1,721,242 research outputs found
Translationally Controlled Tumor Protein in Chronic Spontaneous Urticaria
No full text학위논문(석사)--아주대학교 일반대학원 :의학과,2019. 2I. INTRODUCTION 1
II. MATERIALS AND METHODS 4
A. Clinical characteristics of the study subjects 4
B. Evaluation of clinical parameters of CSU patients 5
C. Measurement of TCTP in sera of study subjects 6
D. Purification of recombinant TCTP protein 7
E. Western blotting for detecting the dimerized TCTP in sera from CSU patients and NCs 8
F. Basophil activation test 8
G. Measurement of ß-hexosaminidase release from LAD-2 cells 9
H. Statistical analysis 10
III. RESULTS 12
A. Clinical characteristics of study subjects 12
B. Serum TCTP levels in CSU patients and NCs 12
C. Correlation of serum TCTP levels with clinical characteristics of chronic spontaneous urticaria 14
D. Effects of TCTP on basophil activation 21
E. β-hexosaminidase release from LAD2 cells upon the stimulation with TCTP 22
IV. DISCUSSION 24
V. CONCLUSION 28
REFERENCES 29MasterBackground and objectives: Translationally controlled tumor protein (TCTP) is widely expressed as housekeeping protein that has various intracellular and extracellular functions including its ability to activate mast cells. Extracellular TCTP has been found in nasal, bronchoalveolar lavage and skin blister fluids and exhibits the capacity to induce histamine release from basophils, suggesting that TCTP plays a role in allergic diseases including asthma, atopic dermatitis and urticaria. Dimerization of TCTP is important for its cytokine-like activity, therefore, we hypothesized that the dimerized TCTP may have an important role by inducing histamine and other mediator release, which may drive development of chronic spontaneous urticaria (CSU) symptoms.
Materials and Methods: One hundred sixteen CSU patients and 70 healthy normal controls (NCs) were enrolled at Ajou University Medical Center, Suwon, South Korea. Serum TCTP levels of study subjects were measured by ELISA and compared according to other CSU-related clinical parameters. Basophil activation test (BAT) was performed by measuring CD203c expression on peripheral basophils from CSU patients. In vitro, β-hexosaminidase release from human mast cell line, monomeric and dimeric TCTP stimulated LAD-2 mast cells was evaluated. Western blot (WB) analysis was performed to detect dimeric TCTP from sera of CSU patients and NCs in non-reducing condition. Rapid dot-blot immunoassay was used for measuring the level of IgG to high affinity Fc epsilon receptor I alpha subunit (FcεRIα) level in serum of CSU patients and NCs.
Results: There was no difference between serum levels of TCTP in CSU patients and NCs. The intensity of dimeric TCTP on WB was visually stronger in CSU patients than in NCs. Severe CSU patients had higher serum TCTP levels (P=0.049) and those having IgG positivity to FcɛRIα (P=0.038). Serum TCTP was significantly positive correlated with eosinophil cationic protein levels (Spearman’s rho 0.341, P=0.001). Both basophil and mast cell degranulation were significantly increased after the stimulation with dimeric TCTP, but, not with monomic TCTP.
Conclusion: The capability of TCTP to activate basophils and mast cells is dependent on its dimerization process, suggesting that the inhibition of TCTP dimerization might have therapeutic benefit for CSU. Association of TCTP levels with the presence of IgG to FcεRIα autoantibody indicating that autoimmune mechanisms of CSU which may related to its dimerization
Genetic Polymorphism of Prostanoid Receptors, CRTH2 and TBXA2R in Aspirin Hypersensitivity
No full text학위논문(박사)--아주대학교 일반대학원 :의학과,2010. 2-국문요약-
아스피린 천식 과민성 질환에서의 프로스타노이드 수용체인 CRTH2와 TBXA2R의 유전자 다형성 연구
아주대학교 대학원 의학과
나미 슈레스터 팔리케
(지도교수:박 해 심)
연구배경 및 목적: 아스피린 복용은 일부 사람들의 경우 아스피린 과민성 천식, 아스피린 과민성 두드러기, 만성 비염, 아나필락시스 같은 다양한 알레르기 반응을 유발하기도 한다. 본 연구는 아스피린 과민성으로 나타나는 대표적인 두 질환인 과민성 천식과 아스피린 과민성 두드러기에서 프로스타글란딘 D2 수용체, CRTH2,와 프로스타노이드 수용체, TBXA2R 유전자들에 대한 유전체 연구이다. 아스피린 과민성 천식 환자에서 호산구 염증 및 유입이 아스피린 내성 천식 환자에 비해 두드러지게 나타남을 고려해 볼때, 프로스타글란딘 D2 수용체인 CRTH2 유전자의 유전자 다형성에 따른 기능적 변화를 통한 호산구의 활성화가 아스피린 과민성 천식의 병인기전에 중요한 역할을 담당할 것으로 가정하고 CRTH2 유전자의 유전자 다형성 조사 및 아스피린 과민성 천식과의 연관성 연구를 수행하였다. 더욱이, 천식 및 아토피와의 연관성이 보고된 TBXA2R 유전자의 경우, 그 유전자의 유전자 다형성에 따른 기능 변화가 아스피린 과민성 두드러기의 병인기전과 관련되는지를 유전자 연관성 연구로 조사하였다.
재료 및 방법: 본 연구는 아주대 병원에 내원한 107명의 아스피린 과민성 천식 환자, 115명의 아스피린 내성 천식 환자, 그리고 133명의 정상대조군을 대상으로 CRTH2 유전자의 유전자 다형성 조사 및 연관성 연구를 수행하였다. TBXA2R 유전자의 경우, 아주대 병원에 내원한 167명의 아스피린 과민성 급성 두드러기 환자, 316명의 아스피린 과민성 만성 두드러기 환자, 그리고 265명의 정상대조군을 대상으로 수행하였다. 두 유전자의 유전형은 SNAPshot ddNTP primer extention 키트를 사용하여 결정되었다. 두 유전자의 유전자 다형성에 따른 기능 변환 연구는 luciferase reporter assay와 electrophoretic mobility shift assay로 수행하였다.
결과: 아스피린 과민성 천식 환자군에서 아스피린 내성 천식 환자군에 비해 높은 eotaxin-2 생성이 관찰되었다 (p=0.034). 아스피린 과민성 천식 환자군과 아스피린 내성 천식 환자군에서 CRTH2 -466T>C 유전자 다형성의 대립유전자 빈도의 차이가 통계적으로 유의하게 관찰되었다 (p<0.05). 또한, 아스피린 과민성 천식 환자군에서, CRTH2 -466TT 유전자형을 가진 환자들의 경우 -466CT 또는 -466CC 유전자형을 가진 환자들 보다 혈청 eotaxin-2 생성이 높게 관찰되었다. 인간 폐 상피세포를 이용한 시험관내 기능 연구를 통해 -466T 대립유전자는 낮은 luciferase activity를 보였고(p<0.001), 낮은 mRNA 발현을 보이고, 반면 높은 eotaxin-2 생성이 관찰되었다(p=0.003). EMSA 실험을 통해, -466T 대립유전자에 -466C 대립유전자보다 더 높은 친화도를 가지고 결합하는 전사인자를 확인하였다.
TBXA2R -4684T>C 유전자 다형성의 대립유전자 빈도의 차이는 아스피린 과민성 급성 두드러기 환자군과 정상 대조군에서 통계적으로 유의하게 관찰되었다 (p= 0.015). 아스피린 과민성 급성 두드러기 환자군에서, TBXA2R -4684T>C 유전자 다형성은 아토피와 연관성(p= 0.013)을 보였지만, 아스피린 과민성 만성 두드러기 환자군에서는 아토피와 연관성을 보이지 않았다. 인간 비반 세포주인 HMC-1을 이용한 시험관내 기능 연구를 통해 TBXA2R -4684T 대립유전자는 낮은 luciferase activity를 보였고, EMSA 실험을 통해 -4684T 대립유전자에 -4684C 대립유전자보다 더 높은 친화도를 가지고 결합하는 전사인자를 확인하였다. 또한, 아스피린 과민성 급성 두드러기 환자들은 정상 대조군에 비해 낮은 thromboxane B2 생성을 보였다 (p=0.016).
결론: 본 연구를 통해, 프로스타글란딘 D2 수용체, CRTH2,와 프로스타노이드 수용체, TBXA2R 유전자들의 유전자 다형성들이 아스피린 과민성과 연관성이 있으며, 특히 CRTH2 유전자는 아스피린 과민성 천식과 TBXA2R 유전자는 아스피린 과민성 급성 두드러기와 연관성이 있음을 확인하였다.
핵심어: 아스피린 과민성, 천식, 급성 두드러기, 유전자 다형성, 호산구TABLE OF CONTENTS
ACKNOWLEDGEMENT I
ABSTRACT II
TABLE OF CONTENTS VI
LIST OF FIGURES IX
LIST OF TABLES XI
ABBREVIATIONS XII
PART-I 1
GENETIC POLYMORPHISM OF CRTH2 IN ASPIRIN INTOLERANT ASTHMA 1
I.INTRODUCTION 2
II. MATERIALS AND METHODS 5
A. Study subjects and phenotyping 5
B. SNP identification and genotyping 6
C. Statistical analysis 7
D. In vivo functional study of CRTH2 -466T>C by measurement of eotaxin-1 and eotaxin-2 using enzyme-linked immunosorbent assay 7
E. Preparation of the CRTH2 construct and dual luciferase assay 8
F. Nuclear extract preparation and electrophoretic mobility shift assay 10
G. Preparation of CRTH2-coding sequence (CDS) (-466T or -466C) and in vitro functional study of eotaxin-2 production in A549 cells and mRNA expression 11
III. RESULTS 13
A. Clinical characteristics of the CRTH2 study subjects 13
B. Genotype and allele frequencies of the CRTH2 gene with the AIA phenotype 15
C. Eotaxin level according to the CRTH2 gene polymorphism 18
D. Eotaxin-2 level according to severity of AIA patients based on classification of FEV1 (%) 18
E. Endogenous expression of CRTH2 from various cell lines 20
F. Dual luciferase activity of CRTH2 -466T>C polymorphism 21
G. Effects of CRTH2 -466T>C gene polymorphisms on
transcriptional activity 22
H. CRTH2 expression and in vitro production of eotaxin-2 from CRTH2 transfected A549 cells 23
IV. DISCUSSION 31
PART-II 35
GENETIC POLYMORPHISM OF TBXA2R IN ASPIRIN INTOLERANT ACUTE URTICARIA 35
I. INTRODUCTION 36
II. MATERIALS AND METHODS 39
A. Study subjects and phenotyping 39
B. SNP identification and genotyping 40
C. Statistical analysis 40
D. Preparation of the TBXA2R construct and dual luciferase assay 41
E. Electrophoretic mobility shift assay 42
F. In vivo measurement of TXB2 production from sera of AIAU and NC 43
III. RESULTS 45
A. Clinical characteristics of the TBXA2R study subjects in AIAU 45
B. Genotype and allele frequencies of the TBXA2R gene with the AIAU phenotype 45
C. Dual luciferase activity of TBXA2R -4684T>C polymorphism 50
D. TBXA2R -4684T>C gene polymorphisms on transcriptional activity 50
E. TXB2 production according to TBXA2R -4684T>C gene polymorphism 56
IV. DISCUSSION 57
V. CONCLUSION 60
REFERENCES 61
국문요약 70Master- ABSTRACT -
Genetic Polymorphism of Prostanoid Receptors, CRTH2 and
TBXA2R in Aspirin Hypersensitivity
Background and Objective: Aspirin ingestion can induce a wide range of clinically recognized allergic reactions, including aspirin intolerant asthma (AIA), also called aspirin exacerbated respiratory disease (AERD), aspirin-intolerant urticaria/angioedema (AIU), chronic rhinitis, and anaphylaxis. We studied two major aspirin hypersensitivity, AIA and AIAU in an association with prostanoids receptor CRTH2 and TBXA2R. The human CRTH2 gene (official name GPR44) encodes a G protein-coupled chemoattractant receptor molecule which is expressed on Th2 cells including other allergy related cells like eosinophils, basophils and monocytes. Its importance in inflammatory cell activation and recruitment in vitro and in vivo has been continuously reported in several studies. Considering the fact that eosinophilic infiltration is more pronounced in AIA patients than it is in ATA patients, we hypothesized that activation of eosinophils via dysregulation of the CRTH2 gene may play an important role in AIA and may be an important marker of this entity. Therefore, this case control study was designed to determine if variation in the CRTH2 gene confers elevated risk for developing AIA.
Human Thromboxane A2 (TBXA2), is another eicosanoid product induced by cyclooxygenase may be as important mediator in aspirin hypersensitivity as it can induce bronchoconstriction and bronchohyperresponsiveness. TBXA2 exerts its action by interacting with the G protein-coupled thromboxane A2 receptor (TBXA2R).
Based on previous association of TBXA2R with asthma and atopy, we investigated whether genetic polymorphisms of the TBXA2R gene are associated with AIAU phenotype through a case-control of three study subjects.
Materials and Methods: The three studies groups 107 patients with AIA, 115 patients with ATA, and 133 normal healthy controls (NC) were recruited from Ajou University Hospital, Suwon, Korea. We genotyped two polymorphisms of CRTH2 located in front of exon-2 (upstream of the ATG) using a primer extension method and the SNAPshot ddNTP primer extension kit. (Applied Biosystems, Foster City, CA, USA). In case of TBXA2R gene polymorphism, three subject groups (167 patients with AIAU, 316 patients with AICU, and 265 patients with normal controls) were enrolled at Ajou University Hospital in Suwon, Korea. Two polymorphisms of TBXA2R (-4684T>C and 795 T>C) were genotyped using a primer extension method and the SNAPshot ddNTP primer extension kit (Applied Biosystems, Foster City, CA, USA). The functional effect of polymorphism of CRTH2 and TBXA2R were analyzed by luciferase reporter assay, electrophoretic mobility shift assay and ELISA.
Results: AIA patients had significantly higher serum eotaxin-2 levels than did those with ATA (p=0.034). A significant difference in the genotype frequencies of CRTH2 -466T>C was detected between AIA and ATA patients (p<0.05). The serum eotaxin-2 level was significantly higher in AIA patients carrying the TT genotype than those with the CT and CC type (p<0.05). In vitro functional study demonstrated that the -466T allele had lower luciferase activity (p<0.001) and lower mRNA expression with higher production of eotaxin-2 (p=0.003) in human lung epithelial cells. EMSA showed that CRTH2 -466T produced a specific band with a higher affinity than CRTH2 -466C had. AIA patients exhibited a significantly higher prevalence of paranasal sinusitis and nasal polyps compared to ATA patients (p<0.001 and p<0.001, respectively). The serum eotaxin-2 level was significantly higher in AIA patients than in ATA patients (p=0.034) in our study, while no difference was noted in the eotaxin-1 level.
Furthermore, the genotype frequency of TBXA2R -4684T>C was significantly different AIAU and NC on both co-dominant and recessive analysis models. Specifically, AIAU patients showed a significantly higher frequency of the homozygous TT genotype of TBXA2R -4684T>C compared to NC (p=0.015). In addition, there was significant association of atopy status according to TBXA2R -4684T>C polymorphism. AIAU patients carrying TT genotype showed significantly higher atopy status compared to CT or CC type (p=0.013) but not in AICU patients (P>0.05). In vitro functional study demonstrated that the -4684T allele had lower luciferase activity (p<0.001) in HMC-1 cells. EMSA showed that TBXA2R -4684T produced a specific band with a higher affinity than TBXA2R -4684C. There was a significant difference between AIAU vs. NC with respect to serum Thromboxane B2 production (TXB2). AIAU patients showed a significantly lower TXB2 production compared to patients with NC (p=0.016).
Conclusion: These study suggest that genetic variability of prostanoid receptors CRTH2 and TBXA2R may be associated with aspirin hypersensitivity, in particular CRTH2 ？466T allele showed higher frequency and increased serum and cellular eotaxin-2 production through lowering of CRTH2 expression in AIA and TBXA2R -4684T allele showed higher frequency and lowered TXB2 production through lowering of TBXA2R expression in AIAU.
__________________________________________________________________________
Key words: aspirin hypersensitivity, respiratory disease, acute urticaria, genetic polymorphism, eosinophils, eotaxin, thromboxane
The Roles of Serum Amyloid A-1 and Tissue Inhibitor of Metalloproteinase-1 in Asthma Pathogenesis
No full text학위논문(박사)--생명과학과,2023. 8배경: 천식은 가역성 기도 폐쇄를 특징으로 하는 만성 기도 염증성 질환이다. 이 중 5%-10%를 차지하는 중증 천식(SA)은 지속적인 기도 염증을 나타내며, ICS를 포함한 약물 치료에도 폐기능이 감소한다. Serum amyloid A (SAA)은 호중구성 천식(NA, 가래 호중구 수 65%)의 바이오마커 중 하나로 1초간 강제 호기량 감소 (FEV1%) 감소와 관련성이 보고되었다. 반면, tissue inhibitor of metalloproteinase-1 (TIMP1)은 TIMP 계열이며, 세포외 조직에서 주로 matrix metalloproteinases-9 (MMP9)의 억제제로 알려져 있고, 최근 TIMP1와 MMP9는 상호 작용을 통하여 기도 개형을 증가시킨다. SAA1과 TIMP1는 주로 기도 상피 세포 (AECs)에서 다양한 자극에 의해 분비되며, SA의 중증도에 영향을 미친다.
목적: 본 연구는 (1) 천식 표현형(NA vs non-NA)에 따른 혈청 SAA1의 임상적 유용성, (2) 외부/내부 자극에 의한AEC로부터 SAA1 생성, (3) 호중구/대식세포의 활성화에 대한 SAA1의 효과, (4) 천식 마우스 모델에서 SAA1의 효과, (5) SA 표현형(2형 대 비 2형)에 따른 임상적 매개변수와 관련한 혈청 TIMP1의 임상적 유의성, (6) 다양한 자극에 의한 AECs의 TIMP1 생성, (7) 호산구/대식세포의 활성에 의한 TIMP1의 효과, (8) TIMP1 및 항 TIMP1 항체가 생체 내 기도 염증/개형에 미치는 영향, 총 8가지를 확인하고자 하였다.
재료 및 방법: 첫번째 연구에서는, 122명의 천식 환자와 60명의 건강한 대조군(HCs)에서 혈청 SAA1를 측정하였다. 그리고 AECs에서 SAA1의 생산과 대식세포 및 호중구에 미치는 영향을 시험관 내 및 천식 마우스 모델에서 관찰하였다. 두 번째 연구에서는, 성인 천식 환자 250명(SA군, 54명; non-SA군, 196명)과 140명의 HCs 에서 혈청 TIMP1치를 측정하였다. 호산구 및 대식세포의 활성화에 대한 TIMP1의 효과와 AECs로부터 다양한 외부 자극에 따른 TIMP1의 방출 정도를 시험관내 및 생체내에서 평가하였다.
결과: 혈청 SAA1 수치는 천식 환자에서 HCs보다 유의하게 높았고(P = 0.014), 천식 환자들 중 호중구성 천식 환자군이 비 호산구성 천식군보다 유의하게 높았다 (P < 0.001). Poly I-C 처리 후 AECs에서 SAA1 생산이 현저하게 증가하였으며, 이는 호중구 처리 후 더욱 증가되었다. SAA1 처리 후, AECs에서 IL6, IL8 및 S100 칼슘 결합 단백질 A9 유리가 증가하였다. 또한, SAA1은 호중구에서 천식 환자의 말초 혈액에서 호중구 및 대식세포를 활성화하고, 호중구 세포외 트랩 형성 및 염증성 사이토카인 생산을 증가시켰다. 알부민으로 유발한 천식 마우스에서 poly I-C 자극은 기관지 폐포 세척액(BALF)에서 SAA1 수치과 IL17A/인터페론-감마/IL33 수치을 유의하게 증가시켰다. NA 마우스 모델의 BALF와 혈청에서 가장 높은 수치의 SAA1과 호중구 증가증이 나타났고, 혼합 과립구성 천식 마우스 모델이 그 뒤를 이었다. 특히, SAA1은 천식 마우스의 활성화된 CD4+ T 림프구로부터 RORt 발현을 유도하였다.
두 번째 연구에서, 혈청 TIMP1 수치는 천식 환자에서 HCs에 비해 유의하게 높았으며, 천식 환자중에서, non-SA군보다 SA군에서, 비2형군보다 2형 SA군에서 유의하게 높았다(각 P <0.01). SA 환자군에서 혈청 TIMP1과 FEV1 % 값 사이의 음의 상관관계(r = -0.400, P = 0.003)가 관찰되었다. 시험관 내 연구결과로 TIMP1는 AECs에서 다양한 외부 자극인 poly I-C, IL13, 호산구 세포외 트랩(EET) 및 호산구와 공동 배양 후 방출되었다. 동물 모델에서는 TIMP1 자극 후 호산구성 기도 염증이 관찰되었으며, 이는 스테로이드 치료로 완전히 억제되지 않았다. 또한, TIMP1은 호산구와 대식세포를 직접 활성화하고, EET와 대식세포의 방출을 유도하여, 항-TIMP1 항체 치료후 M2로의 분극화가 억제되었다.
결론: 이러한 결과는 SAA1이 NET 형성, M1 방향 대식세포 활성화 및 T 헬퍼(Th)2/Th17 우세 세포와 함께 호중구를 활성화 및 호중구 기도 염증을 유발하여, NA 또는 MA의 표현형에 기여한다. TIMP1은 SA 환자에서 호산구성 기도 염증을 강화하며, 혈청 TIMP1은 2형 SA에 대한 잠재적인 바이오마커 및/또는 치료 표적이 될 수 있다.CHAPTER-I 1
Serum Amyloid A-1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients 1
I. INTRODUCTION 2
II. MATERIALS AND METHODS 4
A. Study subjects 4
B. Human AEC culture and stimulation 4
C. Peripheral blood neutrophil (PBN) and monocyte isolation and culture 5
D. Neutrophil extracellular trap (NET) induction 5
E. Reactive oxygen species (ROS) quantification 6
F. Neutrophil migration assay 6
G. Macrophage polarization and stimulation 6
H. Asthma mouse models 7
I. Measurement of airway hyperresponsiveness (AHR) 8
J. Differential cell count 9
K. Murine CD4+ T cell isolation and stimulation 9
L. Human T lymphocyte cell line culture and stimulation 9
M. ELISA 9
N. Western blot analysis 10
O. Immunofluorescence with confocal microscope 10
P. Statistical analysis 10
III. RESULTS 12
A. Clinical characteristics of the study subjects 12
B. Factors for inducing SAA1 production from AECs 16
C. Effect of SAA1 on neutrophil activation 18
D. Effect of SAA1 on macrophage activation 20
E. SAA1 expression in allergic asthma mice infected by virus 22
F. SAA1 expression in the mouse model of asthma phenotypes 24
G. Effect of SAA1 on Th2/Th17 cytokine expression from splenic CD4+ T cells of allergic asthma mice 26
H. Effect of SAA1 on Th2 cytokine releases from Jurkat T cell line 27
IV. DISCUSSION 28
CHAPTER-II 32
Tissue Inhibitor of Metalloproteinase-1 Enhances Eosinophilic Airway Inflammation in Severe Asthma 32
I. INTRODUCTION 33
II. MATERIALS AND METHODS 35
A. Study subjects 35
B. Human AEC culture and stimulation 35
C. Peripheral blood eosinophil (PBE) and monocyte stimulation 36
D. Eosinophil migration assay 37
E. Human mast cell line culture and stimulation 37
F. Human macrophage culture and stimulation 38
G. Mouse model 38
H. ELISA 40
I. Western blot analysis 40
J. Flow cytometry 40
K. Immunofluorescence with confocal microscope 42
L. Statistical analysis 43
III. RESULTS 44
A. Clinical characteristics and serum cytokine levels of the study subjects 44
B. Clinical characteristics of the asthmatic subjects according to serum TIMP1 level 48
C. TIMP1 and MMP9 production from human AECs 51
D. Effect of TIMP1 on eosinophil migration and activation 54
E. Effect of TIMP1 on LAD2 cells 56
F. Effect of TIMP1 on human macrophages 57
G. Effect of TIMP1 on airway inflammation in vivo 59
H. Effect of anti-TIMP1 antibody on airway inflammation of chronic allergic asthma mouse model 62
IV. DISCUSSION 66
CONCLUSION 69DoctorBackground: Neutrophilia (sputum neutrophils 65%) in the sputum and its mediators are attributed to neutrophilic airway inflammation that is associated with poor steroid responsiveness and severe asthma (SA). Although many researchers have investigated its underlying mechanisms, potential biomarkers and therapeutics, it has not been completely understood. Serum amyloid A (SAA) was proposed as a biomarker for neutrophilic asthma (NA) and was associated with impaired lung function. SAA1, a primary precursor of SAA, was involved in lipid metabolism, bacterial clearance and inflammatory regulations; nevertheless, its clinical significance and function in NA have not been studied. Furthermore, the presence of eosinophilia in the blood/sputum of severe asthmatic patients (type 2 SA) contributed to recurrent asthma exacerbations and low lung function even on maintenance medication including inhaled corticosteroids (ICS) and long-acting beta-agonist (LABA). In addition, tissue inhibitor of metalloproteinase-1 (TIMP1) is a member of the TIMP family and play the role of a major inhibitor of metalloproteinase-9 (MMP9) in extracellular tissues. The pathogenic mechanism of TIMP1 on airway inflammation and remodeling remained uncertain, but accumulating evidence has shown that TIMP1 and MMP9 co-work in airways to upregulate tissue remodeling.
Objective: This thesis sought to investigate: 1) clinical relevance of serum SAA1 according to asthma phenotype (NA vs non-NA); 2) SAA1 production from airway epithelial cells (AECs) upon external/internal stimuli; 3) the effect of SAA1 on the activation of neutrophils/macrophages; 4) SAA1 expression in different asthma mouse models; 5) clinical relevance of serum TIMP1 level according to SA phenotype (type 2 SA vs non-type 2 SA); 6) TIMP1 production from AECs upon various stimuli; 7) the effect of TIMP1 on the activation of eosinophils/macrophages; and 8) the effects of TIMP1 and anti-TIMP1 antibody on airway inflammation and remodeling in vivo.
Methods: In the first study, 122 adult asthmatics (78 patients with NA and 44 those with non-NA) and 60 healthy controls (HCs) were enrolled to measure serum SAA1 levels. The production of SAA1 from AECs and its effects on neutrophils and macrophages were investigated.
In the second study, we enrolled 250 adult asthmatics (54 patients with SA and 196 those with non-SA) and 140 HCs to measure serum TIMP1 levels. The release of TIMP1 from AECs and its effects on the activations of eosinophils and macrophages were evaluated.
Results: Serum SAA1 levels were significantly higher in asthmatic patients than in HCs (P = 0.014); among asthmatics, patients with NA showed significantly higher SAA1 levels than those with non-NA (P < 0.001). In vitro investigations showed that poly I-C stimulation markedly enhanced the production of SAA1 from AECs, which was further augmented by neutrophils; SAA1 could induce the production of IL6, IL8 and S100 calcium-binding protein A9 from AECs. Additionally, SAA1 activated peripheral neutrophils and monocyte-derived macrophages from asthmatics to induce the neutrophil extracellular trap (NET) formation and pro-inflammatory cytokine production. In ovalbumin (OVA)-induced allergic asthma mice, poly I-C stimulation significantly increased SAA1, IL17A, interferon-gamma and IL33 levels in the bronchoalveolar lavage fluid (BALF). The highest levels of SAA1 were noted in the BALF and sera of the NA mouse model, followed by the mixed granulocytic asthma model. Especially, SAA1 induced IL17 and retinoic acid receptor-related orphan receptor gamma t expressions from activated CD4+ T lymphocytes in OVA-induced allergic asthma mice.
In the second study, significantly higher levels of serum TIMP1 were noted in asthmatics than in HCs, in the SA group than in the non-SA group and in the type 2 SA group than in the non-type 2 SA group (P < 0.01 for all). A negative correlation between serum TIMP1 and FEV1% values (r = –0.400, P = 0.003) was noted in the SA group. In vitro investigations demonstrated that TIMP1 was released from AECs in response to poly I-C, IL13, eosinophil extracellular traps (EET) and in coculture with eosinophils. TIMP1-stimulated mice showed eosinophilic airway inflammation, which was not completely suppressed by steroid treatment. In vitro and in vivo functional investigations showed that TIMP1 directly activated eosinophils and macrophages, and induced the release of EET and macrophages polarization toward M2 subset, which was suppressed by anti-TIMP1 antibody.
Conclusions: These findings suggest that SAA1 may cause neutrophilic airway inflammation in adult asthmatic patients by activating neutrophils to form NET, macrophages to polarize toward the M1 subset, and CD4+ T cells to polarize toward T helper (Th) 2/Th17 predominant cells, resulting in steroid resistance. We propose SAA1 as the potential biomarker for the prediction of NA phenotype. In patients with SA, TIMP1 may contribute to the upregulation of eosinophilic airway inflammation and remodeling via migrating eosinophil into the airways and subsequently activating eosinophils to release EET, which further activate group 2 innate lymphoid cells. In the chronic allergic asthma mouse model, blocking TIMP1 with neutralizing antibodies reduced airway hyperresponsiveness, eosinophilic airway inflammation and remodeling. Taken together, modulating TIMP1 and EET production may be a potential treatment strategy for type 2 SA
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Various Clinical Phenotypes and Approach to Patients with NSAID Hypersensitivity
No full textNonsteroidal anti-inflammatory drugs (NSAIDs) can induce hypersensitivity reactions with various clinical manifestations including acute/delayed reactions and three common phenotypes, NSAID/aspirin-exacerbated respiratory disease (NERD or AERD) with/without chronic rhinosinusitis (CRS) and nasal polyps, NSAID-exacerbated cutaneous disease (NECD). NSAID-induced urticaria/angioedema (NIUA). NIAU is commonly combined with anaphylaxis and named as NIUAA. The major pathogenic mechanism is the inhibition of cyclooxygenase-1 with a reduction in prostaglandin E2 levels, leading to the overproduction of cysteinyl leukotrienes and activation of inflammatory cells, including eosinophils and mast cells. To confirm the diagnosis, provocation testing via the oral route or inhalation remains the gold standard; in vitro diagnostic methods are still not available. Essential managements are: (1) avoidance of cross-reacting NSAIDs along with the use of alternative analgesics; (2) pharmacologic treatment should follow standard guidelines in patients with underlying asthma/rhinitis (NERD, CRS), and urticaria (NECD). Aspirin desensitization and biologic treatment can be done when indicated. Delayed reactions, including fixed drug eruptions, maculopapular eruptions, and severe cutaneous adverse reactions, are rare and mediated by T-cell responses. Symptomatic treatment with avoidance is essential. NSAID is a cofactor of food-dependent exercise-induced anaphylaxis. In this talk, we will update various clinical phenotypes and discuss practical issues faced in real clinical practice
- …
