Type 2 innate lymphoid cells have a significant impact on chronic atopic dermatitis by regulating skin tissue fibrosis

이지호 울산대학교 일반대학원 2025 국내석사

Atopic dermatitis is a chronic dermatitis disease, 10% of children and 1-3% of adults are diagnosed. Although it develops due to various complex causes, it is known to be greatly influenced by Th1 and Th2 cells. Depending on the type of acute and chronic atopic dermatitis. The influence of Th1 and Th2 cells are different function and the type of cytokines act dominantly varies. In chronic atopic dermatitis to assisted Th1 cells and type 1 cytokines are usually known as their dominant mechanism. In acute atopic dermatitis, type 2 innate lymphoid cells(ILC2) are known to affect atopic dermatitis, and mechanisms suggest that ILC2 is stimulated by type 2 cytokines such as interleukin-25(IL-25) and Interleukin-33(IL-33), which are secreted by keratinocytes, producing interleukin-4 (IL-4) and interleukin-13 (IL-13). In addition, atopic dermatitis is obligatory accompanied by fibrosis of the dermis, so we would check whether ILC2 affects the fibrosis of the dermis. As far as it's known, fibroblasts have been increasing collagen production by Th2-derived cytokines, resulting in fibrosis. ILC2 is secrete which type 2 cytokines, is also thought to affect skin fibrosis. We used C57BL/6 mice to treat Aspergilus fumigatus for 5 days and progress to 2 days rest for 5 weeks to make chronic atopic dermatitis. and we used transepidermal water loss, skin clinical symptom score , gene expression, histological analysis, collagen assay, total serum IgE, flow cytometrys analysis. Then, to determine whether ILC2 affects atopic dermatitis, chronic atopic dermatitis has been verified using Rag1-/- mice, An experiment was conducted to verify that atopic dermatitis was alleviated in the absence of ILC2 by continuously injecting anti-CD90.2 into Intraperitoneal at intervals of 200µg every two days before treatment of the Af extract into the Rag1-/-mice. As a result, Rag1-/-mice induced chronic atopic dermaltitis models, similar result were found in transepidermal water loss, skin clinical symptom score , gene expression, histological analysis, collagen assay, total serum IgE, flow cytometrys analysis to like wild type C57BL/6 mice. key words : Chronic atopic dermartitis, helper T cells, type 2 cytokines, type 2 Innate lymphoid cells

Th1, Th2형 cytokine이 인간 비점막섬유아세포의 eotaxin, RANTES 생산과 mRNA 발현에 미치는 영향

송민 경희대학교 대학원 2002 국내박사

Background and Objectives; Fibroblast, a rich source of chemokines, interact with eosinophil and play a key role in the pathogenesis of airway diseases. There have been several reports about the effect of Th1, Th2 cytokines on eotaxin and RANTES production by human dermal and lung fibroblasts, but few studies have been reported about the effect of them by human nasal fibroblasts(HNF). The aims of study were to investigate whether Th1 or Th2 type cytokines are able to induce the expression and the production of eotaxin and RANTES in HNF and to verify the correlation between the stimulation of different cytokines and chemokines in HNF. Materials and method; Inferior turbinate mucosal samples were collected from 5 patients with nasal septal deviation at the time of surgery. Patients who had received systemic and topical steroids, antihistamines or antibiotics within 2 months before surgery and patients with allergy, chronic sinusitis, nasal polyps were excluded from this study. Cultured HNF were stimulated by IL-13, TNF-α, IFN-γ, IL-13+TNF-α, IL-13+IFN-γ, TNF-α+IFN-γ for 6, 24, 48 hours. They were compared with HNF cultured without any stimulation. In addition to it, HNF were stimulated by different concentration of IL-13 (0.2, 2, 20, 200 ng/ml) for 6 hours. Cells were collected and analyzed in terms of mRNA expression of eotaxin and RANTES respectively using RT-PCR. From the supernatant, we investigated the production of eotaxin and RANTES using ELISA. Results; TNF-α and IFN- γ(Th1 cytokines) induced the mRNA expression and the production of RANTES in HNF and they synergistically activated HNF to produce RANTES. The time-dependent increase of RANTES expression was revealed. IL-13(Th2 cytokines) induced the mRNA expression and the production of eotaxin in HNF and it produced it synergistically with TNF-α or IFN-γ. IL-13 induced the expression and the production of eotaxin in HNF with the increase of concentration. Conclusion; This study suggests that Th1 or Th2 type cytokines induce the expression and the production of eotaxin and RANTES in human nasal fibroblasts. TNF-α or IFN-γ induce more RANTES than eotaxin but IL-13 induces more eotaxin than RANTES. There may be some synergy of cytokines for the expression and production of chemokines. From the different expression timing of two chemokines, eotaxin attracts eosinophil earlier and RANTES attracts it later. Because two chemokines activate eosinophil through the CCR3 receptor, the development of an antagonist for this receptor may reduce tissue eosinophilia.

Hydroxysafflor yellow A는 천식 쥐 모델에서 nuclear factor-kappa B의 활성을 억제함으로써 알레르기성 기도 염증을 약화시킨다

설천비 전북대학교 일반대학원 2016 국내석사

Safflower is the dry flower of Carthamustinctorius L. and naturally distributed around the world. Hydroxysafflor yellow A (HSYA), the main active ingredient in safflower, is shown to reduce several inflammatory responses mediated by various stimuli or cytokines. Thus, the objective of this study is to determine whether the HSYA attenuates inflammatory response in an ovalbumin (OVA)-induced asthma model. OVA-sensitized /challenged mice represented the inflammatory cell infiltration in lung tissues as well as airway hyperresponsiveness (AHR) to in haled methacholine. In addition, these mice had an increased amount of eosinophils, T-helper2 type cytokines [interleukin(IL)-4, IL-5, and IL-13], eotaxin, and adhesion molecules in their bronchoalveolar lavage fluids. However, the administration of HSYA before the OVA challenge resulted in significant inhibition of these asthmatic reactions. Moreover, OVA significantly increased the transcriptional activity of nuclear factor-kappa B (NF-κB) in lung tissues, whereas HSYA markedly suppressed NF-κB translocation. These findings indicate that HSYA protects against OVA-induced airway inflammation and AHR, at least in part via downregulation of NF-κB activity. These data support the utility of HSYA as a potential medicine for the treatment of asthma. Keywords : Hydroxysafflor yellow A, Asthma, Airway inflammation, T-helper2 type cytokine, and NF-κB

Hookworms Dynamically Respond to Loss of Type 2 Immunity

Ferguson, Annabel Ansel University of Pennsylvania ProQuest Dissertations 2024 해외박사(DDOD)

소속기관이 구독 중이 아닌 경우 오후 4시부터 익일 오전 9시까지 원문보기가 가능합니다.

Hookworms are highly successful parasites, having coevolved with humans since we have been a species. This has culminated in a balance of hookworm traits that promote growth and fecundity with those that enable host tolerance of the infection. This project explores the coevolved traits from hookworms and the host immune response to worms. Type 2 immunity targets infections with parasitic worms, and is comprised of activities that directly harm or eject worms from their hosts. Many of these are dependent on the targets of the transcription factor Signal transduction and Activator of Transcription 6 (STAT6), which gets activated by signaling cytokines Interleukin (IL) -4 and -13. To counter this, worms possess a large repertoire of excreted and secreted (E/S) products that are released into the host environment and enable parasitism of the host, by both allowing tissue migration and by suppressing the host immune response. This work sought to test the central hypothesis that STAT6-dependent immune responses have selected hookworm traits that enable host tolerance of this infection. It did so by experimentally evolving hookworms in STAT6 KO hosts via serial passaging, and then analyzing worm traits as well as the host immune response to infection with adapted worms. The hookworms derived from STAT6 KO hosts were larger and more fecund, but had a contracted array of E/S proteins. This suggested that size and reproduction were enhanced over the expression of E/S when hosts lack an immune response. Surprisingly, this adaptation did not result in loss of hookworm traits that promoted growth, survival and reproduction when STAT6 immune responses were restored via the infection of WT mice with STAT6 KO-adapted worms. However, during this infection, the hosts displayed an increased inflammatory immune response in both the lungs and the gut, which likely led to increased mortality of the hosts. This work suggests that a persistent evolutionary pressure of STAT6 dependent immune responses is needed for maintaining immune-suppressing traits in hookworm populations.

Prostate Fibrosis: A Consequence of Prostate Inflammation and a Potential Etiology of Urinary Dysfunction

Bell-Cohn, Ashlee Northwestern University ProQuest Dissertations & T 2020 해외박사(DDOD)

소속기관이 구독 중이 아닌 경우 오후 4시부터 익일 오전 9시까지 원문보기가 가능합니다.

(The) modulation of host immune responses by porcine circovirus type 2 ORF2 and ORF3

최창용 Graduate School, Korea University 2017 국내박사

Porcine circovirus type 2 (PCV2) is the main etiological agent of postweaning multisystemic wasting syndrome (PMWS). The mechanism of pathogenicity associated with PCV2 infection is still not fully understood. Nevertheless, the fact that large amounts of proinflammatory cytokines within lymphoid tissues are released during the early stage of PCV2 infection may induce chronic inflammatory responses followed by the destruction of lymphoid tissues. However, how PCV2 infection causes an excessive inflammatory response in the host immune system during the early stage of PCV2 infection is still not elucidated. Defining how each ORF of PCV2 manipulates the host immune system may be helpful to understand disease progression of PMWS. In this study, it was found that PCV2 ORF3 directly interact with RGS16 in the cytoplasm of host epithelial cells and its interaction leads to ubiquitin-mediated proteasomal degradation of RGS16. Facilitated degradation of the RGS16 by PCV2 ORF3 further enhances NFB translocation into the nucleus through the ERK 1/2 signaling pathway and increased secretion of IL-6 and IL-8 proinflammatory cytokine. Consequently, more severe inflammatory responses and leukocyte infiltration occur around PCV2 infection site. Furthermore, direct interaction between the PCV2 ORF2 and C1QBP within the cytoplasm of host macrophages was demonstrated in this study. The physical interaction between PCV2 ORF2 and C1QBP inhibits ubiquitin-mediated proteasomal degradation of C1QBP in macrophages. Increased stability of the C1QBP by the interaction with PCV2 ORF2 further enhances phagocytic activity of porcine macrophages through the phosphoinositol-3-kinase (PI3K) signaling pathway. These results may provide the first evidence showing how PCV2 infection facilitates the expression of proinflammatory cytokines, enhances phagocytic activity of macrophages and creates a more severe inflammatory response during the early stages of PCV2 infection.

Anti-allergic and anti-inflammatory activity of the aqueous extract of Diospyros kaki calyx

박해란 우석대학교 일반대학원 2017 국내박사

감꼭지 물 추출물의 항알레르기 및 항염증작용 박해란 우석대학교 대학원 약학과 (지도교수 : 신태용) 알레르기는 복잡한 병리적 현상의 결과로 면역반응에 따른 인체의 전신적 또는 국소적인 장애이다. 제Ⅰ형 알레르기반응의 기전은 인체에 유입된 외인성 항원 또는 생체에서 유래한 내인성 항원이 IgE 항체를 생성하고, 생성된 IgE항체는 호염기구나 비만세포의 세포막 표면에 있는 IgE 수용체에 결합되어 감작이 성립된다. 동일한 항원이 체내에 다시 유입되면 호염기구나 비만세포에 결합되어 있는 IgE항체 사이에 가교가 형성되어 탈과립이 일어나 화학적 매개물질의 유리된다. 유리되는 중요한 화학적 매개물질은 이미 합성되어 저장되어 있다가 유리되는 히스타민, 헤파린 등과 비만세포막에 유래하는 아라키돈산이 대사되어 생성되는 SRS-A, 프로스타그란딘류 등이다. 유리되는 화학적 매개물질에 의해 평활근이 수축되고 모세혈관의 투과성이 항진되며 분비항진 등에 의해 기관지천식, 담마진, 알레르기성 비염 등이 일어나며 심하면 아나필락시 등의 조직 상해가 일어날 수 있다. 알레르기 치료약물로는 탈과립저해제와 화학전달물질 작용억제제 및 화학전달물질 합성 저해제 등이 임상에서 사용되고 있지만 장기 투약에 의해 여러 가지 부작용을 초래할 수 있다. 이에 저자는 부작용이 적으며 알레르기 치료 작용이 우수한 약물 개발을 목적으로 실험에 착수하였다. 실험에 사용한 감꼭지는 한방에서 딸꾹질과 구역질 등에 사용되고 있지만 항알레르기와 항염증작용에 대한 연구는 전무한 상태이다. 저자는 생체 내 및 생체 외 모델을 이용한 실험에서 감꼭지 물 추출물의 알레르기성 염증 치료효과를 확인 하였으며 그 기전을 규명 하였다. 그 결과는 다음과 같다. 1. AEDKC의 경구 투여는 전신성 아나필락시스 모델에서 마우스의 직장 체온의 감소를 농도 의존적으로 억제하였으며 OVA에 의해 증가된 혈청 히스타민, 총 IgE, OVA-특이적 IgE 및 IL-4를 감소시켰다. 2. PCA 모델에서 AEDKC는 Evans blue pigmentation를 감소시켰다. 3. AEDKC는 RBL-2H3 세포와 초대 배양 비만세포에서 세포내 칼슘의 감소를 통한 기전으로 히스타민과 β-hexosaminidase의 유리를 억제하였다. 4. AEDKC는 NF-κB의 감소 기전을 통해 염증성 사이토카인인TNF-α와 IL-4의 분비와 발현을 억제하였다. 이상의 결과 AEDKC는 히스타민의 유리를 억제하며 NF-κB의 활성화 억제와 세포내 칼슘 이온 농도의 억제로 TNF-α와 IL-4 등의 염증성 사이토카인의 발현과 분비를 억제하는 작용기전으로 항알레르기와 항염증작용을 나타낸다고 사료된다. Anti-allergic and anti-inflammatory activity of the aqueous extract of Diospyros kaki calyx Park, Hae Ran Dept. of Pharmacy Graduate School of Woosuk University Diospyros kaki calyx has been used for the treatment of hiccup and nausea in traditional Oriental medicine. The purpose of this study is to elucidate the inhibitory effect of aqueous extract of Diospyros kaki calyx (AEDKC) on mast cell-mediated immediate -type hypersensitivity and to underlying mechanism of action. For in vivo, ovalbumin (OVA)-induced active systemic anaphylaxis (ASA) and immunoglobulin E (IgE)-mediated passive cutaneous anaphylaxis (PCA) were used. The control drug dexamethasone was used to compare the efficacy of AEDKC. For in vitro, IgE-stimulated RBL-2H3 cells and primary cultured peritoneal mast cells were used to determine the role of AEDKC on the activation of mast cells. Oral administration of AEDKC dose-dependently suppressed rectal temperature decrease and increases in serum histamine, total IgE, OVA-specific IgE, and interleukin (IL)-4 in the active systemic anaphylaxis. In the PCA model, AEDKC reduced Evans blue pigmentation. AEDKC suppressed the release of histamine and β-hexosaminidase through the reduction of intracellular calcium in RBL-2H3 cells and primary cultured mast cells. In addition, AEDKC decreased the expression and secretion of TNF-α and IL-4 by the reduction of nuclear factor (NF)-κB. These results provide evidence that AEDKC contributes to the prevention or treatment of mast cell-mediated immediated-type hypersensitivity. The author suggest that AEDKC may be a potential candidate for the treatment of mast cell-mediated allergic diseases.

Effects of the antidiabetic DPP4/CD26 inhibitors evogliptin and sitagliptin on immune function of H9 T helper cells

윤현이 Greduate School, Korea University 2021 국내박사

Dipeptidyl peptidase (DPP) 4/CD26 is a multifunctional protein containing an ecto-domain with DPP4 enzymatic activity and known to involve in T cell immune response, including T helper 1 cells. Numerous membrane DPP4 (mDPP4) is on the surface of T cells and soluble DPP4 (sDPP4), originating from mDPP4 exists at a high level in blood. Altered DPP4/CD26 expression in DPP4-riched T cell is associated with diverse autoimmune diseases including type 1 diabetes, rheumatoid arthritis (RA), multiple sclerosis (MS), and inflammatory bowel diseases (IBD). DPP4 inhibitors is the most prescribed antidiabetic drugs that suppress the enzymatic activity by direct binding at active site of DPP4. However, their adverse effects are still unclear on the immune function of DPP4-riched T helper cells under the treatment of patients with type 2 diabetes. The aim of this study was to assess whether antidiabetic drugs and other DPP4 inhibitors affect the dipeptidyl enzymatic activity of mDPP4 and intrinsic function of CD4+CD26+ H9 cells in terms of cytokine expression and cellular profiles. The mDPP4 enzymatic activity, T helper cell-specific cytokine expression, and cellular profiles, including cell counts, cell viability, DNA synthesis, and apoptosis, were estimated in pokeweed mitogen (PWM)-activated CD4+CD26+ H9 cells with or without the antidiabetic drugs evogliptin and sitagliptin or DPP4 inhibitors such as diprotin A and berberine. PWM treatment alone stimulated strongly the expression of T helper cell-specific cytokines such as interleukin (IL)-2, IL-10, IL-13, granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-gamma (IFN-ɣ), and tumor necrosis factor-alpha (TNF-α) in CD4+CD26+ H9 cells, albeit induced marginally the expression of CD26. The treated evogliptin or sitagliptin potently inhibited mDPP4 enzymatic activity but did not influence either T helper cell-specific cytokine expression or cellular profiles in PWM-activated CD4+CD26+ H9 cells. In contrast, diprotin A inhibited transiently mDPP4 enzymatic activity and berberine had a little or no inhibitory ability with cytotoxicity to cells. These results imply that mDPP4 enzymatic activity is not essential for cell signaling pathways for T helper cell-specific cytokine expression upon immune stimulation. Moreover, the results implicates that immune functions such as T helper cell-specific cytokine expression are performed normally even after the treatment with evogliptin or sitagliptin which inhibits strongly mDPP4 enzymatic activity.

Different composition of serum amino acids and bile acids in patients with impaired fasting glucose and type 2 diabetes mellitus compared with normal controls

이상국 Graduate School, Yonsei University 2017 국내박사

배경 : 최근 메타볼로믹스 분석기법의 발달로 새로운 바이오마커의 개발에 지속적인 진보가 있어왔다. 새로운 바이오마커들 중 혈중 아미노산의 구성은 여러 질환들에서 유망한 바이오마커로써의 가능성을 보여주었다. 특히 분지사슬 아미노산(branched chain amino acid)의 증가가 제2형 당뇨병, 비만, 심장혈관질환에서 일관되게 보고되고 있다. 뿐만 아니라, 최근의 연구들에서는 담즙산이 포도당 대사에서 중요한 역할을 한다는 사실이 보고되었다. 따라서 공복혈당장애나 제2형 당뇨병을 가진 환자들에서의 특징적인 공복 혈중 아미노산과 담즙산 구성의 차이를 찾아내기 위해 건강한 대조군과 공복혈당장애 또는 제2형 당뇨병을 가진 환자들에서 혈중 아미노산과 담즙산의 구성을 분석하고자 하였다. 방법 : 당뇨약제를 복용하지 않은 제2형 당뇨병으로 최초 진단된 72명, 공복혈당장애 환자 97명, 정상인 75명에서 총 20종의 아미노산과 15종의 담즙산의 농도를 고성능액체크로마토그래피-질량분석기 법으로 측정하였다. 혈청 TNF-α와 IL-6의 농도는 상품화된 ELISA 키트를 이용하여 측정하였다. C2C12 마우스 근원세포를 사용하여 팔미테이트 처리를 통해 인슐린 저항성을 유발하고 MAFbx와 MuRF1의 발현의 변화를 평가하였다. 결과 : 정상인에 비해 공복혈당장애와 제2형 당뇨병 환자에서 공복 혈청 분지사슬 아미노산뿐만 아니라 글루탐산, 리신, 페닐알라닌, 아르기닌, 알라닌, 티로신, 아스파르트산이 함께 증가하는 것을 발견하였다. 이러한 아미노산의 증가는 공복혈당장애보다 제2형 당뇨병 환자에서 더 크게 나타났다. 증가된 아미노산들의 농도는 공복혈당, HOMA-IR, 염증성 사이토카인과 우수한 양의 상관성을 보였다. 그리고 HOMA-IR과 염증성 사이토카인은 공복 혈중 아미노산 농도를 결정하는 중요한 두 가지 독립적인 요소였다. C2C12 세포를 이용한 체외실험결과, 팔미테이트의 처리는 인슐린 저항성과 염증성 사이토카인 유전자의 발현을 촉진하였고 MAFbx 유전자 및 단백질의 발현 증가를 확인하였다. 마지막으로 글리신 포합 담즙산의 경우 정상인에 비해 공복혈당장내나 제2형 당뇨병 환자에서 감소함을 확인하였다. 결론 : 공복 혈중 아미노산의 상승은 인슐린 저항성의 원인요소이기 보다 인슐린 저항성의 조기증후일 가능성이 높다. 이러한 혈중 아미노산 상승의 기전으로는 골격근에서의 근육 단백질의 가수분해 증가가 유력하다. 당뇨병 환자에서의 글리신 포합 담즙산의 감소된 결과를 통해 장에서의 담즙산 포합조절이 포도당 대사에서 중요한 역할을 수행할 가능성을 확인하였다. Background: Recent advance of metabolomics has made remarkable progress in discovery of novel biomarkers. Among these biomarkers, blood amino acids (AAs) profiles have produced a number of promising results. Elevated blood levels of AAs, especially branched chain amino acids (BCAAs) have been reported in Type 2 DM (T2DM), obesity, and cardiovascular disease. In addition, emerging researches shows the importance of bile acids (BAs) in glucose homoeostasis. Therefore, we performed AAs and BAs profiling in patients with impaired fasting glucose (IFG) and T2DM as well as healthy control to find specific alteration of these profile in patients with IFG and T2DM. Methods: Twenty AAs and fifteen kinds of BAs were analyzed in samples from 72 T2DM patients without diabetic medications, 97 patients with IFG and 75 healthy control subjects using high performance liquid chromatography (HPLC) - tandem mass spectrometry. Serum TNF-α and IL-6 were measured using a commercially available human ELISA kits. The C2C12 mouse myoblast cell lines were used to examine the changes of MAFbx and MuRF1 expressions after development of insulin resistance induced by palmitate treatment. Results: Fasting serum AAs, not only BCAAs but also glutamic acid, lysine, phenylalanine, arginine, alanine, tyrosine, aspartic acid are higher in patients with T2DM and intermediate in patients with IFG compared with normoglycemic controls. These serum AAs concentrations positively correlated with fasting glucose, HOMA-IR, and pro-inflammatory cytokines (TNF-α and IL-6). In addition, HOMA-IR and pro-inflammatory cytokines were two important independent predictor of blood AAs level. In vitro experiments showed that palmitate treatment in C2C12 myotubes induced insulin resistance, increased pro-inflammatory cytokine gene expression, and increased MAFbx gene and protein expression. Finally, we found that glycine conjugated BAs were decreased in the patients with IFG and T2DM. Conclusion: Fasting blood AAs increase can be early manifestation of insulin resistance. Increased muscle proteolysis by insulin resistance and inflammatory cytokines is possible mechanism of these AAs elevation. Glycine conjugated BAs are decreased in the patients with IFG and T2DM suggesting control of glycine conjugation and deconjugation of BAs could affect glucose homeostasis.

New biological functions of apoptozole and bioimaging using fluorescent probes

백경화 Graduate School, Yonsei University 2014 국내박사