Standard immunosuppressive treatment reduces regulatory B cells in children with autoimmune liver disease

IntroductionAutoimmune hepatitis (AIH) is a chronic liver disease caused by a perturbed immune system. The scarcity of short- and long-term immune monitoring of AIH hampered us to comprehend the interaction between immunosuppressive medication and immune homeostasis.Methods and patientsWe recruited children with AIH at the time of diagnosis and at the 1st, 3rd, 6th, 12th, 18th, and 24th months of immunosuppression (IS). We also enrolled children with AIH being on IS for >2 years. Children with drug-induced liver injury (DILI), and those receiving tacrolimus after liver transplantation (LT), were enrolled as disease/IS control subjects. Healthy children (HC) were also recruited. Peripheral blood mononuclear cells (PBMCs) were isolated from all participants. Healthy liver tissue from adult donors and from livers without inflammation were obtained from children with hepatoblastoma. By using flow cytometry, we performed multi-parametric immune profiling of PBMCs and intrahepatic lymphocytes. Additionally, after IS with prednisolone, tacrolimus, rapamycin, or 6-mercaptopurine, we carried out an in vitro cytokine stimulation assay. Finally, a Lifecodes SSO typing kit was used to type HLA-DRB1 and Luminex was used to analyze the results.ResultsUntreated AIH patients had lower total CD8 T-cell frequencies than HC, but these cells were more naïve. While the percentage of naïve regulatory T cells (Tregs) (CD4+FOXP3lowCD45RA+) and regulatory B cells (Bregs, CD20+CD24+CD38+) was similar, AIH patients had fewer activated Tregs (CD4+FOXP3highCD45RA-) compared to HC. Mucosal-associated-invariant-T-cells (MAIT) were also lower in these patients. Following the initiation of IS, the immune profiles demonstrated fluctuations. Bregs frequency decreased substantially at 1 month and did not recover anymore. Additionally, the frequency of intrahepatic Bregs in treated AIH patients was lower, compared to control livers, DILI, and LT patients. Following in vitro IS drugs incubation, only the frequency of IL-10-producing total B-cells increased with tacrolimus and 6MP. Lastly, 70% of AIH patients possessed HLA-DR11, whereas HLA-DR03/DR07/DR13 was present in only some patients.ConclusionHLA-DR11 was prominent in our AIH cohort. Activated Tregs and MAIT cell frequencies were lower before IS. Importantly, we discovered a previously unrecognized and long-lasting Bregs scarcity in AIH patients after IS. Tacrolimus and 6MP increased IL-10+ B-cells in vitro

Spondiloartropatili Hastalarda HLA Sınıf I Allel Sıklığı

Spondiloartropati (SpA), spinal inflamasyon ve periferal artrit ile daha az oranda da eklemdışı tutulumla karakterize, patogenezi henüz tam olarak bilinmeyen multisistemik bir gruphastalıktır. Genetik faktörlerin hastalığın gelişiminde önemli rolü vardır. 1973 yılındaBrevverton ve Schlosstein, HLA-B27 ile hastalık ilişkisini ortaya çıkarmıştır. HLA-B27 ilebirlikte diğer HLA moleküllerinin (DR1, DR4, DR8, DR15, A24, B39 ve B60)hastalıklailişkisi yapılan çalışmalarda gösterilmiştir. Bu bilgilerden yola çıkarak ve etnik farklılığında hastalıkla ilişkili olduğunu düşünerek SpA’lı Türk hastalarda HLA-A ve –B allellerininrolünü araştırmayı amaçladık. Hastalardan (n=784) heparinli kan örneği alındı. Lenfositizolasyonunu takiben komplemana bağımlı lenfositotoksisite yöntemi ile ticari kitler (Biotest-HLA-ABC tipleme plağı, 144X2, USA) kullanılarak tipleme yapıldı. Kontrol grubunun(n:1060) HLA-A ve –B doku grupları, serolojik veya moleküler yöntemler kullanılaraktespit edildi. Hastalarda HLA-B27 sıklığı %27 olarak saptandı. B27 negatif hastalar ile B27negatif kontroller karşılaştırıldığında, hastalarda HLA-A29 anlamlı olarak yüksek bulundu(p:0.0003, pc:0.004, OR:2.6, CI:1.5-4.4). HLA-B60 hastalarda anlamlı olarak yüksek (p:0.02,OR:0.5, CI:0.2-0.9) bulunmasına rağmen Bonferroni doğrulama testi sonrası istatistikselanlamlılık elde edilemedi (pc>0.05). B27 pozitif hastalarla kontrolleri karşılaştırdığımızdaHLA-A3 (p:0.0005, pc:0.008, OR:0.4, CI:0.3-0.7), HLA-B35 (p<0.0001, pc<0.003, OR:0.3,CI:0.2-0.4), HLA-B51 (P<0.0001, pc<0.003, OR:0.3, CI:0.2-0.6) ve HLA-B52 (p:0.001,pc:0.03, OR:0.04, CI:0.002-0.7) kontrol grubunda, HLA-B27 (p<0.0001, pc<0.003, OR:52,CI:36.2-74.7) ise hasta grubunda anlamlı olarak yüksek bulundu. Patogenezi tam olarakbilinmeyen SpA’ların gelişiminde genetik faktörlerin rolü büyüktür. HLA-B27 allelininhastalığın gelişimi ile ilişkili olduğu bilinmektedir. Etnik farklılıklarla birlikte diğer HLAmoleküllerinin de hastalığa karşı yatkınlık ve koruyuculukta etkili olabileceği yapılançalışmalarla gösterilmiştir