자궁내막증 병변에서의 Cytokine mRNA의 발현 양상

이택후(Taek Hoo Lee),김광수(Gwang Soo Kim),김일규(Il Gyu Kim),전상식(Sang Sik Chun),조영래(Young Lae Cho) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.9

목적: 자궁내막증은 최근에 그 빈도가 급격하게 증가되고 있으나 아직까지 병인과 치료법이 확실하게 정립되지 못하고 있는 부인과 내분비의 대표적인 질환이다. 최근 들어서 자궁내막증 환자의 불임의 원인으로 cytokine이 밀접하게 관여됨이 밝혀지고 있고 자궁내막증의 병인으로 복강내로 역류된 월경혈에 의한 복강내의 국소적 염증에 대한 개체의 반응정도와 면역체계의 변화가 중요하게 인정되고 있으나 아직까지 정확한 기전은 밝혀져 있지 않기 때문에 이에 저자는 자궁내막증 환자의 복수 내에서의 cytokine의 정량측정과 골반내 자궁내막증 병변조직에서의 cytokine mRNA의 발현 양상을 조사하여 자궁내막증 환자의 병인분석을 시도하였다. 연구방법: 자궁내막증 진단을 받고 복강경 혹은 개복 수술을 받은 30명의 환자에서 총 60개의 자궁내막증 병변이라고 의심되는 조직을 채취하여 이를 RT-PCR법을 이용하여 Cytokine 유전자의 발현양상을 조사하였다. 또한 7례의 정상대조군과 23례의 자궁내막증 환자의 복수를 ELISA법을 이용하여 정량분석을 시도하였다. 결과: 자궁내막증 환자의 복수내에서 IL-6와 IL-10의 농도는 자궁내막증의 임상적 중증도에 따라서 의미있게 증가되어 있었으나 IFN-γ, TNF-α, IL-1β, 그리고 IL-5의 농도는 정상 대조군에 비해서 변화가 없었다. 모든 예의 심부 자궁내막증 병변조직과 표재성 자궁내막증 병변조직에서 IL-1β cytokine mRNA의 발현을 볼 수 있었다. IL-5와 IL-6은 표재성 병변 12개 중에서 각각 2개의 black lesion에서만 발현을 볼 수 있었으며 IL-10은 표재성 병변에서는 12개중 2개에서 그리고 심부성 병변에서는 8개중 1개의 조직에서만 발현되었다. IFN-γ는 표재성 병변에서는 전혀 발현이 없었으나 심부성 병변에서는 8개중 4개의 조직에서 발현이 되었으며 TNF-α는 표재성 병변에서는 red 및 black 병변에서 각각 1개의 조직에서만 발현이 되었으나 심부성 병변에서는 역시 8개중 4개의 조직에서 발현이 되었다. 결론: 표재성 병변이 골반강내에 착상하여 염증성 반응이 일어날 수 있는 원인이 제공되면 IL-1β 혹은 TNF-α같은 염증성 cytokine이 분비가 되며 이로 인해 생성되는 단핵세포의 chemotactic factor에 의해 대식세포의 증가와 활성화가 이루어지고 이어서 복강내에 IL-6 등의 cytokine이 증가되며 마지막으로 여러 가지 증가된 cytokine에 대한 반대 반응으로 IL-10이 증가됨을 추정할 수가 있겠으며 이러한 가정은 앞으로 cytokine을 이용한 치료적 응용의 기초적인 연구로서 중요한 의미를 제공할 수 있다고 하겠다. Objective: The pathogenesis of endometriosis is generally accepted that retrograde menstruation and alterations in the local pelvic immune environment. This study was performed to help elucidate what kind of role various cytokines might play in the pathogenesis of endometriosis. Method: Concentrations of peritoneal fluid cytokines were compared in 7 women with normal pelvic finding and 23 women with endometriosis by enzyme-linked immunosorbent assay(ELISA). The patterns of cytokine mRNA expression in 8 ovarian endometrioma and 12 superficial pelvic endometriosis lesions were investigated by reverse transcription-polymerase chain reaction(RT-PCR) amplification method. Result: Both IL-6 and IL-10 levels in peritoneal fluid specimens with endometriosis tended to be higher than normal. However, there were no significant differences between peritoneal fluid concentrations of IFN-γ, TNF-α, IL-1β, and IL-5 of women with and without endometriosis. The levels of IL-6 and IL-10 were significantly higher in peritoneal fluid of women with severe endometriosis compared to women with mild endometriosis. IL-1β mRNA was expressed in all of 8 deep and 12 superficial endometriosis lesions. IL-5 and IL-6 mRNA were expressed in only two black lesions respectively, however, both were not expressed in the all deep lesions. Expressions of IL-10 mRNA occurred in one red and one black lesion while this was expressed in only one of the deep lesions. TNF-α mRNA was expressed in one red and one black lesion of 12 superficial lesions compared with four of the deep lesions. There was the difference between kinds of increased cytokines in the peritoneal fluid and those of expressed cytokines in the endometriotic lesions of patients with endometriosis. Conclusion: This study supports the concept that local immunologic factors may be important in the pathogenesis and pathophysiology of endometriosis. The pattern of cytokine mRNA expression of endometriotic lesions would seem to indicate that proinflammatory cytokines such as IL-1β and TNF-α are responsible for the development or progression of endometriosis.

Cytokine-cytokine receptor interactions in the highly pathogenic avian influenza H5N1 virus-infected lungs of genetically disparate Ri chicken lines

부 티 하오,Hong Yeojin,츠엉앵득,Lee Jiae,Lee Sooyeon,송기덕,Cha Jihye,Dang Hoang Vu,Tran Ha Thi Thanh,Lillehoj,Hyun S.,홍영호 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.3

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry as well as the economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for HPAIV resistance. Therefore, in this study, we investigated gene expression related to cytokine-cytokine receptor interactions by comparing resistant and susceptible Ri chicken lines for avian influenza virus infection. Methods: Ri chickens of resistant (Mx/A; BF2/B21) and susceptible (Mx/G; BF2/B13) lines were selected by genotyping the Mx dynamin like GTPase (Mx) and major histocompatibility complex class I antigen BF2 genes. These chickens were then infected with influenza A virus subtype H5N1, and their lung tissues were collected for RNA sequencing. Results: In total, 972 differentially expressed genes (DEGs) were observed between resistant and susceptible Ri chickens, according to the gene ontology and Kyoto encyclopedia of genes and genomes pathways. In particular, DEGs associated with cytokine-cytokine receptor interactions were most abundant. The expression levels of cytokines (interleukin-1β [IL-1β], IL-6, IL-8, and IL-18), chemokines (C-C Motif chemokine ligand 4 [CCL4] and CCL17), interferons (IFN-γ), and IFN-stimulated genes (Mx1, CCL19, 2’-5’-oligoadenylate synthaselike, and protein kinase R) were higher in H5N1-resistant chickens than in H5N1-susceptible chickens. Conclusion: Resistant chickens show stronger immune responses and antiviral activity (cytokines, chemokines, and IFN-stimulated genes) than those of susceptible chickens against HPAIV infection. Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry as well as the economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for HPAIV resistance. Therefore, in this study, we investigated gene expression related to cytokine-cytokine receptor interactions by comparing resistant and susceptible Ri chicken lines for avian influenza virus infection.Methods: Ri chickens of resistant (Mx/A; BF2/B21) and susceptible (Mx/G; BF2/B13) lines were selected by genotyping the Mx dynamin like GTPase (Mx) and major histocompatibility complex class I antigen BF2 genes. These chickens were then infected with influenza A virus subtype H5N1, and their lung tissues were collected for RNA sequencing.Results: In total, 972 differentially expressed genes (DEGs) were observed between resistant and susceptible Ri chickens, according to the gene ontology and Kyoto encyclopedia of genes and genomes pathways. In particular, DEGs associated with cytokine-cytokine receptor interactions were most abundant. The expression levels of cytokines (interleukin-1β [IL-1β], IL-6, IL-8, and IL-18), chemokines (C-C Motif chemokine ligand 4 [CCL4] and CCL17), interferons (IFN-γ), and IFN-stimulated genes (Mx1, CCL19, 2’-5’-oligoadenylate synthaselike, and protein kinase R) were higher in H5N1-resistant chickens than in H5N1-susceptible chickens.Conclusion: Resistant chickens show stronger immune responses and antiviral activity (cytokines, chemokines, and IFN-stimulated genes) than those of susceptible chickens against HPAIV infection.

왼온엉덩동맥 허혈 후 재관류 시간경과에 따른 생쥐의 혈청 내 친염증성 cytokine의 변화

박경신(Kyung shin Park),김성재(Sung Jae Kim),서윤경(Younkyoung Seo) 대한체질인류학회 2016 해부·생물인류학 (Anat Biol Anthropol) Vol.29 No.1

허혈 재관류 손상은 허혈이 이루어진 후 재관류되면서 일어난다. 허혈 재관류 손상의 주요 원인은 활성산소기와 활성화된 면역세포가 분비하는 cytokine이다. Cytokine은 감염이나 면역반응, 염증 등의 반응을 조절하는 세포 사이의 신호전달물질로, 염증반응을 증가시키는 것을 친염증성 cytokine이라 한다. 본 실험에서 저자들은 허혈후 재관류 경과에 따른 친염증성 cytokine인 IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNγ의 변화를 알아보았다. 12주령의 수컷 생쥐를 이용하여 정상대조군과 2시간 허혈군, 4시간 허혈군, 6시간 허혈군으로 구분하였고, 총 96마리의 생쥐를 사용하였다. 허혈처치는 왼온엉덩동맥을 혈관집게를 이용하여 차단하였고, 각 허혈군은 재관류 경과에 따라 0시간, 2시간, 4시간, 8시간, 16시간으로 재분류하였다. 재관류 시간에 따라 생쥐를 마취시킨 후 심장에서 혈액을 채취하였다. 혈청을 분리한 후 ELISA 분석방법을 이용하여 IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNγ의 농도를 측정하여 다음과 같은 결과를 얻었다. 13개의 cytokine 농도는 허혈시간에 따라 그리고 재관류 시간에 따라 유의한 차이가 있었다. 허혈 2시간군에서 IL-1α와 IL-3는 정상대조군보다 유의성 있게 증가되었고, 재관류 후 MIP-1α를 제외한 모든 cytokine들은 증가하였으며, MCP-1과 TARC은 재관류 16시간에 가장 높은 농도로 관찰되었다. 허혈 4시간군에서 TARC만이 정상대조군보다 유의성 있게 증가되었으며, 재관류 후 모든 cytokine들은 재관류 4시간 이후에 감소하는 경향으로 관찰되었다. 허혈 6시간군에서 IL-2, IL-3, MCP-1, TARC은 정상대조군보다 유의성 있게 증가되었고, 재관류 16시간에 IL-3와 MCP-1이 유의성 있게 증가되었다. 이상의 결과를 종합하면, 허혈은 친염증성 cytokine을 정상대조군보다 증가시키는 것을 알 수 있었고, 특히 2시간과 6시간 허혈군에서 관찰된 IL-1α, IL-3, MCP-1, TARC은 재관류 후기까지 비교적 높은 농도로 유지되는 것을 알 수 있었다. Ischemia-reperfusion injury arises from the restoration of blood supply after ischemia. Both reactive oxygen species and various cytokines produced by activated immune cells are the primary causal risk factors for ischemic injury. Cytokines are intercellular signaling substances for regulating any infection, immune reactions and inflammation, and pro-inflammatory cytokines adversely affect any diseases through an increase in inflammatory reaction. This study was conducted to investigate whether the periods of reperfusion after ischemia result in any changes of pro-inflammatory cytokines in the serum, including IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, Eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNδ. A total of 96 male mice aged at 12 weeks was used in this study, and the groups of ischemia were divided into the following three different groups: 2-hour, 4-hour, and 6-hour ischemia groups. For the object of ischemic injury, the left common iliac artery was clamped by vascular clamp, each ischemia group was subdivided into 5 different groups according to the periods of reperfusion: 0-, 2-, 4-, 8-, and 16-hour reperfusion time. Blood samples after general anesthesia were collected from the mice hearts, and the serum was separated from them. The concentration of pro-inflammatory cytokines (IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, Eotaxin, MCP-1, MDC, MIP-1α, RANTES, TARC, IFNδ) in the serum was measured by ELISA, and the following results were acquired. The concentrations of the 13 pro-inflammatory cytokines were significantly different in accordance with the periods of ischemia and the reperfusion time. In 2-hour ischemia group, IL-1α and IL-3 were increaed compared to normal control group, and 12 cytokines were increased followed by reperfusion except for MIP-1α. MCP- 1 and TARC were expressed as the highest concentration in the 16-hour reperfusion time. In 4-hour ischemia group, TARC was significant differences with normal control group, and the concentration of 13 cytokines were decreased after 4-hour reperfusion time. In 6-hour ischemia group, IL-2, IL-3, MCP-1 and TARC were increased, compared to normal control group, and IL-3 and MCP-1 were increased in 16-hour reperfusion time. To sum up, ischemia increased the pro-inflammatory cytokines compared to normal control group and in the 2-hour and 6-hour ischemia groups, IL-1α, IL-3, MCP-1 and TARC were increased until the late reperfusion time.

인체 대장상피세포에 대한 이질 아메바의 접촉 차단이 친염증성 Cytokine 유전자 발현 유도에 미치는 영향

김정목,정현채,이경미,진영주,김영전,송인성,김정룡 大韓消化器病學會 1997 大韓消化器病學會誌 Vol.30 No.2

HCC : PE-056 ; Change of cytokine profile following transarterial chemotherapy in patients with hepatocellular carcinoma

( Min Ju Kim ),( Jeong Won Jang ),( Jung Hyun Kwon ),( Chan Ran You ),( Nam Ik Han ),( Chang Don Lee ),( Hyun Suk Jung ),( Dong Wook Jekar ),( Seung Ok Lee ),( Kyu Won Chung ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background: Although alterations in cytokine profile after therapy can affect the outcome and prognosis of cancer patients, there are no comprehensive data on multiple cytokine profiles during transarterial chemotherapy (TACE). Methods: Cytometric bead immunoassay was used to simultaneously measure 13 cytokines (interleukin [IL]-12p70, IFN-γ, IL-17A, IL-2, IL-10, IL-9, IL-22, IL-6, IL-13, IL-4, IL-5, IL-1β, and TNF-α) in the sera of 83 patients with hepatocellular carcinoma (HCC) and 18 healthy controls. Those cytokines were serially monitored at baseline, day 3, day 7, and 2 months after TACE in 63 evaluable patients. Results: Serum levels of IL-17A and IL-5 were higher in HCC patients than healthy controls, whereas IL-22 and IL-1β levels were lower in HCC patients. Although the overall patterns of cytokine changes were diverse at each set date, certain cytokines specifically increased after TACE, with the early-phase increase in IL-6 and IL-22 levels and late-phase increase in IL-4, IL-6, and IL-10 levels. With relatively minor changes of cytokines after TACE, Childs B/C group had higher IL-6 and lower IL-22 levels than Childs A group. Patients with larger tumors (>5 cm) had higher IL-6 levels at baseline and showed a transient but significant early-phase increase in IL-6 levels as well as late-phase increase in IL-4, IL-10, and IL-13 levels after TACE. With regard to hepatic events, IL-2 and IL-22 levels at baseline were predictive of grade 3 or more hepatic toxicity, while those levels of IL-6 and IL-13 significantly increased at day 3 in patients suffering from post- TACE hepatic morbidity. Conclusions: TACE induces various changes of multiple cytokines. Distinct panels of cytokine changes are not uniform, influenced by treatment-induced inflammation, underlying liver function, and HCC stage. The increased Th2 cytokine profiles after TACE suggests the immune suppression in patients with large tumor and post-treatment hepatitis.

HCC : PE-056 ; Change of cytokine profile following transarterial chemotherapy in patients with hepatocellular carcinoma

( Min Ju Kim ),( Jeong Won Jang ),( Jung Hyun Kwon ),( Chan Ran You ),( Nam Ik Han ),( Chang Don Lee ),( Hyun Suk Jung ),( Dong Wook Jekarl ),( Seung Ok Lee ),( Kyu Won Chung ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Although alterations in cytokine profile after therapy can affect the outcome and prognosis of cancer patients, there are no comprehensive data on multiple cytokine profiles during transarterial chemotherapy (TACE). Methods: Cytometric bead immunoassay was used to simultaneously measure 13 cytokines (interleukin [IL]-12p70, IFN-γ, IL-17A, IL-2, IL-10, IL-9, IL-22, IL-6, IL-13, IL-4, IL-5, IL-1β, and TNF-α) in the sera of 83 patients with hepatocellular carcinoma (HCC) and 18 healthy controls. Those cytokines were serially monitored at baseline, day 3, day 7, and 2 months after TACE in 63 evaluable patients. Results: Serum levels of IL-17A and IL-5 were higher in HCC patients than healthy controls, whereas IL-22 and IL-1β levels were lower in HCC patients. Although the overall patterns of cytokine changes were diverse at each set date, certain cytokines specifically increased after TACE, with the early-phase increase in IL-6 and IL-22 levels and late-phase increase in IL-4, IL-6, and IL-10 levels. With relatively minor changes of cytokines after TACE, Childs B/C group had higher IL-6 and lower IL-22 levels than Childs A group. Patients with larger tumors (>5 cm) had higher IL-6 levels at baseline and showed a transient but significant early-phase increase in IL-6 levels as well as late-phase increase in IL-4, IL-10, and IL-13 levels after TACE. With regard to hepatic events, IL-2 and IL-22 levels at baseline were predictive of grade 3 or more hepatic toxicity, while those levels of IL-6 and IL-13 significantly increased at day 3 in patients suffering from post- TACE hepatic morbidity. Conclusions: TACE induces various changes of multiple cytokines. Distinct panels of cytokine changes are not uniform, influenced by treatment-induced inflammation, underlying liver function, and HCC stage. The increased Th2 cytokine profiles after TACE suggests the immune suppression in patients with large tumor and post-treatment hepatitis.

PM(10)이 A549 Cells에서 전염증성 Cytokine발현에 미치는 영향

김정호 ( Jung Ho Kim ),전효근 ( Hyo Keun Jeon ),김미경 ( Mi Kyeong Kim ),경선영 ( Sun Yong Kyung ),안창혁 ( Chang Hyeok An ),이상표 ( Sang Pyo Lee ),박정웅 ( Jung Woong Park ),정성환 ( Sung Hwan Jeong ) 대한결핵 및 호흡기학회 2006 Tuberculosis and Respiratory Diseases Vol.60 No.6

연구배경: 미세먼지는 여러 가지 유기물과 무기물의 복합체로 그 구성 성분이 시간과 장소에 따라 다르고 모양과 크기도 일정하지 않으며, 특히 지름 10㎛이하의 미세먼지 (particulate matter 10; PM(10))는 흡입이 가능한 입자의 크기여서 하부기관지 및 폐의 가스-교환부분까지 침착하여 호흡기계에 손상을 일으킬 수 있다. 본 연구에서는 황사에 포함된 PM(10)과 비황사 시기에 포집된 PM(10)이 폐상피세포주에 작용하여 전염증성 사이토카인(proinflammatory cytokine) 및 cytokine messenger RNA(mRNA)의 발현에 어떤 영향이 있는지를 관찰하여 기관지 천식과 만성 폐쇄성 폐질환등 호흡기 질환의 증상 악화기전에 미치는 역할을 규명하고자 하였다. 연구방법: 공기 포집기(HV 500F, sibata model)를 이용하여 황사와 비황사 기간에 하루 6시간씩 실외의 장소에서 대기분진을 membrane filter에 포집한 다음, PM(10)입자를 추출하고 폐암 상피세포주인 A549 cells(한국세포은행주)에 PM(10)을 농도에 맞게(10㎍/㎖, 100㎍/㎖, 500㎍/㎖) 노출시켰다. 각각의 노출된 세포로부터 interleukin(IL)-1α, IL-1β, IL-8, granulocyte macrophage colony stimulating factor(GM-CSF)의 mRNA를 역전사중합효소연쇄반응(reverse transcriptase polymerase chain reaction; RT-PCR) 방법으로 측정하였다. 결과: 황사 및 비황사 기간 중 포집된 PM10을 가했을 시 가하지 않은 대조군에 비하여 IL-1α, IL-1β, IL-8, granulocyte macrophage colony stimulating factor (GM-CSF)의 m`RNA와 cytokine의 발현이 유의하게 높았으며, 황사 기간의 고농도의 PM(10)에 노출된 세포의 IL-1α mRNA는 비황사 기간의 PM(10)에 노출된 세포의 mRNA보다 증가되어 있었다. 결론: PM(10)은 A549 cells에서 전염증성 사이토카인의 발현을 증가시키고 비황사 기간보다 황사 기간 중 대기 중에서 채취한 PM(10)에 노출된 A549 cells에서 일부의 전염증성 사이토카인의 mRNA발현을 더욱 증가시키는 것을 알 수 있었다. 따라서 황사 기간의 PM(10)에 의한 일부의 전염증성 사이토카인의 발현 증가가 만성 호흡기 질환의 증상 악화기전에 연관되어 있을 가능성을 시사하였다. Background: PM(10)(Particulate matter with a diameter < 10㎛), which is characterized by different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also produce unique particulate matter in affected areas. This study investigated the cytokine produced by A549 epithelial cells exposed to particles collected during both the Asian dust pfenomenon and ambient air particles in a non-dusty period. Method: Air samples were collected using a high volume air sampler(Sibata Model HV500F) with an air flow at 500ℓ/min for at least 6 hours. The cytokine messenger RNA(mRNA) was measured using a reverse transcriptase polymerase chain reaction(RT-PCR). The A549 cells were exposed to 10 to 500㎍/㎖ of a suspension containing PM(10) for 24 hours. Each was compared with those in the non-exposed control cells. Result: The mRNA levels of interleukin(IL)-1α, IL-Iβ , IL-8, and the granulocyte macrophage colony stimulating factor(GM-CSF) increased after veing exposed to PM(10) in the ambient air particles, compared with those in the non-exposed control cells. The increase in IL-1α and IL-8 were dose dependent at a PM(10) concentration between 100㎍/㎖ and 500㎍/㎖. The mRNA level of IL-8 in the A549 epithelial cells was higher during the in the Asian dust period(500㎍/㎖) than during the non dust period. Conclusion: A549 cells exposed to the PM(10) collected during the Asian dust period produce more proinflammatory cytokine than during non-dusty period. This cytokine enhances the local inflammatory response in the airways and can also contribute to the systemic component of this inflammatory process. (Tuberc Respir Dis 2006; 60: 663-672)

Lipopolysaccharide가 배양 각질형성세포 및 말초혈액 단핵구의 Cytokines 유전자 발현에 미치는 영향

윤기성,전재복,김도원,정상립,김문규 慶北大學校 醫科大學 1995 慶北醫大誌 Vol.36 No.4

목적 : 라포다당류(lipopolysaccharde LPS)는 glycolipids 복합물로 인체내의 여러 면역세포, 특히 혈액내의 단핵구/대식세포에 강력하고 다양한 형태의 자극제로 작용, 각종 사이토카인(cytokine)을, 포함한 여러 매개물질의 분비를 야기하며, 배양세포의 자극제로도 흔히 이용된다. 이에 LPS로 자극된 배양 각질형성 세포및 말초혈액 단핵구가 어떤 사이토카인 유전자를 발현하며 양자간에 그 차이는 어떤가를 알아보고자 한다. 대상 및 방법 : 포피절제술과 말초혈액 채취를 통해 얻어진 배양 각질형성세포 및 단핵구를 역전사 및 중합효소 연쇄반응을 사용하여 사이토카인 유전자 발현을 측정하였다. 결과 : 말초혈액 단핵구에서는 5㎍/㎖의 LPS로 자극한 경우 6 시간 및 24 시간 후 모두에서 대조군인 GAPDH를 포함하여 실험한 IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α, TGF-α, TGF-β, GM-CSF 등 11 종류의 모든 사이토카인 유전자 발현을 볼 수 있었고 LPS를 주지 않은 군에서는 6 시간 및 24 시간 후 IL-2와 IL-6을 제외한 9 종류의 사이토카인 유전자 발현을 볼 수 있었으며 그 발현의 정도는 약했다. 각질형성세포에서는 LPS 처치 전 24 시간 동안 BPE를 제거한 각질형성세포 배양액을 사용했을 때 LPS를 주지 않은 경우와 5㎍/㎖의 LPS로 자극한 경우 모두 6 시간 및 24 시간 후 다 같이 어떤 유전자 발현도 볼 수 없었으며, 25㎍/㎖의 LPS로 자극한 경우 6 시간 후에는 유전자 발현이 없었으나 24 시간 후에는 IL-1α, IL-1β, IL-8, IL-10, TGF-α, TGF-β 및 GM-CSF의 유전자 발현을 볼 수 있었고, 이들의 발현 정도는 LPS의 농도가 높을수록 강하게 나타났다. 결론 : LPS가 배양 각질형성세포 및 말초혈액 단핵구의 사이토카인 유전자 발현을 자극하나 양세포간에는 뚜렷한 차이가 있었으며, 각질형성세포에서는 BPE가 cytokine 유전자 발현에 상당한 변수로 작용할 수 있음을 관찰할 수 있었다. Lipopolysaccharde(LPS), a complex glycolipids, is the major component of the outermost membrane of Gram-negative bacteria. There is much interest in LPS because it provides a potent and pleiotrophic stimulus for immune cells, both in vitro and in vivo. It has been often used as the stimulant for several cultured cells, and is known to stimulate the keratinocytes to produce several cytokines. To investigate the effects of LPS on cytokine gene expression of cultured keratinocytes and peripheral blood mononuclear cells (PBMC), we examined the transcripts of cytokine genes using the reverse transcription and polymerase chain reaction (RT-PCR) method with 13 cytokine-specifci primers. The results were as follows : 1. LPS in a concentration of 5 ㎍/㎖ increased the transcription of all the cytokines we examined in PBMC, i.e., interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-α, TGF-β and granulocyte/macrophage-colony stimulating factor (GM-CSF). 2. In keratinocytes cultured in the media without bovine pituitary extract(BPE) for the last 24 hours, LPS in a concentration of 5 ㎍/㎖ did not stimulate any cytokine gene expression. However, LPS in a concentration of 25 ㎍/㎖ stimulated cytokine gene expression of IL-1α, IL-1β, TGF-α, TGF-β and GM-CSF-after 24 hours. 3. In keratinocytes cultured in the media containing BPE, LPS in both concentration of 5 ㎍/㎖ and 25 ㎍/㎖ stimulated cytokine gene expression of IL-1α, IL-1β, IL-8, IL-10, TGF-α, TGF-β and GM-CSF after 6 and 24 hours, respectively.

Inflammatory Cytokines and Their Prognostic Ability in Cases of Major Burn Injury

허준,양형태,전욱,김종현,신선희,강희정,김현수 대한진단검사의학회 2015 Annals of Laboratory Medicine Vol.35 No.1

Background: Major burn injuries induce inflammatory responses and changes in the levels of various cytokines. This study was conducted to assess early changes in the serum levels of inflammatory cytokines after burn injury, identify cytokines associated with mortality, and characterize correlations among cytokines. Methods: Blood samples of 67 burn patients were collected on days 1 and 3 after burn injury, and the concentrations of 27 cytokines were measured using the Bio-Plex Suspension Array System (Bio-Rad Laboratories, USA). Blood samples of 25 healthy subjects were used as controls. We analyzed statistical differences in the concentrations of each cytokine between the control and patient groups, between day 1 and day 3, and between survival and nonsurvival groups. Correlations among 27 cytokines were analyzed. Results: Median concentrations of granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin 1 receptor antagonist (IL-1RA), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin 15 (IL-15), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein 1β (MIP-1β), and vascular endothelial growth factor (VEGF) were significantly higher in burn patients than in controls. IL-1RA, IL-6, and MCP-1 levels were significantly higher in the nonsurvival group than in the survival group on day 1 after burn injury. Correlation analysis of 27 cytokines showed different relationships with one another. Stronger correlations among interferon γ (IFN-γ), IL-2, IL-4, IL-7, IL-12p70, and IL-17 were found. Conclusions: IL-1RA, IL-6, and MCP-1 may be used as prognostic indicators of mortality in burn patients and the increase in cytokine concentrations is induced by interactions within a complex network of cytokine-related pathways.

역전사 중합효소연쇄반응을 이용한 태아혈액의 Interleukin-1β, Tumor Necrosis Factor-α, Transforming Growth Factor-β의 발현양상

최석태 ( Seok Tae Choi ),윤보현 ( Bo Hyun Yoon ),신희철 ( Hee Chul Syn ) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.12

Background: For the successful pregnancy, the feto-maternal immunologic bidirectional interaction should be programed to accept the fetal semiallograft and assist the feto-placental growth in early pregnancy. It has been known that these signaling materials for interacting are cytokines. Up to the present, the potential roles of cytokines in normal pregnancy are, first, as a signal material of maternal immunologic recognition of fetal semiallograft, second, as a substance like growth- factor to promote the feto-placental growth and differentiation, third, as a material to alter the maternal immunity for fetal survival, fourth, as a mediator to initiate or prevent the labor. We have proposed the hypothesis that various cytokines may be expressed simultaneously, and then, may regulate the maintenance of pregnancy by interacting with each other harmonically after midpregnancy. Because the maintenance of pregnancy and the mechanism of labor depend on the total effect of various cytokines rather than the effect of one specific cytokine. Objectives: To support a potential role of the fetus itself for participating in the bidirectional cytokine regulatory network, and support our hypothesis that various cytokines may be expressed simultaneously, and then, may regulate the maintenance of pregnancy by interacting with each other harmonically after midpregnancy. Study dasign: For exploring the actual immunologic status of fetus itself in utero, we have chosen the seventeen fetal blood samples which have been taken at the cordocentesis or at the delivery without labor after midpregnancy(20th~39th weeks of gestation). After extracting the RNA from these fetal blood samples, we have tried to demonstrate the expression of interleukin-1β and tumor necrosis factor-a mRNA, and simultaneously, demonstrate the expression of transforming growth factor-β mRNA by using reverse transcriptase-po1ymerase chain reaction(rt-PCR) technique. Results: In this study we found that the expression of interleukin-1β mRNA was demonstrated from as early as 22nd weeks to term gestation, and the expression of tumor necrosis factor-α mRNA was demonstrated from as early as 2300rd weeks to term gestation, and the expression of transforming growth factor-β mRNA was demonstrated from as early as 21st weeks to term gestation. Conclusion: On the basis of these findings, first, we support the result of recent study that the fetus itself may participate in the bidirectional cytokine regulatory network after midpregnancy, second, we prove the possible hypothsis that various cytokines may be expressed simultaneously, and then, may regulate the maintenance of pregnancy by interacting with each other harmonically after mid pregnancy.