Mycobacterial Heparin-binding Hemagglutinin Antigen Activates Inflammatory Responses through PI3-K/Akt, NF-κB, and MAPK Pathways

김기혜,김화중,조은경,양철수,신아름,전소라,박정규 대한면역학회 2011 Immune Network Vol.11 No.2

Background: Mycobacterium tuberculosis (Mtb) heparin binding hemagglutinin (HBHA) is an Ag known to evoke effective host immune responses during tuberculosis infection. However, the molecular basis of the host immune response to HBHA has not been fully characterized. In this study, we examined the molecular mechanisms by which HBHA can induce the expression of proinflammatory cytokines in macrophages. Methods: HBHA-induced mRNA and protein levels of proinflammatory cytokines were determined in bone marrow-derived macrophages (BMDMs) using RT-PCR and ELISA analysis. The roles of intracellular signaling pathways for NF-κB, PI3-K/Akt, and MAPKs were investigated in macrophage proinflammatory responses after stimulation with HBHA. Results: HBHA robustly activated the expression of mRNA and protein of both TNF-α and IL-6, and induced phosphorylation of NF-κB, Akt, and MAPKs in BMDMs. Both TNF-α and IL-6 production by HBHA was regulated by the NF-κB, PI3-K, and MAPK pathways. Furthermore, PI3-K activity was required for the HBHA-induced activation of ERK1/2 and p38 MAPK, but not JNK, pathways. Conclusion: These data suggest that mycobacterial HBHA significantly induces proinflammatory responses through crosstalk between the PI3-K and MAPK pathways in macrophages.

직업 및 교육수준에 따른 사망원인별 비례사망비의 연도별 추이: 1993-2004년 우리나라 사망등록자료의 분석

김기혜,이경학,이상민,이승연,이예승,임경리,장지은,조상원,최은혜,정성태,진은정,손미아,Kim, Ki-Hye,Lee, Kyung-Hak,Lee, Sang-Min,Lee, Seung-Yeon,Lee, Ye-Seung,Lim, Kyoung-Ree,Chang, Jee-Eun,Cho, Sang-Won,Choi, Eun-Hye,Chung, Sung-Tae,Jin, E 대한예방의학회 2007 예방의학회지 Vol.40 No.1

Objectives : This study investigated the relationship of occupational class and educational background with proportional mortality ratios in Korea. Methods : Mortality was investigated using the entire registered death data from 1993 to 2004, obtained from the Korean National Statistics Office. Proportional mortality ratios (PMRs) for specific diseases were calculated according to the occupational class and educational background of men aged 20-64. Results : Manual workers were found to have higher PMRs for liver disease and traffic accidents, as did the lower educated group. Especially, this study showed trends of an increasing of the wide gap between lower and higher socioeconomic stati for liver disease, traffic accidents, diabetes mellitus and cerebral vascular disease. The mortality for cerebrovascular disease, diabetes mellitus, heart disease, traffic accident and liver disease showed increasing trends according to the calendar year for the lower than the higher social class. Conclusions : The specific conditions that had higher PMRs in the Korean lower social class were liver disease and traffic accidents. Especially, there was an increasing trend for a widening of the gap between manual and non-manual groups in relation to mortality from liver disease, diabetes mellitus and traffic accidents.

Characterization of Proinflammatory Responses and Innate Signaling Activation in Macrophages Infected with Mycobacterium scrofulaceum

김기혜,김태성,Joy G. Lee,박정규,양미소,김진만,조은경,육재민 대한면역학회 2014 Immune Network Vol.14 No.6

Mycobacterium scrofulaceum is an environmental andslow-growing atypical mycobacterium. Emerging evidencesuggests that M. scrofulaceum infection is associated withcervical lymphadenitis in children and pulmonary or systemicinfections in immunocompromised adults. However, the natureof host innate immune responses to M. scrofulaceum remainsunclear. In this study, we examined the innate immuneresponses in murine bone marrow-derived macrophages(BMDMs) infected with different M. scrofulaceum strains includingATCC type strains and two clinically isolated strains(rough and smooth types). All three strains resulted in theproduction of proinflammatory cytokines in BMDMs mediatedthrough toll-like receptor-2 and the adaptor MyD88. Activation of MAPKs (extracellular signal-regulated kinase1/2, and p38, and c-Jun N-terminal kinase) and nuclear receptor(NF)-κB together with intracellular reactive oxygenspecies generation were required for the expression of proinflammatorycytokines in BMDMs. In addition, the rough morphotypesof M. scrofulaceum clinical strains induced higherlevels of proinflammatory cytokines, MAPK and NF-κB activation,and ROS production than other strains. When micewere infected with different M. scrofulaceum strains, thoseinfected with the rough strain showed the greatest hepatosplenomegaly,granulomatous lesions, and immune cell infiltrationin the lungs. Notably, the bacterial load was higherin mice infected with rough colonies than in mice infectedwith ATCC or smooth strains. Collectively, these data indicatethat rough M. scrofulaceum induces higher inflammatoryresponses and virulence than ATCC or smoothstrains

유럽어 사용자를 위한 원격 한국어 코스웨어 개발도구

김기혜(KIM Kihye),윤애선(YOON Aesun) 한국정보과학회 언어공학연구회 1997 한국정보과학회 언어공학연구회 학술발표 논문집 Vol.1997 No.10

르네 마그리트 회화에 있어서의 `역설` 연구

김기혜(Ki-Hye Kim) 현대미술사학회 1993 현대미술사연구 Vol.3 No.-

유치원 종일제 프로그램의 바람직한 운영방안

홍용희,김기혜,강경아,김수정 한국유아교육학회 2007 유아교육연구 Vol.27 No.5

바이오매스 자원으로서의 Lemna Paucicostata의 인공배양조건과 이화학적 특성에 관한 연구

곽정훈,이진의,김기혜,엄혜영,신종서,라창식,Kwag, Jung-Hoon,Lee, Jin-Eui,Kim, Ki-Hye,Eum, Hye-Yeong,Shin, Jong-Suh,Ra, Chang-Six 한국축산환경학회 2010 축산시설환경학회지 Vol.16 No.2

본 연구에서는 Lemna P.의 인공배양 조건을 파악하고 생산된 바이오매스의 영양적 가치에 관하여 조사하였다. Lemna P.의 배양을 위해 총용적 $2,630\;cm^3$, 유효용적 $2,240\;cm^3$의 생물배양조를 이용하였으며 인공광원 6,250 lux, 평균온도 $28^{\circ}C$ 조건에서 인공폐수를 이용하여 배양하였다. 물 유동여부가 Lemna P.의 생장에 미치는 영향을 분석한 결과 하부층의 물 유동이 없는 경우 일일 1.1 이상의 성장률을 보인반면 하부층 물 유동이 있는 경우에는 단지 $0.15\;d^{-1}$의 생장률을 보였다. 또한 인공 폐수내 $NH_4$-N 농도변화와 광 조사시간이 Lemna P. 성장에 마치는 영향을 분석한 결과 광주기 $16h\;d^{-1}$에서의 생장률이 8h 및 $24h\;d^{-1}$에서 보다 높았으며 Lemna P.의 생장은 초기 $NH_4$-N의 농도에 크게 영향을 받는 것으로 나타났다. 초기 $NH_4$-N의 농도가 낮을수록 Lemna P.의 생장률은 높았으나 조단백질 함량은 초기$NH_4$-N 농도에 비례하였다. $NH_4$-N의 농도 2, 10, 50, $100\;mg\;L^{-1}$에서 배양한 후의 Lemna P.의 조단백질 함량은 각각 18, 24, 37, 43%로 50과 $100\;mg\;L^{-1}$ 농도에서 생장한 Lemna P. 바이오매스의 조단백질함량은 현재 단백질원으로 이용되고 있는 아마박(조단백질 함량 35%), 면실박(38%), 대두박(45%)과 비슷한 수준으로 나타났다. 광주기 $16h\;d^{-1}$ 초기 $NH_4$-N 농도 $2\;mg\;L^{-1}$ 이하에서 매우 높은 생장률로 증식한 Lemna P.의 조지방, 조단백질, 조섬유, 조회분, NDF 및 ADF 함량이 각각 2.77, 18.03, 27.02, 20.01, 41.00, 65.68%로 밝혀짐에 따라 우수한 단백질 또는 섬유질 대체자원의 대량 생산이 가능함을 알 수 있었다. 특히 Lemna P.는 식물성 지방보다 동물성 지방에서 검출되는 단일 및 다중 불포화 지방산들을 함유하고 있으며, 가능성 지방산으로 알려진 C18:1n9c, C18:2n6c, C20:5n3 및 C22:2 들을 지나고 있어 가축의 가능성 사료자원으로 활용가치가 높을 것으로 판단되었다. Conditions for artificial culture of Lemna Paucicostata and its nutritional values were examined in this study. Lemna P. was cultured using artificial wastewater and a bioreactor (total volume $2,630\;cm^3$, working volume $2,240\;cm^3$) was operated at conditions of 6,250 lux and $28^{\circ}C$. Water flow affected the growth of Lemna P.: growth rate was very high (more than $1.1\;d^{-1}$) at a condition of no-water movement, but it was very low (less than $0.15\;d^{-1}$) when water moved slowly. The growth of Lemna P. was higher in $16h\;d^{-1}$ light cycle than in Sand $24h\;d^{-1}$, and it was also severely affected by the initial $NH_4$-N levels of wastewater. The growth rate of Lemna P. was high in lower $NH_4$-N level, indicating that the growth rate is in inverse proportion to $NH_4$-N concentration in wastewater. However, the contents of crude protein (CP) of Lemna P. were proportional to the initial $NH_4$-N concentration. The CP contents of Lemna P. cultured at 2, 10, 50 and 100 $NH_4$-N mg $L^{-1}$ was 18, 24, 37, 43%, respectively, showing the Lemna P. cultured at 50 and $100\;mg\;L^{-1}$ had similar protein contents to linseed (CP 35%), cottonseed (CP 38%) and soybean (CP 45%). Fat, protein, fiber, NDF and ADF contents of Lemna P. harvested at conditions of $16h\;d^{-1}$ light cycle and less than $2\;mg\;L^{-1}$ of $NH_4$-N level was 2.8, 18, 27, 20, 41 and 65.7%, respectively. Since the growth rate of Lemna P. was very high (more than $1.1\;d^{-1}$) at those conditions, it was convinced that mass production of valuable protein and fiber sources are feasible. In particular, since the Lemna P. has unsaturated fatty acids found mainly in animal fat as well as beneficial fatty acids to health such as C18:ln9c, C18:2n6c, C20:5n3 and C22:2, the Lemna P. biomass would be a highly valuable alternative feed source to grains.

Respiratory Syncytial Virus Fusion Protein-encoding DNA Vaccine Is Less Effective in Conferring Protection against Inflammatory Disease than a Virus-like Particle Platform

권영만,황혜숙,이영태,김기혜,이유리,김민철,이유나,Fu Shi Quan,Martin L. Moore,강상무 대한면역학회 2019 Immune Network Vol.19 No.3

Formalin-inactivated respiratory syncytial virus (RSV) vaccination causes vaccine-enhanced disease (VED) after RSV infection. It is considered that vaccine platforms enabling endogenous synthesis of RSV immunogens would induce favorable immune responses than non-replicating subunit vaccines in avoiding VED. Here, we investigated the immunogenicity, protection, and disease in mice after vaccination with RSV fusion protein (F) encoding plasmid DNA (F-DNA) or virus-like particles presenting RSV F (F-VLP). F-DNA vaccination induced CD8 T cells and RSV neutralizing Abs, whereas F-VLP elicited higher levels of IgG2a isotype and neutralizing Abs, and germinal center B cells, contributing to protection by controlling lung viral loads after RSV challenge. However, mice that were immunized with F-DNA displayed weight loss and pulmonary histopathology, and induced F specific CD8 T cell responses and recruitment of monocytes and plasmacytoid dendritic cells into the lungs. These innate immune parameters, RSV disease, and pulmonary histopathology were lower in mice that were immunized with F-VLP after challenge. This study provides important insight into developing effective and safe RSV vaccines.

고려홍삼으로부터 분리한 compound K 함유분획에 의한 대식세포의 toll-like receptor-의존성 신호전달로 활성조절 분석

양철수,고성룡,조병구,이지연,김기혜,신동민,육재민,손현수,김영숙,위재준,도재호,조은경,Yang, Chul-Su,Ko, Sung-Ryong,Cho, Byung-Goo,Lee, Ji-Yeon,Kim, Ki-Hye,Shin, Dong-Min,Yuk, Jae-Min,Sohn, Hyun-Joo,Kim, Young-Sook,Wee, Jae-Joon,Do, Jae-Ho 고려인삼학회 2007 Journal of Ginseng Research Vol.31 No.4

본 연구에서는 고려홍삼으로부터 새로 분리한 CK 함유분획을 이용하여 마우스 대식세포에 대한 선천면역반응 조절에 미치는 영향을 조사하였다. 본 연구에서 사용된 농도의 CK 함유분획에서는 세포독성 효과가 관찰되지 않았으며 CK함유 분획의 전처리에 의하여 그람음성세균의 LPS, 또는 CpG-ODN에 의해 유도되는 NF-${\kappa}B$와 MAPK 활성 및 전염증성 사이토카인, NO의 분비가 TLR4 및 TLR9 특이적으로 억제되었다. 이와 같은 결과는 CK 함유분획이 TLR4을 매개로하는 염증반응뿐만 아니라 TLR9을 통한 염증반응에도 영향을 미치는 것으로 해석된다. 따라서 앞으로 CK 함유 분획에 포함된 개별 사포닌 등 시료 성분에 대한 면밀한 분석, 그리고 이들 개별 물질이 각각의 신호전달 체계에 미치는 영향과 그 기작에 대한 연구가 더욱 필요할 것으로 사료되며 염증억제제로서의 개발 가능성을 탐구하기 위한 생체 내에서의 효능 및 작용기전 분석이 요구된다. Compound K (CK), a protopanaxadiol ginsenoside metabolite, was previously shown to have immunomodulatory effects. In this study, we isolated the CK rich fractions (CKRF) from Korean Red Ginseng and investigated the regulation of CKRF-mediated inflammatory signaling during Toll-like receptor (TLR)-mediated cellular activation. Among various TLR ligands, CKRF considerably abrogated TLR4- or TLR9-induced inflammatory signaling. Both LPS and CpG-containing oligodeoxynucleotides (CpG-ODN) stimulation rapidly activates mitogen-activated protein kinases [MAPKs; extracellular signal-regulated kinases 1/2 and p38], NF-${\kappa}B$, and expression of pro-inflammatory cytokines tumor necrosis factor-${\alpha}$, and interleukin-6 in murine bone marrow-derived macrophages (BMDMs) in a time- and dose-dependent manner. Of interest, pre-treatment of CKRF in either LPS/TLR4- or CpG-ODN/TLR9-stimulated macrophages substantially attenuated the LPS-induced inflammatory cytokine production and mRNA expressions, as well as MAPK and NF-${\kappa}B$ activation. To our knowledge, this is the first description of the inhibitory roles for CKRF in TLR4- or TLR9-associated signaling in BMDMs. Collectively, these results demonstrate that CKRF specifically modulates distinct TLR4 and TLR9-mediated inflammatory responses, and further studies are urgently needed for their in vivo roles for potential therapeutic uses, such as in systemic inflammatory syndromes.

Natural Killer and CD8 T Cells Contribute to Protection by Formalin Inactivated Respiratory Syncytial Virus Vaccination under a CD4-Deficient Condition

고은주,이유리,이영태,황혜숙,박윤서,김기혜,강상무 대한면역학회 2020 Immune Network Vol.20 No.6

Respiratory syncytial virus (RSV) causes severe pulmonary disease in infants, young children, and the elderly. Formalin inactivated RSV (FI-RSV) vaccine trials failed due to vaccine enhanced respiratory disease, but the underlying immune mechanisms remain not fully understood. In this study, we have used wild type C57BL/6 and CD4 knockout (CD4KO) mouse models to better understand the roles of the CD4 T cells and cellular mechanisms responsible for enhanced respiratory disease after FI-RSV vaccination and RSV infection. Less eosinophil infiltration and lower pro-inflammatory cytokine production were observed in FI-RSV vaccinated CD4KO mice after RSV infection compared to FI-RSV vaccinated C57BL/6 mice. NK cells and cytokine-producing CD8 T cells were recruited at high levels in the airways of CD4KO mice, correlating with reduced respiratory disease. Depletion studies provided evidence that virus control was primarily mediated by NK cells whereas CD8 T cells contributed to IFN-γ production and less eosinophilic lung inflammation. This study demonstrated the differential roles of effector CD4 and CD8 T cells as well as NK cells, in networking with other inflammatory infiltrates in RSV disease in immune competent and CD4-deficient condition.