健常人與紅斑性狼瘡病人多形核嗜中性白血球 與單核性細胞間交互作用之研究

Studies on the interactions of polymorphonuclear neutrophils and mononuclear cells in patients with systemic lupus erythematosus and normal

李克仁(Ko-Jen Li)

指導教授 ： 余家利

國立臺灣大學/醫學院/臨床醫學研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.02854

摘要

多形核嗜中性白血球傳統上被認為是主要的吞噬細胞之一，為對抗細菌性病原體的第一道防線。近年來，越來越多的研究報告顯示多形核嗜中性白血球與單核性細胞之間有密切的交互作用存在。但是其詳細的分子機轉及對免疫反應的影響至今仍不清楚。在先前的實驗中，我們以健常人周邊血由來的的單核性細胞為研究對象，發現自體的多形核嗜中性白血球對其Th1 (以IL-2及IFN-γ為代表) 及Th2 (以IL-4及IL-10為代表) 細胞激素的表現均有抑制作用。我們進一步以共軛焦顯微鏡觀察，發現多形核嗜中性白血球與單核性細胞之間有明顯的細胞膜分子的交換現象。在本研究中，我們希望進一步證實此一交換之存在，並以蛋白質體學之研究方法找出可能參與之細胞膜蛋白分子，最後針對此分子在紅斑性狼瘡病人上是否有表現異常加以研究。
研究結果顯示，在流式細胞儀偵測下，多形核嗜中性白血球與單核性細胞之間確實有細胞膜的交換。另外，以西方墨點轉漬法與蛋白質序列分析，我們進一步發現乳鐵蛋白可能為多形核嗜中性白血球與單核性細胞之間交換的分子之ㄧ。我們也發現乳鐵蛋白會抑制健常人或是紅斑性狼瘡病人單核性細胞Th1 (以IFN-γ為代表)細胞激素分泌，但會促進Th2 (以IL-10為代表)細胞激素分泌。相較於在健常人的多形核嗜中性白血球對單核性細胞Th1/Th2細胞激素分泌之影響與乳鐵蛋白類似，在紅斑性狼瘡病人多形核嗜中性白血球對單核性細胞Th2細胞激素分泌有著相反的抑制效果。另外，以流式細胞儀測定發現紅斑性狼瘡病人多形核嗜中性白血球膜上的乳鐵蛋白表現量明顯比健常人少，這些結果顯示紅斑性狼瘡病人的多形核嗜中性白血球上的功能障礙可以導致自體單核性細胞免疫反應的異常。

關鍵字

多形核嗜中性白血球 ； 細胞膜交換 ； 紅斑性狼瘡

並列摘要

Polymorphonuclear neutrophils (PMN) are regarded traditionally as the first line cell component of the body defense mechanisms against bacterial pathogens. Recently, more and more investigations demonstrated that PMN have closely interaction with mononuclear cells (MNC) but its mechanism and contribution in immune response are not clearly understood yet. In previous study, we found autologous PMN exhibited suppression effect on MNC Th1/Th2 cytokine production and significant membrane transfer was observed by confocal microscope. In this study, we want to further confirm membrane transfer in PMN and MNC. Besides, we try to find the protein(s) on PMN involving this transfer and possible defects in SLE.
As the results, we demonstrated the membrane transfer between PMN and MNC by flow cytometry. Furthermore, we used western blot and protein sequencing to find one of the cognate antigens binding with MNC membrane protein on PMN surface, lactoferrin (LF). We also demonstrated that LF can suppress MNC IFN-γ and enhanceIL-10 production which is similar to PMN effect on MNC. In SLE, PMN express less surface LF than normal control and suppress both IFN-γ and IL-10. These results indicate that defects in SLE PMN function lead to aberrant immune response in autologous MNC.

並列關鍵字

PMN ； membrane transfer ； SLE

參考文獻

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