Genetic effect of <i>CCR3</i> and <i>IL5RA</i> gene polymorphisms on eosinophilia in asthmatic patients

Lee, June-Hyuk,Chang, Hun Soo,Kim, Ji Hyun,Park, Se-Min,Lee, Yong Mok,Uh, Soo Taek,Rhim, Taiyoun,Chung, Il Yup,Kim, Yong-Hoon,Park, Byung Lae,Park, Choon-Sik,Shin, Hyoung Doo Elsevier 2007 The journal of allergy and clinical immunology Vol.120 No.5

<P><B>Background</B></P><P>Eosinophilic infiltration and peripheral blood eosinophilia in asthma require the cooperation of eosinophil-specific cytokines and chemokines and their receptors.</P><P><B>Objective</B></P><P>We investigated the association of polymorphisms in <I>CCR3</I> and <I>IL5RA</I> with asthma susceptibility or peripheral blood eosinophilia and the effects of the polymorphisms on receptor expression.</P><P><B>Methods</B></P><P>Polymorphisms in <I>CCR3</I> and <I>IL5RA</I> were identified and genotyped in 576 asthmatic patients and 180 healthy control subjects. CCR3 and IL-5 receptor α (IL-5Rα) protein expression on eosinophils was measured by means of flow cytometry.</P><P><B>Results</B></P><P>Although polymorphisms in <I>CCR3</I> were not associated with asthma susceptibility, the <I>CCR3</I> haplotype <I>ht2</I> showed a negative gene dose effect on the eosinophil count (<I>P</I> = .003–.009). <I>IL5RA c.−5091G>A</I> was weakly associated with eosinophil count. The effects of <I>ht2</I> were greater when paired with <I>IL5RA c.−5091A</I> (<I>P</I> = .001–.002). CCR3 protein expression was higher on eosinophils of asthmatic patients without <I>ht2</I> than in those with <I>ht2</I>. Asthmatic patients with the <I>IL5RA c.−5091A</I> allele showed higher IL-5Rα expression than those who were homozygous for the G allele.</P><P><B>Conclusion</B></P><P>The genetic association between <I>CCR3</I> polymorphisms and the number of circulating eosinophils was revealed as a novel finding. These associations were more pronounced when the <I>CCR3</I> polymorphisms were paired with polymorphisms in <I>IL5RA</I>. The protein expression levels of CCR3 and IL-5Rα on peripheral blood eosinophils are associated with the polymorphisms on their own genes.</P><P><B>Clinical implications</B></P><P>The identification of single nucleotide polymorphisms and haplotypes of <I>CCR3</I> and <I>IL5RA</I> might be useful in developing markers for intermediate phenotypes of eosinophil number and in designing strategies to control diseases related to hypereosinophilia.</P>

DBA/1JCrj Mouse에 있어서 콜라젠유도관절염에 관한 면역학적 고찰

박승규,이지연,정일엽,최용경,최인성,김효준 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.3

There is growing evidence that a variety of cytokines are secreted by cells at the inflammation sites of rheumatoid arthritis (RA) and that CDS+ B cell, a minor subtype of B cell population producing natural autoantibodies, is implicated in pathogenesis of RA. In this study, we evaluated the both cytokne levels and change of CDS+ B cell population in the peripheral blood from DBA/lJCrj mice(H-2`) which are collagen-induced arthritis(CIA) susceptible strain. Surprisingly, the healthy DBA/1JCrj and MRL/Ipr/Ipr(H-2'`) mice which were autoimmune susceptible strains tested in this experiment, showed lower IL-4, IL-6 and IL-10 levels in serum by 60% than heal-thy normal mouse strains such as Balb/c(H-2d), C57BL/6(H-2') and outbred ICR mice. MRL/lpr/ 1pr mice in which onset of spontaneous autoimmune disease is dependent upon age were similar to healthy DBA/1JCrj mice in levels of the cytokines in the serum when they are young. After the CIA(for DBA/1JCrj) or the spontaneous autoimmune disease(for MRL/lpr/Ipr) had been developed in the susceptible mouse strains, the levels of IL-6 and IL-4 in the serum were increased to 1.8- and 13-fold, respectively, as compaired with those from control groups while level of IL-10 remained relatively constant. The elevated levels of IL-4- and IL-6, however, in the serum from mice with disease status were still below those of the healthy normal mouse strains. On the other hand, CDS+ B cell population in the peripheral blood, which were reported to be increased with the development of RA for human, was rather significantly decreased for the CIA-induced DBA/lJCrj mice as evidenced by FACS analysis. It could be due to the differences in the pathogenic mechanism between CIA and RA. Taken together, our results. suggest that the levels of both these cytokines and CDS+ B cells may be utilized as important diagnostic markers for arthritides.

Interferon-γ Inhibits in vitro Mobilization of Eosinophils by Interleukin-5

Park, Choon-Sik,Choi, Eun Nam,Kim, Jung Sun,Choi, Yun Sung,Rhim, Tai Youn,Chang, Hun Soo,Chung, Il Yup S. Karger AG 2005 International archives of allergy and immunology Vol.136 No.3

<P><I>Background:</I> Th2 cytokines play pivotal roles in allergic inflammation, including eosinophilia, and their actions are antagonized by Th1 cytokines, conferring them therapeutic potential. <I>Methods:</I> In this study, we examined the ability of a number of cytokines to suppress the activation of eosinophils that function as effector cells for allergic airway diseases. <I>Results:</I> Interleukin (IL)-5, IL-6, and tumor necrosis factor (TNF) induced an eosinophil shape change, whereas interferon (IFN)-γ significantly inhibited the shape change. Other cytokines, including IL-1β, IL-4, IL-10 and IL-13, had little or only slightly enhancing or reducing effects on the shape change. We further analyzed the IFN-γ effect, showing that pretreatment with IFN-γ strongly suppressed IL-5-induced eosinophil shape change, and cycloheximide (CHX) abrogated the suppression by IFN-γ, suggesting that new protein synthesis is required for the inhibitory effect by this cytokine. In agreement with these results, IFN-γ blocked the eosinophil migration and ERK phophorylation induced by IL-5, and the addition of CHX restored eosinophil chemotaxis. <I>Conclusions:</I> Collectively, IFN-γ may attenuate eosinophilic inflammation by directly negating eosinophil mobilization.</P><P>Copyright © 2005 S. Karger AG, Basel</P>

Cell Surface Expression of Tumor Necrosis Factor - Alpha by Activated Rat Astrocytes

Chung, Il Yup,Benveniste, Etty N 생화학분자생물학회 1982 BMB Reports Vol.29 No.6

Astrocyte are the major glial cell type in the central nervous system (CNS), and analogous to macrophage, mediates the number of immune responses such as production of cytokines including tumor necrosis factor alpha (TNF-a) upon activation. TNF-a has been implicated in neuroimmunological disorders through killing oligodendrocytes and thus causing demyelination. It has been previously demonstrated that mitogen-activated T cells synthesized a 26 kDa precursor form of TNF-α which is bound to the surface of a membrane, and is later secreted as a 17 kDa mature version. In order to examine whether astrocytes would produce the transmembrane form of TNF-α, astrocytes were stimulated with biological stimuli and the membrane form of TNF-α was analyzed by Western blest and FRCS analysis. When astrocytes are stimulated with lipopolysaccharide (LPS), IFN-γ/LPS, or IFN-γ/IL-1β, they were able to express a membraneanchored TNF-α of approximately 26 kDa protein which was immunoreactive to an anti-TNF-α antibody. whereas unstimulated astrocytes or astrocytes treated with IFN-γ or IL-1β alone was not. Our FRCS data were also consistent with the immunoblot analysis. Our result suggests that the membrane form of TNF-α expressed by activated astrucytes may cause local damage to oligodendrocytes by direct cell-cell contact and contribute to demyelination observed in multiple sclerosis (MS) and experimental allergic encephalomyelitis (ERE).

Centipede Scolopendra subspinipes multilans L. Koch로부터 항균력이 있는 Centipedin의 추출, 정제 및 특성연구

김기태,최진성,민천홍,정일엽,박경배,조기승 한양대학교 이학기술연구소 1999 이학기술연구지 Vol.1 No.-

항균력이 있는 화합물인 Centipeding을 지네 Scolopendra subspinipes multilans L. Koch로부터 지용성 유기용매로 추출하였고, silicic acid column과 high S cation exchange chromatography 및 reverse-phase HPLC 등을 사용하여 고순도로 정제하였다. Gram-positive bacteria인 Staphylococcus aureus ATCC 6538P 및 gram-negative인 Klebsiella pneumoniae ATCC 8308 균주에 대하여 강한 항균력을 나타내었다. Dragendorff 시약에 의한 발색반응을 시행한 결과 Centipedin의 aromatic ring에는 nitrogen이 없음을 확이하였고 IR, Mass, NMR 분석 등에 의하여 Centipedin의 분자량은 162.1, 화학적 구조는 hydroxyl기와 carbonyl기를 함유하는 -hydroxyisocumarin으로 확인되었다. UV-spectrum 검색결과에 의하면 최대 흡수파장이 243 ㎚임을 확인하였고, melting point는 122℃이었으며, 실온에서 3개월간 방치된 Centipedin이 여전히 항균활성 및 화학적 구조를 유지하여 물리화학적 및 생물학적으로 안정한 화합물임으 알 수 있었다. The compound which has antibiotic activity was extracted and purified from centipede Scolopendra subspinipes multilans L. Koch by diethyl ether, silicic acid column and high S cation exchange chromatography followed by reverse-phase HPLC. This compoung named "Centipedin" showed antibiotic activity aganist microorganisms such as Gram^(+) -bacteria Staphylococcus aureus ATCC 6538P and Gram^(-) -bacteria Klebsiella pneumoniae ATCC 8308. Dragendorff test showed that there is no aromatic nitrogen within the Centipedim. The molecular weight of the Centipedin was 162.1 daltons as determined by mass spectrometry. The instrumental analysis of IR and NMR techniques revealed the molecular structure of Centipedim as 8-hydroxyisocoumarin. The UV-spectrum of Centipedin showed maximum absorption at 243 ㎚, and melting point was 122℃. Centipedin was stable at room temperature for three months in appropriate buffers.

Inflammatory mediators ATP and S100A12 activate the NLRP3 inflammasome to induce MUC5AC production in airway epithelial cells

Kim, Karam,Kim, Hye Jeong,Binas, Bert,Kang, Jin Hyun,Chung, Il Yup Elsevier 2018 Biochemical and biophysical research communication Vol.503 No.2

<P><B>Abstract</B></P> <P>Danger-associated molecular patterns (DAMPs) play a proinflammatory role in the pathogenesis of airway obstructive diseases such as severe asthma and chronic obstructive pulmonary disease. The NLRP3 inflammasome is a cytosolic multiprotein platform that activates the caspase-1 pathway in response to inflammatory stimuli such as DAMPs. ATP and S100 proteins are newly identified DAMPs that accumulate in inflamed airways. We previously demonstrated that S100A8, S100A9, and S100A12 induce production and secretion of MUC5AC, a major mucin in the conducting airway mucosa. The purpose of this study was to determine the involvement of NLRP3 inflammasome in, and the contribution of ATP to, S100 protein-induced MUC5AC production by NCI-H292 mucoepidermoid carcinoma cells. Stimulation with either S100A12 or ATP led to MUC5AC production at comparable levels. Simultaneous treatment with both stimuli resulted in additive increases in NLRP3, active caspase-1, IL-1β, NLRP3/caspase-1 colocalization, and MUC5AC. NLRP3 siRNA or inhibitors of NF-κB, NLRP3 inflammasome oligomerization, or caspase-1 nearly completely inhibited ATP- and S100A12-mediated MUC5AC production. Furthermore, S100A12-as well as ATP-mediated MUC5AC production was almost equally blunted by both nonspecific and specific antagonists of the purinergic receptor P2X7, a principal receptor mediating NLRP3 inflammasome activation by ATP. Thus, these two danger signals contribute to MUC5AC production in airway epithelial cells through overlapping signaling pathways for NLRP3 inflammasome activation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> S100A12 activates NLPR3 inflammasomes to induce MUC5AC production in epithelial cells. </LI> <LI> ATP induces MUC5AC production in a mechanistically similar mode to S100A12. </LI> <LI> S100A12-mediated MUC5AC production involves engagement of ATP. </LI> </UL> </P>

Cell Surface Expression of Tumor Necrosis Factor-Alpha by Activated Rat Astrocytes

Chung, Il-Yup,Benveniste, Etty N. Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.6

Astrocyte are the major glial cell type in the central nervous system (CNS), and analogous to macrophage, mediates the number of immune responses such as production of cytokines including tumor necrosis factor alpha ($TNF-{\alpha}$) upon activation. $TNF-{\alpha}$ has been implicated in neuroimmunological disorders through killing oligodendrocytes and thus causing demyelination. It has been previously demonstrated that mitogen-activated T cells synthesized a 26 kDa precursor form of $TNF-{\alpha}$ which is bound to the surface of a membrane, and is later secreted as a 17 kDa mature version. In order to examine whether astrocytes would produce the transmembrane form of $TNF-{\alpha}$, astrocytes were stimulated with biological stimuli and the membrane form of $TNF-{\alpha}$ was analyzed by Western blot and FACS analysis. When astrocytes are stimulated with lipopolysaccharide (LPS), $IFN-{\gamma}/LPS$, or $IFN-{\gamma}/IL-1{\beta}$, they were able to express a membrane-anchored $TNF-{\alpha}$ of approximately 26 kDa protein which was immunoreactive to an $anti-TNF-{\alpha}$ antibody, whereas unstimulated astrocytes or astrocytes treated with $IFN-{\gamma}$ or $IL-1{\beta}$ alone was not. Our FACS data were also consistent with the immunoblot analysis. Our result suggests that the membrane form of $TNF-{\alpha}$ expressed by activated astrocytes may cause local damage to oligodendrocytes by direct cell-cell contact and contribute to demyelination observed in multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE).

The Purification and Characterization of Bacillus subtilis Tripeptidase(PepT)

Chung,Il Yup,Park,Yong Seek,Kim,Hyo Joon,Yong,Whan-Mi,Cha,Myung Hoon,Lee,Young Seek The Korea Science and Technology Center 1999 BMB Reports Vol.32 No.3

A tripeptidase (PepT) was purified to homogeneity from Bacillus subtilis through four sequential chromatographies including DEAE-Sepharose ion exchange, hydroxylapatite, mono-Q FPLC ion exchange, and Superose-12 FPLC gel filtration. The apparent molecular mass of the enzyme was 49,200 Da and 51,400 Da as determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, and the enzyme exists in a monomeric form. The physico-chemical properties of the enzyme were as follows: optimum pH at 7.5, optimum temperature at 60℃, and pI at 4.9. The Km and Vmax values of the enzyme were 4.3 mM and 2.5 mmol/min/mg, respectively, with Met-Ala-Ser as substrate. The B. subtilis PepT requires Co²+ ion(s) for activation, while it is inactivated by EDTA and 1,10-phenanthroline, suggesting that it is a metalloprotein. The enzyme was not inhibited by any of serine protease, aspartic protease, or leucine amino-peptidase inhibitors. The enzyme showed comparable activities towards four different substrates including Met-Ala-Ser, Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The amino terminal sequences of PepT determined by Edman degradation was found to be MKEEIIERFTTYVXV and turned out to be identical to that of PepT deduced from a cloned B. subtilis pepT.

Proinflammatory Cytokines과 TGF-beta가 섬유모세포의 증식에 미치는 영향

김철 ( Chul Kim ),박춘식 ( Choon Sik Park ),김미호 ( Mi Ho Kim ),장헌수 ( Hun Soo Chang ),정일엽 ( Il Yup Chung ),기신영 ( Shin Young Ki ),어수택 ( Soo Taek Uh ),문승혁 ( Seung Hyuk Moon ),김용훈 ( Yong Hoon Kim ),이희발 ( Hi Bal 대한결핵 및 호흡기학회 1998 Tuberculosis and Respiratory Diseases Vol.45 No.4

골수강내 금속정술과 금속판 내고정술을 이용한 상완골 간부 골절의 치료 결과 비교

정의섭,전일엽 대한골절학회 1999 대한골절학회지 Vol.12 No.3

The ideal treatment of acute fractures of the humeral shaft remains controversial. Recently, tendency of surgical treatment has been increased. Authors reviewed and analyzed plate fixation in 40 patients and intramedullary nailing in 26 patients having humeral shaft fractures. These patients were treated at the department of orthopaedic surgery, Chonju Presbyterian Medical Center from June 1994 to November 1997. There is no significant difference in union time, operation time, functional result. But, non-union rate of intramedullary nailing is superior to that of plate fixation. So, We concluded that plate fixation in humerus shaft fracture is more acceptable than intramedullary nailing, except severe osteoporosis, severe communited fracture, segmental fracture, pathologic fracture.