아토피피부염의 피부거주 T 세포에서 CCCTC-결합 인자의 역할

의과학Atopic dermatitis (AD) is a highly pruritic, chronic relapsing inflammatory skin disease characterized by significant T-cell infiltration and frequently recurs in the same areas of the skin, and even resolved skin keeps the disease’s memory. While the role of tissue-resident memory T cells (TRM cells) has been well studied in the chronic inflammatory skin diseases such as psoriasis which relapses in the same sites, that of TRM cells in AD has not been investigated in detail. CCCTC-binding factor (CTCF) is a ubiquitously expressed regulator of fundamental genetic processes including transcription, intra- and interchromosomal interactions, and chromatin structure. Because of its critical role in genome function, CTCF binding patterns have long been assumed to be largely invariant across different cellular environments. Therefore, the functional analysis of CTCF in the different types of cells and tissues would further expand our knowledge in the roles of CTCF. In this dissertation, I focus on the role of CTCF in TRM cells in AD. To explore the function of TRM cells in AD. I examined how many TRM cells expressing the canonical maker CD69 and CD103 existed and which cytokines including IL-4, IL13, IL-17, IL-22 and IFN-ƴ these cells produced in human AD and age-matched normal skin tissues and in AD-like mouse models generated with the treatment of allergens (1-Fluoro-2,4-dinitrobenzene (DNFB), oxazolone, ovalbumin) or mixture thereof. I also evaluated which AD triggering factors affected the development of TRM cells and the production of cytokines using purified cutaneous lymphocyte-associated antigen (CLA) expressing T cells from normal peripheral blood mononuclear cells (PBMCs). Finally, using transcriptomics, I evaluated the cytokine signatures and novel genes associated with skin TRM cells compared to migratory memory T cells (TMM) in human normal and AD skin. I observed that CD69+ and CD103+ TRM cells significantly infiltrated into chronic AD skin compared to normal skin. However, normal skin mostly ontained CD103- and CD69- T cells. Significant numbers of CD69+ AD TRM cells produced Th2 cytokines (IL-4, IL-13), Th17 cytokines (IL-17, IL-22), and Th1 cytokines (IFN-ƴ). In AD-like mouse model induced by allergen mixture, CD69+ skin TRM cells produced various cytokines (IL-4, IL-17, IL-22 and IFN-ƴ) compared to a single allergen induced AD-like mouse models. During the incubation of CLA+ T cells for 3 weeks, CD69+ expression of CD4+ and CD8+ T cells was continuously increased in a time-dependent manner while CD103+ expression was not significantly changed. Repeated exposure to thymic stromal lymphopoietin (TSLP), a well-known AD triggering factor, induced more CD69+ TRM cells during the development of AD TRM. Various cytokines (IL4, IL17, IL-22 and IFN-ƴ) were considerably produced in CD69+ TRM cells compared to CD69- T cells in AD mimicking mice after 3 weeks. I further confirmed that AD CD69+ TRM cells expressed higher levels of Th1, Th2, Th17 and Th22 cytokines and their transcription factors (T-bet, GATA3, RORƴt and Ahr) showing unique transcriptional profiles distinct from those of AD CD69- TMM cells or normal skin CD69+ TRM cells. I further identified various genes associated with tissue egress and residency specific to AD TRM cells. Among those, I chose CCCTC-binding factor (CTCF) because of its role in immune diseases. I confirmed the decrease of CTCF in CD69+ TRM cells from AD patient skin. In conclusion, CTCF may play an important role in regulating the expression of multiple cytokines from CD69+ TRM cells in AD skin. Through this mechanism, repeated exposure to AD-triggering factors might induce AD TRM cells to produce multiple cytokines and sustain the recurrence and chronicity of AD patients. 아토피피부염은 소양증을 동반한 만성 재발성의 염증성 질환으로, 외부환경으로부터 침입한 항원 물질과 진피의 면역세포에 의해 야기되는 면역반응과 피부의 같은 자리에 계속해서 다시 발생하는 것을 특징으로 한다. 피부 거주 T 세포 (TRM 세포)는 기억 T 세포의 한 종류로 말초 조직에 오랜 기간 동안 계속해서 존재하는 세포로 알려져 있으며, 위장관, 기도, 피부와 같은 숙주와 환경 사이의 접점의 상피 막 조직에 존재한다. CCCTC-binding factor (CTCF)는 DNA 결합 zinc-finger 단백질로서, 고차원 크로마틴 구조 결정에 중요하게 관여하여 다양한 분자생물학적 기전을 통해 유전자 발현을 조절한다고 알려져 있는 유전자이다. 본인은 아토피피부염에서의 피부 거주 T 세포의 역할과 성상을 확인하고자 하였고, 피부 거주 T 세포가 아토피피부염을 악화시키는데 있어서 CTCF 단백질의 발현 및 기능을 연구하였다. CD69+ 및 CD103+ TRM 세포가 정상 피부조직에 비해 아토피피부염 환자의 피부조직에 현저히 침투하는 것을 관찰하였고, 정상 피부조직에는 비교적 CD103- 및 CD69- 세포가 많이 발현되는 것을 확인하였다. 또한, 면역형광염색법을 통해 CD69+ AD TRM 세포가 Th2 사이토카인 (IL-4, IL-13), Th17 사이토카인 (IL-17, IL-22) 및 Th1 사이토카인 (IFN-ƴ)과 같은 다양한 사이토카인들을 분비하는 것을 확인하였다. 아토피피부염과 유사한 마우스 모델 (DNFB, 옥사졸론, 오발부민) 피부조직의 CD69+ TRM 세포에 비해 다양한 알러젠에 노출되는 아토피피부염 환자와 유사하게 이 알러젠들을 모두 혼합하여 유발한 마우스 모델 피부조직의 CD69+ TRM 세포에서 역시 다양한 사이토카인들을 많이 분비하는 것을 확인하였다. 3주간의 CLA+ T 세포 배양 동안, CD4+ 및 CD8+ T 세포의 CD69+ TRM 세포의 발현은 시간이 지날수록 지속적으로 증가 하였고, 아토피피부염과 유사한 환경을 조성해 주었을 때, 더 많은 CD69+ TRM 세포 및 다양한 사이토카인들이 생성되는 것을 확인하였다. 이를 증명하기 위해 AD CD69- TMM 및 CD69+ TRM 세포에서 RNA를 얻어 microarray 분석을 진행하였다. 그 결과 CD69+ TRM 세포에서 Th1, Th2, Th17 및 Th22 사이토카인과 그 전사인자 (T-bet, GATA3, RORƴt 및 Ahr)들이 높은 수준을 나타내는 것을 확인하였다. 전체 47322 개의 유전자 중 AD CD69- TMM 세포와 비교하였을 때 AD CD69+ TRM 세포에서 CTCF 유전자가 뚜렷하게 감소되었다. 이를 검증하기 위해, 본인은 CTCF 단백질의 발현이 정상인의 피부조직에 비해 아토피피부염 환자의 피부조직에서, 또한 CD69- TMM 세포에 비해 CD69+ TRM 세포에서 현저하게 감소됨을 면역화학염색법과 면역형광염색, real-time PCR 및 shRNA를 통해 확인하였다. 결론적으로, 본 연구를 통해 아토피피부염 유발 원인에 반복적으로 노출되면 CTCF 유전자 발현의 감소되어 아토피피부염의 CD69+ TRM 세포를 조절하게 됨을 확인하였고, 이로 인해 CD69+ TRM 세포가 다양한 사이토카인들을 생성하게 되어서 결론적으로 아토피피부염의 악화를 유도하게 됨을 확인하였다.open박

Positive reactions to nickel on a patch test do not predict clinical outcome of nickel alloy-based atrial septal defect occluder implantation

BACKGROUND: Patch testing is thought to be necessary prior to metal device implantation to rule out metal allergy-related complications; however, there are controversies over the effects of nickel allergy on the outcome of nickel alloy-based device implantation. OBJECTIVE: This study aimed to evaluate the adverse events in a Korean population of nickel allergy patients who underwent atrial septal defect (ASD) closure with a nickel-titanium alloy-based device. METHODS: We retrospectively reviewed the medical records of patients who underwent ASD closure with a nitinol device. RESULTS: Among 38 patients who had ASD closure, 4 of 5 nickel-allergic patients and 10 of the 33 non-nickel-allergic patients had post-closure complications. All patients fared well, without device failure culminating in device removal. CONCLUSION: In this study, positive reactions to nickel in a patch test were not associated with adverse early or late outcomes following ASD closure with a nickel alloy-based device.ope