論文名稱:

調控第一型單純皰疹病毒感染口腔細胞產生細胞激素所牽涉之機制

 

Mechanisms involved in modulation of cytokine production in oral cells after herpes simplex virus type 1 infection

研究生:

鄧宇晴  Yu-Ching Teng

 

(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-Ling 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:

39017001

          學年度:

91

          語文別:

中文

          出版年:

92

關鍵字:

第一型單純皰疹病毒  herpes simplex virus type 1 (HSV-1)

 

牙齦角化細胞  gingival keratinocyte (GK)

 

牙齦造纖維細胞  gingival fibroblast (GF)

 

細胞激素  cytokine

 

白素-6  interleukin-6 (IL-6)

 

酵素連結免疫吸附定量法  enzyme-linked immunosorbent assay (ELISA)

 

膠體電泳位移分析  electrophoretic mobility shift assay (EMSA)

 

訊息傳遞路徑  signal transduction pathway

全文說明:

(全文限校內瀏覽)

 

電子全文

        論文頁數:

90

摘要:

  第一型單純皰疹病毒(herpes simplex virus type 1HSV-1)通常造成口腔感染,而導致口炎(gingivostomatitis)及唇皰疹(herpes labialis)。細胞激素 (cytokine)為對抗HSV-1感染的一種免疫反應。已知HSV-1可以感染許多種不同類型的細胞,而使細胞分泌不同的細胞激素,然而對受到HSV-1感染的人類口腔細胞分泌細胞激素的研究較缺乏,因此本論文探討人類的牙齦角化細胞(gingival keratinocyteGK)及牙齦造纖維細胞(gingival fibroblastGF)受到HSV-1感染後,細胞分泌白素-6 (interleukin-6IL-6)白素-8 (IL-8)白素-1β (IL-1β)的情形及可能牽涉的機轉。主要利用酵素連結免疫吸附定量法(enzyme-linked immunosorbent assayELISA)偵測細胞分泌細胞激素量的改變。結果發現受到HSV-1感染的GF細胞分泌IL-6的量增加,但GK細胞受到HSV-1感染後不會分泌IL-6。含抗HSV-1抗體的中和血清可以完全抑制HSV-1感染GF細胞所引起IL-6分泌增加的現象,而以紫外線照射或以加熱方式而失去病毒斑形成能力的HSV-1感染GF細胞後仍能增加細胞分泌IL-6的能力,顯示HSV-1不需要完成生活史 (life cycle)即可刺激GF細胞分泌IL-6。實驗結果亦顯示純化的病毒表面蛋白gD即能夠刺激GF細胞分泌IL-6此外,訊息傳遞路徑NF-κB抑制劑pyrollidine dithiocarbamate (PDTC)N-acetyl-L-cysteine (NAC) 會抑制假性感染的GF細胞及受HSV-1感染後的GF細胞分泌IL-6的能力;而以膠體電泳位移分析(electrophoretic mobility shift assayEMSA)方法分析NF-κB (nuclear factor kappa B)活性的變化,證實HSV-1感染會活化NF-κB,使GF細胞核萃取物與含κB siteDNA結合的活性增加,顯示GF細胞受HSV-1感染而增加IL-6的分泌可能是經由NF-κB訊息傳遞路徑。此外,GF細胞受HSV-1感染後會增加IL-8的分泌,但感染後的GK細胞IL-8的分泌降低,而GFGK細胞受HSV-1感染後皆不會分泌IL-1β。最後,干擾素-γ (interferon-γIFN-γ)會加強受HSV-1感染的GF細胞分泌更多的IL-6,而受HSV-1感染的GK細胞同時受到IFN-γ的刺激後仍不會分泌IL-6,但IFN-γ不會影響受感染的GK細胞分泌IL-8的情形。由本論文的研究結果得知受HSV-1感染的GFGK細胞分泌細胞激素IL-6IL-8的情形不同。其中GFHSV-1感染分泌IL-6增加的機制可能牽涉NF-κB訊息傳遞路徑,而病毒蛋白gD即可刺激GF細胞分泌IL-6

 

Herpes simplex virus type 1 (HSV-1) usually causes oral infection, and results in gingivostomatitis and herpes labialis. Cytokine is one of the immune response against HSV-1 infection. HSV-1 induces expression of various cytokines in many different cell types, but little is known regarding cytokines expressed by oral cells after HSV-1 infection. The purposes of this study were to investigate the secretion of interleukin-6 (IL-6), IL-8 and IL-1β in gingival keratinocytes (GK) and gingival fibroblasts (GF) after HSV-1 infection and to examine the possible mechanisms involved. The cytokine production was measured by an enzyme-linked immunosorbent assay (ELISA). The results showed that HSV-1 induced IL-6 secretion in GF, but not in GK. This phenomenon was completely inhibited by serum which contained anti-HSV-1 antibody. The ultraviolet (UV)- or heat-treatment abolished the plaque forming activity of HSV-1. The UV- or heat-inactivated viruses were still able to induce IL-6 secretion in GF. This suggests that complete life cycle was not required to induce IL-6 secretion in GF. Moreover, purified viral gD alone was able to induce IL-6 secretion in GF. Inhibitors, pyrollidine dithiocarbamate (PDTC) and N-acetyl-L-cysteine (NAC), against NF-κB pathway reduced IL-6 secretion of mock-infected and infected GF. HSV-1 infection also enhanced nuclear protein binding to the κB site DNA as analyzed by electrophoretic mobility shift assay (EMSA). The combined results suggest that induction of IL-6 by HSV-1 in GF may be through NF-κB pathway. In addition, HSV-1 induced IL-8 secretion in GF, whereas HSV-1 reduced IL-8 secretion in GK. IL-1β was not detected in GF and GK after HSV-1 infection. Finally, interferon-γ (IFN-γ) synergistically enhanced IL-6 secretion in GF that infected with HSV-1. However, IFN-γ upregulated IL-6 secretion on HSV-infected GF, but IFN-γ was not affect IL-8 secretion on infected GK. In conclusion, secretion of cytokines, IL-6 and IL-8, was different in GF and GK after HSV-1 infection. HSV-1 induced IL-6 secretion in GF through mechanisms involving NF-κB signal transduction pathway. Viral gD may induce IL-6 secretion in GF.

   

       發表著作

Teng Y-C, Chen Y-T, Cheng Y-Y, Hung S-L. Requirements for the upregulation of interleukin-6 by herpes simplex virus-infected gingival fibroblasts. Viral Immunology 2005; 18(1): 170-178.