論文名稱:

細胞補體調節蛋白於單純皰疹病毒顆粒之結合作用

研究生:

林儀芳  Yi-Fang Lim

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

洪善鈴  Shan-Limg Hung

        學位類別:

碩士

        學校名稱:

國立陽明大學

系所名稱:

口腔生物研究所

            學號:

38917006

          學年度:

90

          語文別:

中文

          出版年:

91

關鍵字:

單純皰疹病毒 

 

細胞補體調節蛋白 

全文說明:

電子全文

        論文頁數:

77

摘要:

補體(complement)是先天免疫反應系統(innate immunity system)部份,補體調節蛋白(complement control protein)可以保護宿主本身的細胞免受補體非專一性的攻擊,有些病毒自感染細胞釋放出來時,病毒外套膜上嵌有宿主的補體調節蛋白,可能藉此保護病毒本身對抗補體的攻擊,如人類免疫不全病毒(human immunodeficiency virus)、巨細胞病毒(cytomegalovirus)和牛痘病毒(vaccinia virus)等都具有這樣的機制。本研究乃探討單純皰疹病毒(herpes simplex virusHSV)第一型(HSV-1)及第二型(HSV-2)是否含有宿主補體調節蛋白。實驗中利用人類子宮頸癌上皮細胞(HeLa)純化出病毒HSV-1-HHSV-2-H,並利用非洲綠猴腎細胞(Vero)純化出病毒HSV-1-VHSV-2-V。利用西方墨澤染色(Western blotting)研究被上述四種純化病毒感染之HeLaVero細胞以及四種純化病毒之病毒蛋白gBgC、宿主細胞蛋白actinCD81補體調節蛋白(CD46CD55CD59)表現情形。HeLa細胞受到HSV-1-HHSV-1-VHSV-2-HHSV-2-V感染後,補體調節蛋白CD46CD59的含量有下降的情形,受到HSV-1-HHSV-1-VHSV-2-V感染後CD55含量下降,而受到HSV-2-H感染後CD55含量不變。純化的HSV-1-HHSV-2-H病毒顆粒上可測到人類補體調節蛋白CD46CD55CD59的存在。Vero細胞不表現CD46CD55CD59HSV-1-VHSV-2-V病毒也測不到這三種蛋白質。HeLaVero細胞和四種純化病毒均可測到宿主細胞之CD81actin。病毒蛋白gBgC在模擬感染(mock-infected)HeLaVero細胞上沒測到,gBgC在被HSV-1-HHSV-1-VHSV-2-HHSV-2-V感染24小時的細胞中的含量比感染8小時的細胞中多,四種純化病毒均可測到gBgC。本研究結果顯示單純皰疹病毒顆粒上,會取得來自宿主細胞之補體調節蛋白CD46CD55CD59,可能可以藉此保護病毒顆粒免於補體系統破壞。

   
 

Complement is part of the innate immune system, and non-specific attacking on host cells is prevented by complement control proteins. Some viruses incorporate host complement control proteins on viral envelopes when viruses are released from infected cell. This incorporation may contribute complement resistance to virus. Human immunodeficiency virus, cytomegalovirus and vaccinia virus have this mechanism. The purpose of this study was to determine the possible incorporation of cellular complement control proteins by herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). HSV-1-H and HSV-2-H were purified from HeLa cells (human cervix epithelioid carcinoma cells); whereas HSV-1-V and HSV-2-V were purified from Vero cells (African green monkey kidney cells). The expression of viral protein gB, gC, cellular protein actin, CD81 and complement control proteins (CD46, CD55 and CD59) in HeLa and Vero cells infected by four purified viruses and these purified virions were analyzed by western blotting. Expression of complement control protein CD46 and CD59 in HeLa cells infected by HSV-1-H, HSV-1-V, HSV-2-H or HSV-2-V was reduced. Expression of CD55 was reduced in HSV-1-H-, HSV-1-V- or HSV-2-V-infected HeLa cells, but was constant in HSV-2-H-infected cells. Human complement control proteins, CD46, CD55 and CD59 were detected in purified HSV-1-H and HSV-2-H virions. CD46, CD55 and CD59 were not detected in Vero cells, HSV-1-V and HSV-2-V virions. Cellular protein CD81 and actin were detected in HeLa cells, Vero cells and four purified virions. Viral proteins, gB and gC, were not detected in mock-infected HeLa or Vero cells. The expression of gB and gC in HSV-1-H-, HSV-1-V-, HSV-2-H- or HSV-2-V-infected cells was higher after infection for 24 hours than for 8 hours. Viral gB and gC were detected in four purified viruses. The results demonstrated that HSV virions acquired host complement control proteins, CD46, CD55 and CD59. This incorporation may provide protection of virions from complement-mediated immune response.