이질아메바에 의한 인체 대장상피세포주 HT-29에서의 interleukin-8 유전자의 발현

김정목,정현채,임경일,조양자,김정룡 INSTITUTE OF TROPICAL MEDICINE YONSEI UNIVERSITY 1995 YONSEI REPORTS ON TROPICAL MEDICINE Vol.26 No.1

이질아메바에 의한 장염 환자의 조직 또는 이질아메바를 실험적으로 감염시킨 동물의 조직 검사에서 호중구의 침윤이 특징적으로 관찰된다. 그러나 이와같은 호중구의 침윤을 설명할 수 있는 기전에 대한 연구는 매우 미흡하다. 따라서 본 연구자들은 아메바 감염 초기에 인체 대장상피세포에서 interleukin-8(IL-8)이 유도되어 호중구 침윤과 같은 염증반응이 유발될 것이라는 가설을 설정하였다. 이를 위하여 인체 대장상피세포주인 HT-29에 이질아메바 영양형을 실험적으로 노출시킨 뒤 발현되는 IL-8 mRNA를 역전사 중합효소법(reverse transcriptional polymerase chain reaction, RT-PCR)으로 검사함과 동시에 발현된 IL-8 mRNA를 인공적으로 합성시킨 표준 RNA와 RT-PCR법을 이용하여 정량하였다. 실험 결과 이질아메바 영양형에 노출된 30분 후 부터 IL-8 mRNA가 발현되기 시작하였다. 그리고 그 발현 분자수는 노출 시간의 증가에 따라 계속 증가하여 3시간 대에는 3.1×10(7) molecules/㎍ total RNA를 나타내었다. 동시에 IL-8 mRNA의 발현은 노출시킨 이질아메바 영양형의 수에 비례하였다. 즉, HT-29/아메바 영양형의 비율이 10:1인 경우 IL-8 mRNA의 발현 분자수는 1.2×10(7) molecules/㎍ total RNA로 나타났다. 이와같은 IL-8 mRNA의 발현은 IL-8 단백질 분비로 이어짐을 ELISA 검사로 확인할 수 있었다. 한편 이질아메바 파쇄액(lysate)도 대장상피세포주인 Caco-2에서 IL-8 mRNA발현을 유도하였다. 결론적으로 본 실험은 이질아메바 감염 초기에 대장상피세포로 부터 IL-8이 발현되며 이에 의하여 염증반응이 촉발될 가능성이 있음을 시사해 준다. The protozoan parasite, Entamoeba histolytica, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to E. histolytica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the finding that human epithelial cells produce the potent neutrophil chemoattractant and activator, interlukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to E. histolytica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to E. histolytica trophozoities for 30 minutes. 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polymerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1×10(7) molecules/㎍ of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA). Lysates of E. histolytica also induced expression of mRNA for IL-8 in colon epithelial cells. These results suggest that acute inflammatory reaction by E. hisstolytica may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

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>

Activation of Transient Receptor Potential Melastatin Family Member 8 (TRPM8) Receptors Induces Proinflammatory Cytokine Expressions in Bronchial Epithelial Cells

김주희,Young Sook Jang,Hwan Il Kim,박지영,박성훈,Yong Il Hwang,Seung Hun Jang,Ki-Suck Jung,Hae Sim Park,Choon-Sik Park 대한천식알레르기학회 2020 Allergy, Asthma & Immunology Research Vol.12 No.4

Purpose: Cold air is a major environmental factor that exacerbates asthma. Transient receptor potential melastatin family member 8 (TRPM8) is a cold-sensing channel expressed in the airway epithelium. However, its role in airway inflammation remains unknown. We investigated the role of TRPM8 in innate immune responses in bronchial epithelial cells and asthmatic subjects. Methods: The TRPM8 mRNA and protein expression on BEAS2B human bronchial epithelial cells was examined by real-time polymerase chain reaction (PCR), immunofluorescence staining and western blotting. Additionally, interleukin (IL)-4, IL-6, IL-8, IL-13, IL-25 and thymic stromal lymphopoietin (TSLP) levels before and after menthol, dexamethasone and N-(4-tert-butylphenyl)-4-(3-chloropyridin-2-yl) piperazine-1-carboxamide (BCTC) treatments were measured via real-time PCR. TRPM8 protein levels in the supernatants of induced sputum from asthmatic subjects and normal control subjects were measured using enzyme-linked immunosorbent assay, and mRNA levels in sputum cell lysates were measured using real-time PCR. Results: Treatment with up to 2 mM menthol dose-dependently increased TRPM8 mRNA and protein in BEAS2B cells compared to untreated cells (P < 0.001) and concomitantly increased IL-25 and TSLP mRNA (P < 0.05), but not IL-33 mRNA. BCTC (10 μM) significantly abolished menthol-induced up-regulation of TRPM8 mRNA and protein and IL-25 and TSLP mRNA (P < 0.01). TRPM8 protein levels were higher in the supernatants of induced sputum from asthmatic subjects (n = 107) than in those from healthy controls (n = 19) (P < 0.001), and IL-25, TSLP and IL-33 mRNA levels were concomitantly increased (P < 0.001). Additionally, TRPM8 mRNA levels correlated strongly with those of IL-25 and TSLP (P < 0.001), and TRPM8 protein levels were significantly higher in bronchodilator-responsive asthmatic subjects than in nonresponders. Conclusions: TRPM8 may be involved in the airway epithelial cell innate immune response and a molecular target for the treatment of asthma.

OMC-2010 추출물이 마우스의 비장세포 cytokine 생성에 미치는 영향

배기상 ( Gi Sang Bae ),박경철 ( Kyoung Chel Park ),최선복 ( Sun Bok Choi ),조일주 ( Il Joo Jo ),서상완 ( Sang Wan Seo ),김종진 ( Jong Jin Kim ),신용국 ( Yong Kook Shin ),김민선 ( Min Sun Kim ),박규환 ( Kyu Hwan Park ),김현식 ( Hyu 대한본초학회 2012 大韓本草學會誌 Vol.27 No.5

Objective : This study was performed to estimate the effects of OMC-2010 extract on cytokine production in mouse spleen cells. Methods : Mouse spleen cells were pre-treated with ethanol and water extract of OMC-2010 for 1 h, then stimulated with lipopolysaccharide (LPS, 1 μg/ml) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokines. Results : OMC-2010 ethanol extract significantly inhibited the LPS-induced interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and IL-5 mRNA expressions, but not shown such changes in IL-6, IL-4, IL-13. OMC-2010 water extract significantly inhibited the LPS-induced TNF-alpha, and IL-5 mRNA expressions, but not shown such changes in IL-1beta, IL-6, IL-4, IL-13. Conclusions : Theses results could suggest that both ethanol and water OMC-2010 extract could inhibit the TNF-alpha and IL-5 mRNA expression.

정신분열병과 22번 염색체 인터루킨-2 수용체 β-chain 유전자의 연관성

김용구,이민수,김 인,곽동일,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.3

연구배경 : 정신분열병이 유전적이라고 제시하는 많은 역학 연구와 유전자 연구에도 불구하고, 이 질환의 유전방식과 질병유전자는 밝혀져 있지 않다. 본 연구에서는 정신분열병과 22번 염색체 장완의 11.2-12부위에 위치한 Interleukin-2수용체 β chain 유전자간에 유전적 연합을 조사하고자 정신분열병 환자 93명과 정상대조군 97명 대상으로 중합효소연쇄반응을 이용하여 Interleukin-2 수용체 β chain (IL-2Rβ) 유전자의 다형성 분포를 조사하였다. 연구방법 : 환자군은 DSM-Ⅲ-R 진단기준에 따라 임상아형(망상형, 붕괴형, 미붕괴형, 잔류형)으로 분류하였다. 음성 및 양성 정신분열병으로 분류하기위해 Positive and Negative Syndrome Scale(PANSS)을 사용하였다. Genomic DNA를 전혈 임파구에서 추출한 후, IL-2Rβ 유전자좌를 분석하기 위해 dinucleotide(GT)n 염기배열순서를 중합효소연쇄반응을 이용하여 증폭시켰다. 연구결과 : IL-1Rβ의 대립유전자는 모두 8가지 종류이고, guanine-thymine의 반복된 149 염기쌍을 시작으로 151, 153, 155, 157, 159, 161, 163 염기쌍의 형태를 보였다. 정신분열병 환자군과 정상대조군간에 도형접합체 및 이형접합체 빈도의 유의한 차이는 없었다. 환자군과 정상대조군의 대립유전자 분포의 빈도는 통계적으로 유의한 차이가 없었다. 더욱이 각각의 대립유전자 분포에서도 양군간 유의한 차이는 없었다. 또한 동질의 아형으로 분류해 보기위해 임상아형, 양성 및 음성증상군, 가족력의 유무에 따라 비교적 동질적인 표현형을 가진 집단으로 나눈 후 대립유전자 분포를 비교해 보았으나 통계적으로 유의한 차이를 보이지 않았다. 결 론 : 본 연구에서는 Interleukin-2 수용체 β chain 유전자가 정신분열병의 병인론에 관련된다는 가설을 지지할 만한 긍정적 소견을 얻지 못했다. Background : While a significant genetic predisposition to schizophrenia has been proposed, the mode of inheritance or nature of etiological factors is unknown. Previous reports of a genome-wide survey for schizophrenia susceptibility genes have indicated a possible region of linkage on chromosome 22. In order to test the possibility that the interleukin-2 receptor β chain(IL-2Rβ) gene on chromosome 22 is of etiological importance in schizophrenia, a case-control association study was conducted. Methods : Subjects were ninety-three schizophrenic patients with a diagnosis of schizophrenia by DSM-Ⅲ-R criteria and ninety-seven normal controls, Schizophrenic patients were divided by clinical phenotypes such as DSM-Ⅲ-R diagnostic subtypes, positive and negative symptoms, and family history so as to increase the homogeneity of schizophrenics. Genomic DNA was extracted from whole blood lymphocytes according to standard procedures. The DNA was used to study a dinucleotide repeat in the IL-2Rβ gene. To reveal the dinucleotide polymorphism. genomic DNA of subjects was amplified by polymerase chain reactions(PCR). Results : At the IL-2Rβ gene locus, all the previously reported alleles(eight different alleles) of a dinucleotide polymorphism were identified. There was no significant difference between number of heterozygosity in schizophrenic patients and in normal controls. There was no significant difference in the distribution of frequencies of alleles between schizophrenics and normal controls. In addition, there was no significant difference in the allele frequencies among subtypes of schizophrenic patients according to DSM-Ⅲ-R diagnostic subtypes, positive and negative symptoms, and family history. Conclusion : The present study did not detect a difference in frequencies of alleles of a dinucleotide polymorphism at the IL-2Rβ gene locus between schizophrenic patients and normal controls. These results do not supports an evidence that IL-2Rβ gene plays, a major role in the etiology of schizophrenia.

Decreased Pattern-Recognition Receptor-Mediated Cytokine mRNA Expression in Obese Children With Otitis Media With Effusion

Kim, Youn Jung,Cha, Sung Ho,Lee, Ho Yun,Lee, Sun Kyu,Chung, Hee Yong,Yeo, Joon Hyung,Kim, Young Il,Yeo, Seung Geun Korean Society of Otorhinolaryngology-Head and Nec 2014 Clinical and Experimental Otorhinolaryngology Vol.7 No.1

<P><B>Objectives</B></P><P>To assess innate and humoral immune responses in middle ear effusion of obese pediatric patients with otitis media with effusion (OME).</P><P><B>Methods</B></P><P>We evaluated 219 children with OME, of whom 21 were obese and 198 were non-obese. We compared the expression in middle ear effusion of mRNAs encoding toll-like receptors (TLR) 2, 4, 5, and 9; nucleotide-binding oligomerization domains (NOD) 1 and 2; retinoic acid-inducible gene (RIG)-I; interleukins (IL)-6, -10, and -12; interferon (IFN)-γ; and tumor necrosis factor (TNF)-α mRNAs. We also compared the expression of immunoglobulins IgG, IgA, and IgM and the bacterial detection rate in the two groups.</P><P><B>Results</B></P><P>TLR2-mediated expression of IL-6 mRNA, TLR4-mediated expression of IL-6 and IL-10 mRNA, TLR5-mediated expression of IL-6, IL-10, and TNF-α mRNA, TLR9-mediated expression of IL-6 mRNA, and NOD2-mediated expression of IL-6, IL-12, and TNF-α mRNA were significantly lower in obese than in non-obese children (<I>P</I><0.05). However, concentrations of IgG, IgA, and IgM in middle ear effusion were lower in obese than in non-obese children, but none of these differences was significant (<I>P</I>>0.05).</P><P><B>Conclusion</B></P><P>Mean body mass index was higher and pattern-recognition receptor-mediated cytokine mRNA expression was lower in obese than in non-obese children with OME.</P>

A genome-wide by PM₁₀ interaction study identifies novel loci for lung function near <i>BICD1</i> and <i>IL1RN-IL1F10</i> genes in Korean adults

Kim, Hyun-Jin,Seo, Yong-Seok,Sung, Joohon,Chae, Jeesoo,Yun, Jae Moon,Kwon, Hyuktae,Cho, Belong,Kim, Jong-Il,Park, Jin-Ho Elsevier 2020 CHEMOSPHERE - Vol.245 No.-

<P><B>Abstract</B></P> <P>Although several genome-wide interaction studies (GWIS) have been performed in specific European populations to understand the missing link between genetic and environmental factors for lung function, GWIS of Asian samples remain rare. Therefore, we performed a GWIS of exposure to air pollution to identify loci for lung function in Korean adult men. A total of 1826 adult men recruited from two health check-up centers were included in the analysis and the annual mean concentrations of ambient particulate matter with an aerodynamic diameter ≤10 μm (PM<SUB>10</SUB>) were used. In case of forced vital capacity (FVC), one SNP (rs12312730) that passed our genome-wide threshold of <I>p</I>int < 1 × 10–5 was detected in the intronic region of the <I>BICD1</I> gene on chromosome 12. In addition, we found two variants (rs6743376 and rs17042888) located near the <I>IL1RN-IL1F10</I> gene that were involved in the inflammatory response and associated with decreased FVC via interaction with PM<SUB>10</SUB> exposure. A stratified association analysis according to these SNP genotypes showed that PM<SUB>10</SUB> concentrations in subjects with one or two of the risk alleles, compared with those with the non-risk allele, were significantly correlated with a reduction in FVC. This pattern was replicated in another 892 Korean adult samples. The current study reports the first GWIS discovery in an Asian population: the <I>BICD1</I> and <I>IL1RN-IL1F10</I> genes may contribute to the decrease in FVC levels by interacting with PM<SUB>10</SUB> exposure.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Significant interactions between <I>BICD1 or IL1RN-IL1F10</I> and PM<SUB>10</SUB> for FVC were found. </LI> <LI> The several SNPs in these genes were more susceptible to FVC decline by PM<SUB>10</SUB>. </LI> <LI> For FVC, these interaction effects were reproducible in another sample. </LI> </UL> </P>

Adiponectin is a negative regulator of NK cell cytotoxicity.

Kim, Kun-Yong,Kim, Jae Kwang,Han, Seung Hyun,Lim, Jong-Seok,Kim, Keun Il,Cho, Dae Ho,Lee, Myeong-Sok,Lee, Jeong-Hyung,Yoon, Do-Young,Yoon, Suk Ran,Chung, Jin Woong,Choi, Inpyo,Kim, Eunjoon,Yang, Young American Association of Immunologists 2006 Journal of Immunology Vol.176 No.10

<P>NK cells are a key component of innate immune systems, and their activity is regulated by cytokines and hormones. Adiponectin, which is secreted from white adipose tissues, plays important roles in various diseases, including hypertension, cardiovascular diseases, inflammatory disorders, and cancer. In this study the effect of adiponectin on NK cell activity was investigated. Adiponectin was found to suppress the IL-2-enhanced cytotoxic activity of NK cells without affecting basal NK cell cytotoxicity and to inhibit IL-2-induced NF-kappaB activation via activation of the AMP-activated protein kinase, indicating that it suppresses IL-2-enhanced NK cell cytotoxicity through the AMP-activated protein kinase-mediated inhibition of NF-kappaB activation. IFN-gamma enhances NK cell cytotoxicity by causing an increase in the levels of expression of TRAIL and Fas ligand. The production of IFN-gamma, one of the NF-kappaB target genes in NK cells, was also found to be suppressed by adiponectin, accompanied by the subsequent down-regulation of IFN-gamma-inducible TRAIL and Fas ligand expression. These results clearly demonstrate that adiponectin is a potent negative regulator of IL-2-induced NK cell activation and thus may act as an in vivo regulator of anti-inflammatory functions.</P>

In vitro combinatorial anti-proliferative and immunosuppressive effects of <i>Brucea javanica</i> extract with CX-4945 and imatinib in human T-cell acute lymphoblastic leukemia cells

Jung, Jung-Il,Kim, Se Young,Park, Kyeong-Yong,Sydara, Kongmany,Lee, Sang Woo,Kim, Soon Ae,Kim, Jiyeon Elsevier 2018 BIOMEDICINE AND PHARMACOTHERAPY Vol.106 No.-

<P><B>Abstract</B></P> <P>Since 1970, the isolated and identified components of <I>Brucea javanica</I> (L.) Merr. have been known to contain anticancer effects, particularly antileukemic effect. In this study, the inhibitory effect of <I>Brucea javanica</I> (BJ) on cell growth and inflammation was confirmed in human T-cell acute lymphocytic leukemia (T-ALL) cells, and its efficacy as an antileukemic agent was verified. Our results showed that BJ extract induced caspase-dependent apoptosis of T-ALL Jurkat cells through inhibition of the CK2-mediated signaling pathway, while exerting no significant cytotoxicity in normal peripheral blood mononuclear cells. Moreover, BJ extract suppressed the NF-κB signaling pathway, thus, inhibiting the interleukin (IL)-2 expression induced by phorbol 12-myristate 13-acetate (PMA) and phytohemagglutinin (PHA). Notably, combined treatment with BJ extract plus CX-4945 or imatinib exerted enhanced inhibitory effects on T-ALL cell growth and IL-2 production. Overall, these results suggest that BJ extract can be a potent therapeutic herbal agent for T-ALL treatment and prevention of IL-2 mediated inflammatory immune responses.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Brucea javanica</I> extract induces caspase-dependent apoptosis of T-ALL cells. </LI> <LI> <I>Brucea javanica</I> extract the PMA/PHA-mediated NF-κB signaling pathway and IL-2 production. </LI> <LI> <I>Brucea javanica</I> extract suppresses expression of CK2 subunits and Akt phosphorylation. </LI> <LI> Combined treatment exerts enhanced inhibitory effects on T-ALL cell viability and IL-2 production. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

$CD4^+$ Jurkat T 세포주에서 Th1과 Th2 사이토가인 조절에 미치는 황금 유래 Baicalin, Baicalein 및 Wogonin의 효과

김용준,이정치,김홍용,설광화,윤용갑,장선일,Kim Young Jun,Lee Jeong Chi,Kim Hong Yong,Xie Guanghua,Yun Yong Gab,Jang Seon Il 대한동의생리학회 2005 동의생리병리학회지 Vol.19 No.3

In the present study, baicalin, baicalein, and wogonin, a major flavone isolated from Scutellaria Radix were examined for their effects on PMA-induced Interlukin-6 (IL-6), $interferon-\gamma(IFN-\gamma)$, tumor necrosis factor $(TNF)-\alpha$, IL-4, IL-10, and IL-13 productions in the PMA-stimulated $CD4^+$ Jurkat T cells. These three compounds inhibited PMA-induced Th1 cytokine $(IL-6,\;IFN-\gamma,\;TNF-\alpha)$ and Th2 cytokine (IL-4 and IL-13) productions in a concentration-dependent manner. But wogonin, but not baicalin baicalein, increased PMA-induced IL-10 production. These results suggest that baicalin, baicalein, and wogonin, a major flavone modulate Th1 and Th2 cytokine productions in $CD4^+$ Jurkat T cells and these properties may contribute to the anti-atopic dermatitis activity of Scutellaria Radix.