Immune defense mechanism in the central nervous system : The role of Microglia and astrocytes

Matsumoto, Yoh 한림대학교 한림과학원 부설 환경ㆍ생명과학연구소 1995 국제학술회의 Vol.1995 No.-

Pertussis toxin-induced hyperacute autoimmune encephalomyelitis in Lewis rats is correlated with increased expression of inducible nitric oxide synthase and tumor necrosis factor alpha

Ahn, Meejung,Kang, Jongchul,Lee, Yongduk,Riu, Keyzung,Kim, Yong-sik,Jee, Youngheun,Matsumoto, Yoh,Shin, Taekyun 제주대학교 방사능이용연구소 2001 연구보고 Vol.15 No.-

자기면역성 뇌척수염(experimental autoimmune encephalomyelitis, EAE)은 뇌조직항원을 면역한 후 야기되는 염증성 질병으로 사람 다발성결화증의 한 모델로 연구되고 있다. 자기면역성 뇌염의 시작은 뇌조직항원에 반응하는 림프구가 중추신경계에 침윤되면서 마비를 나타내는데 이 과정 중에는 여러 종류의 pro-inflammatory mediator (tumor necrosis factor-alpha (TNF-α)와 inducible nitric oxide synthase (iNOS)등)가 관여하는 것으로 알려지고 있다. 이 연구에서는 염증의 진행 단계에 따라 염증 유도 또는 염증 억제의 상반된 기능을 갖는 것으로 알려진 TNF-α와 iNOS가 심급성 뇌척수염 진행에 어떠한 영향을 미치는지를 조사하였다. 뇌염을 유도하기 위한 항원으로는 랫트 척수 조직 유제를 complete Freund adjuvant와 혼합하여 뒷 발바닥에 주사하였으며 심한 뇌척수염을 유도하기 위하여 pertussis toxin(500ng/ea)을 면역하는 날 복강내로 주사하고 매일 체중과 마비 정도를 확인하였다. 독소를 주사한 실험군에서는 대조군(11일)에 비해 마비의 시작이 빨랐으며(9일), 대조군은 자연 회복하는 반면 독소룰 주사한 실험군에서는 모두 폐사하였다. 척수 조직 내 TNF-α 와 iNOS의 양적인 변화를 조사하기 위하여 Competitive PCR과 Western blot를 이용하였으며, 세포형을 구분하기 위하여 면역염색을 이용하였다. Competitive PCR결과 TNF-α는 PT를 투여한 자기면역성뇌척수염의 심한 마비기(EAE,G3)에서 PT를 투여하지 않은 대조군보다 약 5배가 증가하였으며(p<0.01), Western blot결과 iNOS는 PT를 투여한 군에서 정상조직에 비해 약 6배가 증가하였고, PT를 투여하지 않은 군에 비해서는 약 3개바 증가하였다(p<0.01). 면역염색결과 PT를 투여하지 않은 랫트보다 투여한 랫트의 척수조직에서 iNOS 양성 세포가 약 15배가 증가하였으며(p<0.01), 또한 연속절편에서 이들 세포가 큰포식세포임을 확인하였다. 이상의 결과를 종합해 볼 때, 자기면역성 뇌척수염의 초기 유도과정에서는 TNF-α와 iNOS는 염증의 약화에 관여됨을 알 수 있었다. The involvement of inducible nitric oxide synthase (iNOS) and tumor necrosis factor alpha (TNF-α), which have diverse roles in the progression of autoimmune disease models, was studied in pertussis toxin (PT-induced hyperacute experimental autoimmune encephalomyelitis (EAE) in Lewis rats. The expression of TNF-α mRNA(increased 5 fold, p<0.01) and iNOS protein (3 fold, p<0.01) was much greater in the spinal cords with PT(+) EAE at the peak stage of EAE than in those with PT(-) EAE, as shown by competitive PCR and Western blot analysis, respectively. Immunohistochemistry showed that the majority of EDI-positive macrophages in EAE lesions contained iNOS, and that three were many more iNOS-positive cells in the CNS lesions of PT(+) rats than in those of PT(-) rats. These findings suggest that PT-induced hyperacute EAE is partly mediated by the enhanced expression of iNOS and TNF-α in the early stages of rat EAE.

뇌염모델에서 Protein Kinase C의 발현에 관한 연구

김형민,신태균,Matsumoto, Yoh,Tanuma, Naoyuki 제주대학교 방사능이용연구소 1997 연구보고 Vol.11 No.-

We examined the cellular distribution and regulation of protein kinase C delta in the spinal cord of rats with experimental autoimmune encephalomyelitis(EAE). Northern blotting showed that levels of PKC delta mRNA was significantly increased at the peak stage of EAE and its expression decreased to the level of non-immunized conrols at the recovery stage. By in situ hybridization, signals of PKC delta was localized on the inflammatory cells including mainly T cells and macrophages as well as some brain cells. This finding suggests a hypothesis that increased expression of PKC on inflammatory cells in EAE lesions is associated with the regulation of signaling pathway involving T cell activation at the early stage of EAE and/or apoptosis on inflammatory cells including T cells at recovery stage, which lead to spontaneous recovery.

Enhanced expression of constitutive and inducible forms of nitric oxide synthase in autoimmune encephalomyelitis

Tae Kyun Shin,Yoh Matsumoto,Naoyuki Tanuma,Hyung Min Kim,Myung Bok Wie,Chang Jong Moon,Seung Joon Kim 대한수의학회 2000 Journal of Veterinary Science Vol.1 No.1

Mechanism of experimental autoimmune encephalomyelitis in Lewis rats

Taekyun Shin,Meejung Ahn,Yoh Matsumoto 대한해부학회 2012 Anatomy & Cell Biology Vol.45 No.3

Experimental autoimmune encephalomyelitis (EAE) in Lewis rats is an acute monophasic paralytic central nervous system disease, in which most rats spontaneously recover from paralysis. EAE in Lewis rats is induced by encephalitogenic antigens, including myelin basic protein. EAE is mediated by CD4⁺ Th1 cells, which secrete pro-inflammatory mediators, and spontaneous recovery is mediated by regulatory T cells. Recently, it was established that classically activated macrophages (M1 phenotype) play an important role in the initiation of EAE, while alternatively activated macrophages (M2 phenotype) contribute to spontaneous recovery from rat EAE. This review will summarize the neuroimmunological aspects of active monophasic EAE, which manifests as neuroinflammation followed by neuroimmunomodulation and/or neuroprotection, with a focus on the role of alternatively activated macrophages.

Alternatively activated M2 macrophages increase in early stages of experimental autoimmune myocarditis in Lewis rats

Oh, Hanseul,Ahn, Meejung,Matsumoto, Yoh,Shin, Taekyun The Korean Society of Veterinary Science 2013 大韓獸醫學會誌 Vol.53 No.4

To better understand the role of macrophages in early stages of experimental autoimmune myocarditis (EAM), we compared the expression of inducible nitric oxide synthase (iNOS) and arginase-1, markers for classically activated M1 and alternatively activated M2 macrophages, respectively, in the hearts of EAM-affected and control rats. Immunohistochemical evidence revealed that both iNOS-positive and arginase 1-positive macrophages were found in EAM lesions, while some cells were co-localized with both markers. This finding suggests that the increased level of arginase-1, which is partly from M2 macrophages, contributes to the modulation of EAM, possibly through the reduction of nitric oxide in the lesion.

Enhanced expression of three forms of nitric oxide synthase in autoimmune central nervous system disease

Kim, Seungjoon,Moon, Changjong,Wie, Myungbok,Kim, Hyungmin,Tanuma, Naoyuki,Matsumoto, Yoh,Shin, Taekyun 濟州大學校 農科大學 動物科學硏究所 2000 動物科學論叢 Vol.15 No.1

To elucidate the role of nitric oxide synthase (NOS) in the pathogenesis of experimental autoimmune encephalomyelitis (EM), we analyzed the expression of constitutive neuronal (nSOS), endothelial (eNOS) and inducible NOS (iNOS), an enzyme of NO production, in the spinal cords of rats with EAE. We further examined the structural interaction between apoptotic cells and spinal cord cells including neurons and astrocytes, which are potent cell types of NO production in the brain. Western blot analysis shows that three isoforms of NOS significantly increased in the spinal cords of rats at the peak stage of EAE, while minimal amounts of these same enzymes are identified in normal rat spinal cords. Immunohistochemical study showed that brain cells includmg neurons and astrocytes revealed an increased expression of either nSOS and/or eNOS during the peak and recovery stages of EAE, while iNOS was found mainly in the inflammatory macrophages in the perivascular EAE lesions. Double labeling showed that apoptotic cells had intimate contacts with either neurons or astrocvtes, which are major cell types to express nNOS and eNOS constitutively. Our data suggests that both constitutive nSOS and eNOS as well as iNOS, possibly producing NO, plays an important role in the recovery of EAE.

Increased expression of P53 and Bax in the spinal cords of rats with experimental autoimmune encephalomylitis

문창종,김승준,위명복,김희석,정종태,박전홍,지영흔,Tanumab, Naoyuki,Matsumoto, Yoh,신태균 濟州大學校 農科大學 動物科學硏究所 2000 動物科學論叢 Vol.15 No.1

The expression of pro-apoptotic molecules, p53 and Bax, in the spinal cord of rats with experimental autoimmune encephalomyelitis (EAE) was examined. Apoptosis was confirmed by the terminal deoxynucleotidyl transferasemediated dUTP nick end-labeling (TUNEL) method. TUSEL (+) apoptotic cells were mainly either ED1 (+) macrophages or T-cells in the parenchyma of E N . Western blot analysis showed that both p53 and Bax expression significantly ( p < 0.01) increased in the spinal cords of EAE rats at the peak stage, and thereafter declined. An immunohistochemical study showed that inflammatory cells (notably T cells) in the parenchyma express p53 and Bax, while brain cells, includng neurons and glia, were devoid of these nuclear staining of these molecules. The nuclear expression of p53 largely matches apoptotic cells in the parenchyma of EAE. These finchngs suggest that pro-apoptotic molecules, p53 and Bax, may play an important role in eliminating T cells in the parenchyma in EAE.

뇌염모델에서 Protein Kinase C의 발현에 관한 연구

Shin, Tae-Kyun,Kim, Hyung-Min,Tanuma, Naoyuki,Matsumoto, Yoh 한국수의병리학회 1997 한국수의병리학회지 Vol.1 No.1

Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.