本文選題：白念珠菌 + 口腔上皮細(xì)胞　； 參考：《北京協(xié)和醫(yī)學(xué)院》2016年博士論文

【摘要】：念珠菌是臨床常見的機(jī)會(huì)性致病真菌,其中白念珠菌是引起人體口腔及陰道等部位黏膜感染的主要念珠菌。眾所周知,口腔及陰道部位的黏膜上皮細(xì)胞是抵抗念珠菌感染的“第一道防線”。前期研究發(fā)現(xiàn),上皮細(xì)胞抵抗白念珠菌感染主要通過激活炎性信號(hào)通路(如ERK/JUK/p38等信號(hào)通路)、釋放炎性細(xì)胞因子等發(fā)揮免疫功能。鑒于臨床上口腔念珠菌病更易發(fā)生在免疫低下的病人、更易造成系統(tǒng)播散感染,但陰道念珠菌病更易發(fā)生在免疫正常女性、少見系統(tǒng)播散感染,故不同部位的上皮細(xì)胞可能存在不同的抗白念珠菌感染的機(jī)制。白念珠菌胞壁成分ALS3及SSA1與黏附功能有關(guān),在誘導(dǎo)上皮細(xì)胞產(chǎn)生抗念珠菌免疫的過程中起到重要的作用。前期已有研究證明敲除ALS3可降低誘導(dǎo)口腔上皮細(xì)胞分泌產(chǎn)生炎性細(xì)胞因子及炎性信號(hào)通路蛋白的表達(dá)。但卻很少有實(shí)驗(yàn)研究ALS3敲除對陰道上皮細(xì)胞誘導(dǎo)產(chǎn)生炎性細(xì)胞因子的影響及可能涉及到的炎性通路。也很少有實(shí)驗(yàn)研究SSAl敲除對上皮細(xì)胞產(chǎn)生炎性細(xì)胞因子的影響及可能涉及到的炎性通路。本實(shí)驗(yàn)主要研究白念珠菌感染口腔上皮細(xì)胞及陰道上皮細(xì)胞誘導(dǎo)炎癥細(xì)胞因子的產(chǎn)生及ERK信號(hào)通路蛋白激活的分子機(jī)制的差異,同時(shí)探討白念珠菌胞壁成分AIs3蛋白、Ssa1蛋白在此免疫過程中存在的可能作用機(jī)制。第一章 口腔和陰道上皮細(xì)胞抗白念珠菌感染的免疫差異目的：探索口腔上皮細(xì)胞及陰道上皮細(xì)胞體外感染白念珠菌免疫反應(yīng)的異同性。方法：口腔上皮細(xì)胞(leuk-1)、陰道上皮細(xì)胞(VK2/E6E7)進(jìn)行體外培養(yǎng),給予白念珠菌(SC5314)不同菌量的感染。觀察不同部位來源的上皮細(xì)胞在非感染及白念珠菌感染狀態(tài)下免疫效應(yīng)分子及通路蛋白表達(dá)的異同,檢測白念珠菌感染不同上皮細(xì)胞時(shí),其細(xì)胞毒性、炎性細(xì)胞因子分泌及p-ERK1/2、p-MKP1蛋白表達(dá)的差異。結(jié)果：口腔和陰道上皮細(xì)胞,在體外培養(yǎng)的形態(tài)上存在很多共性：倒置顯微鏡下見鋪路石樣外觀,多角形,大小不一,形態(tài)不規(guī)則,細(xì)胞核呈圓形或卵圓形。細(xì)胞經(jīng)過瑞氏-吉姆薩復(fù)合染液染色后,都可呈現(xiàn)出較為典型的細(xì)胞質(zhì)粉紅色,細(xì)胞核藍(lán)(黑)色的表現(xiàn)。生長曲線測定結(jié)果,口腔leuk-1細(xì)胞與陰道VK2/E6E7細(xì)胞的生長增殖過程存在差異,Leuk-1細(xì)胞的增殖速度較VK2/E6E7細(xì)胞快；兩組細(xì)胞在生長增殖的過程中細(xì)胞直徑呈現(xiàn)出類似“拋物線”形狀的變化圖；但leuk-1細(xì)胞直徑整體仍較VK2/E6E7細(xì)胞偏大。白念珠菌感染狀態(tài)下,死亡細(xì)胞沿菌絲分布。細(xì)胞損傷實(shí)驗(yàn)結(jié)果顯示,同一種細(xì)胞的損傷度與白念珠菌感染的菌量及感染時(shí)間有關(guān),即損傷度與白念珠菌菌量、時(shí)間成正比。兩組細(xì)胞對比,白念珠菌對不同上皮細(xì)胞的損傷具有特異性：同一菌量感染狀態(tài)下,白念珠菌對陰道上皮細(xì)胞VK2/E6E7的損傷較重,而對口腔上皮細(xì)胞leuk-1的損傷較輕。炎性細(xì)胞因子的檢測顯示：白念珠菌在口腔上皮細(xì)胞與陰道上皮細(xì)胞中均可刺激誘導(dǎo)GM-CSF、G-CSF、IL-1α、IL-1β、RANTES、MIP-3α、IL-8的分泌量增多,但亦存在一定差異,口腔上皮細(xì)胞在未感染狀態(tài)時(shí)可產(chǎn)生多種細(xì)胞因子,在低劑量白念珠菌感染時(shí)即可產(chǎn)生較多的分泌量；而陰道上皮細(xì)胞在未感染時(shí),多數(shù)因子分泌量極少,即使在白念珠菌感染后,多數(shù)細(xì)胞因子的分泌量仍增幅不大。兩種細(xì)胞出現(xiàn)細(xì)胞因子最大誘導(dǎo)量的白念珠菌刺激菌量不同。在口腔感染過程中, IL-8、GM-CSF、MIP-3α及RANTES的分泌高峰出現(xiàn)在白念珠菌感染菌量／細(xì)胞量(MOI)=0.01時(shí),G-CSF、IL-1β的分泌高峰出現(xiàn)在MOI=0.1時(shí),IL-1 α的分泌高峰出現(xiàn)在MOI=1時(shí)。而陰道上皮細(xì)胞在感染過程中,IL-8βGM-CSF的分泌高峰出現(xiàn)在MOI=0.01時(shí),G-CSF的分泌高峰出現(xiàn)在MOI=0.1,而IL-1α、IL-1β及MIP-3α的分泌高峰出現(xiàn)在MOI=1時(shí)。IL-4及IL-12兩種細(xì)胞因子,未顯示明顯的白念珠菌劑量依賴性或是規(guī)律性的趨勢。在白念珠菌感染口腔和陰道上皮細(xì)胞2h后,分別檢測其p-ERK1/2及p-MKP1蛋白的表達(dá)情況,顯示白念SC5314感染組的p-ERK1/2及p-MKP1蛋白的表達(dá)都較對照組高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論：白念珠菌感染口腔和陰道這兩種不同部位上皮細(xì)胞所產(chǎn)生的細(xì)胞損傷及炎性細(xì)胞因子的含量并不相同,上皮細(xì)胞抗白念珠菌的免疫應(yīng)答存在部位差異性。第二章 白念珠菌ALS3敲除株感染口腔和陰道上皮細(xì)胞后免疫學(xué)機(jī)制的比較研究目的：探索白念珠菌ALS3基因在白念珠菌刺激口腔上皮細(xì)胞及陰道上皮細(xì)胞產(chǎn)生抗白念珠菌免疫反應(yīng)的過程中的作用,及其存在的可能的免疫機(jī)制差異。方法：體外分別培養(yǎng)口腔上皮細(xì)胞和陰道上皮細(xì)胞,給予白念珠菌SC5314(WT株)及白念珠菌ALS3敲除株(als3△)不同菌量的感染。測定白念珠菌WT株及als3△株分別感染不同上皮細(xì)胞時(shí),其細(xì)胞毒性、炎性細(xì)胞因子及ERK1/2、MKP1通路蛋白的表達(dá),對比不同菌量、不同菌株、不同來源細(xì)胞的免疫反應(yīng)的差異。結(jié)果：白念珠菌ALS3基因敲除后,其菌絲的生長不受影響,與SC5314野生株比較,無統(tǒng)計(jì)學(xué)差異(p0.05)。但對比WT及als3△感染上皮細(xì)胞的結(jié)果,als3△對細(xì)胞的損傷明顯降低(p0.05),且其細(xì)胞損傷程度呈劑量依賴型,即MOI值越大,損傷也越大。als3△感染口腔上皮細(xì)胞后,GM-CSF、G-CSF、,IL-1α、IL-1β、MIP-3α和IL-8細(xì)胞因子的產(chǎn)生量均較WT感染組降低(p0.05)。als3△感染陰道上皮細(xì)胞后,除了RANTES及MIP-3 a外的其他細(xì)胞因子,都較WT組降低(p0.05)。對比分析WT株分別感染兩種不同來源上皮細(xì)胞誘導(dǎo)產(chǎn)生G-CSF、RANTES、MIP-3 α、IL-8的結(jié)果,發(fā)現(xiàn)口腔上皮細(xì)胞leuk-1組這些細(xì)胞因子的產(chǎn)量高于陰道細(xì)胞VK2/E6E7組(p0.05)；而als3△感染時(shí),兩組細(xì)胞間上述細(xì)胞因子的產(chǎn)量并無統(tǒng)計(jì)學(xué)差異(p0.05)�？谇簧掀ぜ�(xì)胞分別感染白念珠菌WT與als3△ 2h后,WT感染組與als3△感染組中p-ERK1/2蛋白的表達(dá)結(jié)果無統(tǒng)計(jì)學(xué)差異(p0.05);但WT感染組中p-MKP1蛋白的表達(dá)量高于als3△感染組的表達(dá)量,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。在陰道上皮細(xì)胞VK2/E6E7分別感染白念珠菌WT與als3△的蛋白檢測中,WT感染組p-ERK1/2蛋白及p-MKP1蛋白的表達(dá)量均較als3△感染組的表達(dá)量高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論：白念珠菌ALS3基因的表達(dá)可以誘導(dǎo)口腔上皮細(xì)胞和陰道上皮細(xì)胞產(chǎn)生多種細(xì)胞因子。且als3△刺激兩組細(xì)胞產(chǎn)生某些炎性細(xì)胞因子的能力有一定的差異。在白念珠菌感染口腔上皮細(xì)胞誘導(dǎo)免疫反應(yīng)的過程中,A1s3蛋白不完全通過激活ERK免疫信號(hào)通路增強(qiáng)p-MKP1蛋白的表達(dá)來參與免疫反應(yīng),而在白念珠菌感染陰道上皮細(xì)胞的過程中,Als3蛋白通過激活ERK通路誘導(dǎo)p-MKP1蛋白的表達(dá)增強(qiáng)來參與免疫反應(yīng)。表明口腔上皮細(xì)胞及陰道上皮細(xì)胞在應(yīng)對白念珠菌Als3蛋白誘導(dǎo)的免疫反應(yīng)中,存在部位性差異。第三章 白念珠菌SSA1敲除株感染口腔和陰道上皮細(xì)胞后免疫學(xué)機(jī)制的比較研究目的：探討白念珠菌SSAl基因表達(dá)在刺激口腔上皮細(xì)胞及陰道上皮細(xì)胞產(chǎn)生抗白念珠菌免疫效應(yīng)過程中的作用。方法：體外培養(yǎng)口腔、陰道上皮細(xì)胞,分別用白念珠菌SC5314(WT)及白念珠菌SSAl基因敲除株(ssa1△)進(jìn)行刺激。測定不同菌量刺激兩種不同上皮細(xì)胞時(shí)在細(xì)胞損傷、炎性細(xì)胞因子分泌、通路蛋白p-ERK1/2及p-MKP1表達(dá)的變化。對比不同組別間的差異,分析可能存在的機(jī)制。結(jié)果：體外培養(yǎng)白念珠菌SSAl敲除株,2h后測量菌絲長度較WT株明顯縮短(p0.05)。ssa1△在感染上皮細(xì)胞24h后無法形成菌落,只有散在的菌絲,經(jīng)臺(tái)盼藍(lán)進(jìn)行細(xì)胞染色后,可見死亡細(xì)胞沿菌絲分布。對比WT感染上皮細(xì)胞的結(jié)果,ssa1△損傷細(xì)胞的能力降低,但也可通過提高感染菌量而加大損傷。WT與ssal△分別感染口腔上皮細(xì)胞,ssal△感染組的GM-CSF、IL-1α、IL-1β、 RANTES、MIP-3α、IL-8的分泌量均較WT感染組降低(p0.05)。WT與ssal△分別感染陰道上皮細(xì)胞時(shí),ssa1△感染組在G-CSF、IL-1 α、MIP-3 α、IL-8細(xì)胞因子的產(chǎn)生量降低,與WT感染組相比有統(tǒng)計(jì)學(xué)差異(p0.05)。白念珠菌WT株感染兩種不同來源的上皮細(xì)胞時(shí),口腔上皮細(xì)胞組IL-1β、RANTES、 MIP-3 α、IL-8的分泌量比陰道上皮細(xì)胞組高(p0.05)。而在ssa1△感染兩種細(xì)胞時(shí),兩組細(xì)胞上述細(xì)胞因子產(chǎn)生量的對比無統(tǒng)計(jì)學(xué)差異(p0.05)�？谇簧掀ぜ�(xì)胞分別感染白念珠菌WT與ssa1△株2h后檢測通路蛋白,WT感染組p-ERK12及p-MKP1蛋白表達(dá)量較ssa1A感染組高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。在陰道上皮細(xì)胞分別感染W(wǎng)T與ssal△的蛋白檢測中,白念珠菌WT感染組p-ERK12蛋白及p-MKP1蛋白的表達(dá)量均較ssa1△感染后的表達(dá)量高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論：白念珠菌SSA1基因與白念珠菌菌絲延長的功能有關(guān)。SSA1基因的敲除,降低了白念珠菌對上皮細(xì)胞的損傷,也降低了白念珠菌對口腔上皮細(xì)胞及陰道上皮細(xì)胞炎性細(xì)胞因子的產(chǎn)生及信號(hào)通路的激活�？谇簧掀ぜ�(xì)胞及陰道上皮細(xì)胞在應(yīng)對白念珠菌Ssa1蛋白誘導(dǎo)的免疫反應(yīng)中,存在部位性差異。小結(jié)：白念珠菌感染口腔上皮細(xì)胞及陰道上皮細(xì)胞時(shí),兩種不同來源的上皮細(xì)胞激發(fā)的免疫效應(yīng)并不相同,其所分泌炎癥效應(yīng)因子及ERK炎癥通路蛋白表達(dá)方面存在差異,這可能是口腔、陰道兩部位抗白念珠菌結(jié)果差異的機(jī)制之一。通過敲除白念珠菌胞壁成分基因ALS3、SSA1的菌株,分別感染口腔上皮細(xì)胞及陰道上皮細(xì)胞,發(fā)現(xiàn)敲除后的菌株感染可以降低細(xì)胞產(chǎn)生的炎癥細(xì)胞因子及通路蛋白表達(dá),還可消除兩種細(xì)胞存在的部分差異,故白念珠菌ALS3、SSA1基因表達(dá)對誘導(dǎo)口腔及陰道上皮細(xì)胞抗白念珠菌感染免疫反應(yīng)的發(fā)生,有重要作用；對口腔、陰道兩部位抗白念珠菌感染免疫差異的發(fā)生也有重要作用,此實(shí)驗(yàn)結(jié)果可為口腔念珠菌感染及陰道念珠菌感染的防治工作研究提供更多的理論依據(jù)。
[Abstract]:Candida is a common opportunistic pathogenic fungus, in which Candida albicans is the main Candida albicans causing mucous infection in the oral and vaginal parts of the human body. It is known that mucous epithelial cells in the oral and vaginal sites are the first line of defense against Candida infection. Earlier studies found that epithelial cells resisted Candida albicans infection. It is necessary to activate inflammatory cell factors by activating inflammatory signaling pathway (such as ERK/JUK/p38 signaling pathway) and releasing inflammatory cytokines. In the light of clinical oral candidiasis, it is easier to cause systemic dissemination of infection in immunocompromised patients, but vaginal candidiasis is more likely to occur in the immune normal women. The epithelial cells in the same site may have different mechanisms for anti Candida albicans infection. The cell wall composition of Candida albicans, ALS3 and SSA1, is related to adhesion function. It plays an important role in the induction of immunization of Candida albicans in epithelial cells. Earlier studies have shown that knockout ALS3 can lower the secretion of oral epithelial cells to produce inflammation. The expression of cytokines and inflammatory signaling pathway proteins. However, there are few experimental studies on the effects of ALS3 knockout on the induced inflammatory cytokines induced by vaginal epithelial cells and the possible inflammatory pathways involved. There are few experimental studies on the effects of SSAl knockout on the inflammatory cytokines produced by epithelial cells and the possible inflammatory pathways involved. The experiment mainly studies the production of inflammatory cytokines induced by oral Candida albicans infection in oral epithelial cells and vaginal epithelial cells and the difference in molecular mechanism of ERK signaling protein activation. The possible mechanism of AIs3 protein in Candida albicans and the possible mechanism of Ssa1 protein in this immunization process. Immunological differences of Candida albicans against Candida albicans in vitro: To explore the similarities and differences between oral epithelial cells and vaginal epithelial cells in vitro infection of Candida albicans. Methods: oral epithelial cells (leuk-1), vaginal epithelial cells (VK2/E6E7) were cultured in vitro, and the infection of different strains of albicans (SC5314) was given. Differences in the expression of immune effector and pathway protein in non infected and Candida albicans infected epithelial cells. The cytotoxicity, inflammatory cytokine secretion and the difference of p-ERK1/2, p-MKP1 protein expression in Candida albicans infected with different epithelial cells. Results: oral and vaginal epithelial cells, in vitro culture form There are a lot of common features: the appearance of paving stone under inverted microscope, polygon, different size, irregular shape, round or oval nuclei. After dyed by the Rayleigh GIM composite dye, the cell can show a more typical cytoplasm pink, nuclear blue (black) color. Growth curve determination results, oral leuk- The growth and proliferation of 1 cells and vaginal VK2/E6E7 cells were different, and the proliferation rate of Leuk-1 cells was faster than that of VK2/E6E7 cells. The diameter of cells in the two groups was similar to the shape of "parabola" in the process of growth and proliferation, but the diameter of leuk-1 cells was still larger than that of VK2/ E6E7 cells. The cell damage test showed that the damage degree of the same cell was related to the amount of Candida albicans infection and the time of infection, that is, the damage degree was proportional to the amount of Candida albicans, and the two groups of cells were specific to the damage of different epithelial cells: in the condition of the same amount of infection, white Candida albicans The damage of Candida to the VK2/E6E7 of vaginal epithelial cells was heavy, but the damage to the leuk-1 of oral epithelial cells was lighter. The detection of inflammatory cytokines showed that Candida albicans in oral epithelial cells and vaginal epithelial cells stimulated GM-CSF, G-CSF, IL-1 a, IL-1 beta, RANTES, MIP-3 a, and IL-8 increased, but there was also a certain difference. The oral epithelial cells can produce a variety of cytokines in the uninfected state and produce more secretory quantities at low doses of Candida albicans, while most factors secrete most of the vaginal epithelial cells when they are not infected. Even after Candida albicans infection, the secretion of most cytokines is still small. Two kinds of cells appear. In the process of oral infection, the peak of IL-8, GM-CSF, MIP-3 alpha and RANTES appeared during the oral infection process, when the peak of Candida albicans infection / cell volume (MOI) =0.01, the peak of the secretion peak of G-CSF, IL-1 beta appeared at MOI=0.1, and the peak of IL-1 a appeared in MOI=1. And the vaginal epithelial cells were at the peak. During the infection process, when the peak of IL-8 beta GM-CSF secreted at MOI=0.01, the peak of G-CSF secreted at MOI=0.1, and the peak of IL-1 a, IL-1 beta and MIP-3 alpha appeared at MOI=1.IL-4 and IL-12 two cytokines, which did not show a significant dose dependence or regular trend of Candida albicans. In Candida albicans infection oral and vaginal After 2h, the expression of p-ERK1/2 and p-MKP1 protein were detected respectively. The expression of p-ERK1/2 and p-MKP1 protein in the white memory SC5314 infection group was higher than that of the control group. The difference was statistically significant (P0.05). Conclusion: Candida albicans infection in the oral and vaginal two different parts of the epithelial cells caused by cell damage and inflammation The immune response of Candida albicans against Candida albicans is different. Second the comparative study of the immunological mechanism of Candida albicans ALS3 knockout strains infected with oral and vaginal epithelial cells: To explore the production of Candida albicans ALS3 gene in Candida albicans to stimulate oral epithelial cells and vaginal epithelial cells The effect of anti Candida albicans immune response and the possible differences in immune mechanism. Methods: oral epithelial cells and vaginal epithelial cells were cultured in vitro, and the infection of Candida albicans SC5314 (WT strain) and Candida albicans ALS3 knockout (als3 delta) strains were given. The infection of Candida albicans and als3 delta strains were determined respectively. The cytotoxicity, inflammatory cytokines and the expression of ERK1/2, MKP1 pathway protein in different epithelial cells, compared with different strains, different strains, different sources of cell immune response. Results: the growth of the mycelium of Candida albicans ALS3 gene was not affected, and compared with the wild SC5314, there was no statistical difference (P0.05). But the comparison was compared with that of the wild SC5314 (P0.05). The results of WT and als3 delta infection of epithelial cells showed that the damage of als3 delta to cells was significantly reduced (P0.05), and the degree of cell damage was dose-dependent, that is, the greater the MOI value, the greater the damage to.Als3 delta infection of oral epithelial cells, GM-CSF, G-CSF, and the production of IL-1 alpha, IL-1 beta, MIP-3 alpha and IL-8 cell factors were lower than those of the infected group. After infection of vaginal epithelial cells, all the other cytokines except RANTES and MIP-3 a were lower than those in the WT group (P0.05). The results of WT strains respectively infected by two different sources of epithelial cells induced G-CSF, RANTES, MIP-3 alpha and IL-8, respectively, found that the production of these cytokines in the leuk-1 group of oral epithelial cells was higher than that of the vaginal cell VK2/E6E7 group. .05), while als3 delta infection, there was no statistical difference between the two groups of cell factors (P0.05). The expression of p-ERK1/2 protein in WT infection group and als3 delta infection group had no statistical difference (P0.05) after infection of Candida albicans WT and als3 delta 2H respectively, but the expression of p-MKP1 protein in WT infection group was higher than that of those in WT infection group. The expression of delta infection group was significant (P0.05). In the protein detection of VK2/E6E7 infection of Candida albicans WT and als3 Delta in vaginal epithelial cells, the expression of p-ERK1/2 protein and p-MKP1 protein in WT infection group were higher than that of als3 delta infection group, and the difference has the significance of unified planning (P0.05). Conclusion: ALS3 gene of Candida albicans The expression can induce a variety of cytokines in oral epithelial cells and vaginal epithelial cells. And the ability of als3 delta to stimulate two groups of cells to produce some inflammatory cytokines is different. In the process of inducing immune response in oral Candida albicans infected oral epithelial cells, A1s3 protein does not enhance the p-M by activating the ERK immune signal pathway. The expression of KP1 protein is involved in the immune response, and in the process of Candida albicans infected with vaginal epithelial cells, Als3 protein induces the enhancement of the expression of p-MKP1 protein by activating the ERK pathway to participate in the immune response. It shows that oral epithelial cells and vaginal epithelial cells exist in the immune response induced by Als3 protein of Candida albicans. A comparative study of the immunological mechanism of Candida albicans SSA1 knockout strains infected with oral and vaginal epithelial cells in Chapter third: To explore the role of the SSAl gene expression in oral Candida albicans and vaginal epithelial cells to stimulate the immune effect of Candida albicans in oral epithelial cells and vaginal epithelial cells. Using Candida albicans SC5314 (WT) and Candida albicans SSAl gene knockout strain (SSA1 delta), the changes in cell injury, inflammatory cytokine secretion, and pathway protein p-ERK1/2 and p-MKP1 expression in two different epithelial cells were measured respectively. The possible mechanisms were analyzed in contrast to the differences between different groups. The strain of Candida albicans SSAl knockout strain was cultured. After 2h, the length of mycelium was significantly shorter than that of WT strain (P0.05).Ssa1 Delta. After infection of epithelial cells 24h, the colony could not be formed. Only the scattered mycelium, after trypan blue was stained, the death cells were distributed along the mycelium. Compared with the result of the WT infected skin cells, the ability of SSA1 delta damaged cells was reduced. But by increasing the amount of infection bacteria, the damage of.WT and SSAL delta infection in oral epithelial cells, GM-CSF, IL-1 a, IL-1 beta, RANTES, MIP-3 a, IL-8 in SSAL delta infection group are lower than those of WT infection group (P0.05). The amount of production decreased, compared with the WT infection group (P0.05). When the WT strain of Candida albicans infected with two different sources of epithelial cells, the secretion of IL-1 beta, RANTES, MIP-3 a, IL-8 was higher than that of the vaginal epithelial cell group (P0.05) when the WT strain of Candida albicans was infected. The amount of cytokines produced above in the two groups was in the SSA1 delta infection of the two cells. There was no statistical difference (P0.05). Oral epithelial cells were infected with Candida albicans WT and SSA1 delta 2H respectively, and p-ERK12 and p-MKP1 protein expression in WT infection group were higher than that of ssa1A infection group, and the difference was statistically significant (P0.05). In the protein detection of WT and SSAL delta infection in vaginal epithelial cells, Candida albicans WT infection group The expression of K12 protein and p-MKP1 protein was higher than that of SSA1 delta infection. The difference was statistically significant (P0.05). Conclusion: the deletion of.SSA1 gene related to the function of SSA1 gene of Candida albicans and the extension of Candida albicans hyphae decreased the damage of Candida albicans to epithelial cells, and also reduced Candida albicans to oral epithelial cells and Yin. The production of inflammatory cytokines in the epithelial cells and the activation of the signal pathway. There is a regional difference between oral epithelial cells and vaginal epithelial cells in response to the immune response induced by Ssa1 protein of Candida albicans. Summary: when Candida albicans infected with oral epithelial cells and vaginal epithelial cells, two different sources of epithelial cells excite the epithelial cells. The effect of pestilence is not the same. There is a difference in the expression of inflammatory effect factor and ERK inflammatory pathway, which may be one of the mechanisms of oral and vaginal two parts of Candida albicans. By knocking out the strains of Candida albicans cell wall component gene ALS3 and SSA1, the infection of oral epithelial cells and vaginal epithelial cells, respectively, is found. The strain infection after knockout can reduce the expression of inflammatory cytokines and pathway proteins produced by the cells, and also eliminate the partial differences in the presence of two kinds of cells. Therefore, the expression of ALS3 and SSA1 gene of Candida albicans plays an important role in inducing the immune response of oral and vaginal epithelial cells against Candida albicans infection. Two parts of the oral cavity and vagina are resistant to the immune response to Candida albicans infection. Candida albicans infection also plays an important role in the occurrence of immune differences. The results can be used to prevent Candida infection and vaginal Candida infection.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R519.3

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