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Expression Profiles of Molecules Involved in Antigen Presentation in Gangliosides-stimulated Primary Astrocytes from Rat brain
DC Field | Value | Language |
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dc.contributor.author | 윤, 희정 | - |
dc.date.accessioned | 2011-04-14T01:53:29Z | - |
dc.date.available | 2011-04-14T01:53:29Z | - |
dc.date.issued | 2005 | - |
dc.identifier.uri | http://repository.ajou.ac.kr/handle/201003/2287 | - |
dc.description.abstract | Astrocytes can function as antigen-presenting cells (APC) by expressing class II major histocompatibility complex (MHC) antigens and co-stimulatory molecules upon activation. In vitro, astrocytes are activated by lipopolysaccharides (LPS), and interferon-gamma (IFN-gamma). We previously reported that gangliosides, membrane components of brain cells, especially neurons, can activate brain glial cells in vitro, thus could play roles in brain physiology and pathology. In this study, I tested whether gangliosides could induce the expression of immune molecules related to antigen presentation, using cultured rat brain glial cells, astrocytes and microglia. I treated cells with 50 ug/ml gangliosides mixture (Gmix) for 6 h to 3 d, and assayed the mRNA expression of class II transactivator (CIITA) and class II major histocompatibility (MHCII) molecules, using RT-PCR and real-time PCR. Gangliosides induce CIITA mRNA expression, followed by MHC II in astrocytes, but not in microglia. Critical for the development of mature APC capability is surface expression of class II MHC molecules and co-stimulatory molecules expression. Cell surface molecules of central importance to costimulation are CD40, ICAM-1, CD80 (B7-1) and CD86 (B7-2). In astrocytes, mRNA expressions of the above molecules were constitutive and that of CD86 was further induced by treatment of Gmix. Those expression patterns are the same in IFN-gamma-treated cells. Next, I tested the surface expression of MHC II by FACS and immunocytochemistry analysis. Gmix treatment failed to induce it in both FACS and immunocytochemistry analysis, in contrast to IFN-gamma treatment. This imply that Gmix could induce the mRNA expressions of CIITA and MHCII, but they could not induce surface expression of MHCII. To clarify the mechanisms, I assayed the molecules involved in it, including invariant chain (Ii) and the non-classical class II molecule RT1-DMs. Treatment of Gmix could not induce the mRNA expression of Ii, while IFN-gamma could. Still further studies are needed, I presumably suggest that Gmix are defective in fully presenting antigens due to a lack of expressing Ii molecules. | - |
dc.description.abstract | 신경교세포는 lipopolysaccharides(LPS)와 interferon-γ(IFN-γ) 자극에 의한 class II major histocompatibility complex (MHCII)와 동시 자극 물질(co-stimulatory molecules)의 활성화에 의해 항원 제시 세포(APC)로 작용할 수 있다. 최근 보고에 의하면 뇌세포의 막성분인 gangliosides(Gmix)에 신경교세포를 자극하여 활성화 시키나 이미 다른 자극제에 의해 활성화된 MHCII와 동시 자극 물질의 발현을 저해한다. 그러나 Gmix가 쥐 뇌의 신경교세포로부터 면역 관련 물질들을 발현시키는데 단독으로는 어떤 작용을 하는지에 대해서는 보고가 되어 있지 않다. 본 연구에서 Primary microglia와 astrocytes를 Gmix로 자극하면 CIITA와 MHCII는 astrocytes에서는 발현되나 microglia에서는 발현되지 않음을 RT-PCR로 확인하였으며, astrocytse에서 MHCII의 발현도는 Real-time PCR로 관찰하였다. 그러나 astrocytes에서 발현된 MHCII는 IFN-g와 LPS에 의해 발현된 MHCII와 달리 세포 표면으로 발현되지 않는 것을 FACS와 immunohistochemistry 실험에서 확인하였다. IFN-g와 Gmix에 의한 동시 자극 물질 발현은 같은 경향을 보이는 것을 확인하였다. 그러나 MHCII가 세포 표면으로 발현되는 경로에 관여하는 invariant chain(Ii)과 RT1-DMs을 RTPCR로 관찰하였을 때, RT1-DMs의 발현은 IFN-g와 Gmix 모두 같은 경향을 보였으나, Ii는 Gmix에 의해서 발현되지 않았다. 지금까지 결과를 모두 종합하여, Gmix는 이전에 보고된 바와 같이 신경교세포를 자극하여 활성화시키나 신경교세포가 APC로 작용하기 위한 MHCII의 발현에 있어, mRNA은 증가시키나 항원 제시 경로에서 Ii의 발현이 일어나지 않아 APC로 작용하지 못하게 된다는 결론을 얻을 수 있었다. | - |
dc.description.tableofcontents | "TABLE OF CONTENTS
ABSTRACT = ⅰ TABLE OF CONTENTS = ⅲ LIST OF FIGURES = ⅴ LIST OF TABLE = ⅶ LIST OF ABBREVIATION = ⅷ Ⅰ. INTRODUCTION = 1 Ⅱ. MATERIAL AND METHOD = 5 A. Reagents = 5 B. Cell cultures = 5 C. Flow cytometry (FACS) = 6 D. RNA isolation and Reverse Transcription Polymerase Chain Reaction (RT-PCR) = 6 E. SYBR Green Real-time PCR = 8 F. Immunocytochemistry = 9 Ⅲ. RESULTS = 11 A. Gmix induces the mRNA of CIITA and MHCII in primary rat astrocytes but not in primary rat microglia. = 11 B. Gmix failed to induce MHCII expression at cell surface. = 16 C. B7-2 transcript was induced, while those of B7-1, CD40 and ICAM-1 were constitutively expressed, not being further induced by Gmix. = 19 D. Individual components of Gmix have the same effect on the transcript and surface expression of antigen-presenting molecules. = 22 E. Gmix failed to induce Ii transcript in astrocytes. = 25 Ⅳ. DISCUSSION = 28 Ⅴ. CONCLUSION = 33 REFERENCES = 34 국문요약 = 41" | - |
dc.language.iso | en | - |
dc.title | Expression Profiles of Molecules Involved in Antigen Presentation in Gangliosides-stimulated Primary Astrocytes from Rat brain | - |
dc.title.alternative | Ganglioside 자극 신경 성상세포 활성에서 항원 제시 물질 발현에 관한 연구 | - |
dc.type | Thesis | - |
dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000000285 | - |
dc.subject.keyword | Ganglioside | - |
dc.subject.keyword | Astrocyte | - |
dc.subject.keyword | antigen-presentig cell | - |
dc.subject.keyword | MHCII | - |
dc.subject.keyword | CIITA | - |
dc.subject.keyword | co-stimulatory molecules | - |
dc.subject.keyword | Gmix | - |
dc.subject.keyword | Ii | - |
dc.description.degree | Master | - |
dc.contributor.department | 대학원 의학과 | - |
dc.contributor.affiliatedAuthor | 윤, 희정 | - |
dc.date.awarded | 2005 | - |
dc.type.local | Theses | - |
dc.citation.date | 2005 | - |
dc.embargo.liftdate | 9999-12-31 | - |
dc.embargo.terms | 9999-12-31 | - |
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