertleri ile yürütülmüştür (n=34). Kontrol grubu sağlıklı hastalarla akrabalık ilişkisi bulunmayan ve miyopati klinik bulguları taşımayan kişilerden oluşturulmuştur (n=53). Hastalardan elde edilen DNA Illumina Miseq sisteminde YND ile dizilenmiştir. Dizileme verileri Burrows-Wheeler Aligner ile insan genom sekansına hizalanmış (hg19), BAM dosyaları SAMtool ile okunmuş, tek nükleotid polimorfizmler ve küçük insersiyon-delesyonların tespitinde GATK, fonksiyonel anotasyonların tespitinde SnpEff ve varyant filtreleme için VarSifter kullanılmıştır. Bulgular: Çalışma grubumuzda PYGM’de en çok rastlanılan mutasyon p.Met1Val/c.1A>G (hasta/aile=18/7) nokta mutasyonudur ve daha önce Türk kökenli bir hastada saptanmıştır. Çalışmada tespit edilen diğer mutasyonlar, p.621Phe_622Iledel/c.1864_1865delCTT (6/4), yeni keşfedilen g.4853_4854delTG (4/3), p.Arg488X/c.1462C>T (3/3) ve p.665Lys_666Glnfs/c.1998_1999delA (2/1) olarak sıralanmıştır. Bunların dışında, dünya genelinde hot-spot olan p.Arg50X (1) erken sonlanma mutasyonu sadece tek bir hastada belirlenmiştir. Sonuç: Türk GDHV hastalarında en sık rastlanılan p.Met1Val/c.1A>G mutasyonu, başlangıç kodonunu ortadan kaldırdığından, enzim translasyonunu yok ederek McArdle fenotipinin ortaya çıkmasına neden olmaktadır. Hastalarda, daha az olmakla birlikte, 5 ayrı mutasyon daha belirlenmiştir. Bu çalışmada g.4853_4854delTG mutasyonu 3 farklı ailede saptanan yeni bir mutasyon olup, diğerleri daha önce başka çalışmalarda tespit edilmiştir. YND teknikleri ile, Türkiye’den 19 ailede ortak PYGM mutasyonları belirlenmiş ve moleküler tanı için bir akış şeması oluşturulmuştur

SNUPN deficiency causes a recessive muscular dystrophy due to RNA mis-splicing and ECM dysregulation

SNURPORTIN-1, encoded by SNUPN, plays a central role in the nuclear import of spliceosomal small nuclear ribonucleoproteins. However, its physiological function remains unexplored. In this study, we investigate 18 children from 15 unrelated families who present with atypical muscular dystrophy and neurological defects. Nine hypomorphic SNUPN biallelic variants, predominantly clustered in the last coding exon, are ascertained to segregate with the disease. We demonstrate that mutant SPN1 failed to oligomerize leading to cytoplasmic aggregation in patients' primary fibroblasts and CRISPR/Cas9-mediated mutant cell lines. Additionally, mutant nuclei exhibit defective spliceosomal maturation and breakdown of Cajal bodies. Transcriptome analyses reveal splicing and mRNA expression dysregulation, particularly in sarcolemmal components, causing disruption of cytoskeletal organization in mutant cells and patient muscle tissues. Our findings establish SNUPN deficiency as the genetic etiology of a previously unrecognized subtype of muscular dystrophy and provide robust evidence of the role of SPN1 for muscle homeostasis