本文選題：天然抗體　切入點：白念珠菌　出處：《第四軍醫(yī)大學(xué)》2008年碩士論文

【摘要】： 天然抗體(natural antibodies, NAbs)是指正常機體在沒有任何抗原刺激的情況下由胚系基因編碼的,正常機體產(chǎn)生的針對一種或多種自身和(或)外源性抗原的抗體,具有多反應(yīng)性和低親和力的特征。自NAbs被發(fā)現(xiàn)以來,人們對其可能具有的功能進行了一系列的深入研究。研究結(jié)果表明,NAbs是構(gòu)成機體天然免疫系統(tǒng)的重要組成部分,在腫瘤監(jiān)視、自身免疫、異體排斥、抗感染等方面均發(fā)揮著重要的作用。 近年來,隨著抗生素、皮質(zhì)類固醇激素、免疫抑制劑等的廣泛應(yīng)用,以及HIV感染者增加、異種移植術(shù)的開展、腫瘤發(fā)病率的上升,真菌感染呈明顯上升趨勢,特別是白念珠菌引起的感染明顯上升,越來越引起人們的重視。人們逐漸認識到體液免疫在抗真菌感染中起著重要作用,而天然體液免疫又構(gòu)成了機體抵御外來感染的第一道防線,天然抗體的作用逐漸引起人們的重視。雖然有部分事實表明天然抗體可能具有抗真菌免疫的作用,但還缺乏直接的證據(jù),為此倍受醫(yī)學(xué)界學(xué)者的廣泛關(guān)注。 本課題組在國家自然科學(xué)基金(30330510、30300318, 30330550, 30425015)的支持下,應(yīng)用未免疫小鼠B細胞與骨髓瘤細胞融合,成功篩選到了3株分泌IgM型天然抗體的雜交瘤細胞,它們的可變區(qū)基因與胚系基因高度同源,其中以天然抗體IgM 3B4與抗原的結(jié)合活性最好,并且已經(jīng)證實在體外天然抗體IgM 3B4可以調(diào)理吞噬細胞對金黃色葡萄球菌的吞噬作用和天然抗體IgM 3B4可以與白念珠菌孢壁的糖蛋白抗原結(jié)合。本研究在前期工作的基礎(chǔ)上利用純化得到的天然抗體IgM 3B4,在體外以白念珠菌為對象,初步探討天然抗體IgM 3B4抗真菌感染的作用機制。 本研究利用純化得到的天然抗體IgM 3B4在體外證實:1在體外,不同濃度的天然抗體IgM 3B4可以抑制白念珠菌芽管生成,2在體外,天然抗體IgM 3B4可以抑制白念珠菌與宿主細胞之間的粘附;3采用ELISA和RT-PCR方法檢測了天然抗體IgM 3B4在體外對感染了白念珠菌的臍靜脈血管內(nèi)皮細胞IL-6、IL-8、ICAM-1和VCAM-1表達和分泌的影響,證實天然抗體IgM 3B4明顯促進了臍靜脈血管內(nèi)皮細胞上述細胞因子的表達和分泌。從而證實了天然抗體IgM 3B4抵抗白念珠菌感染的能力與其抑制白念珠菌芽管生成、抑制白念珠菌與宿主細胞之間的粘附以及促進臍靜脈血管內(nèi)皮細胞IL-6、IL-8、ICAM-1和VCAM-1的表達和分泌有關(guān)。
[Abstract]:Natural antigens (NAbs) are antibodies that are encoded by embryogenic genes in normal organisms without any antigenic stimulation, and are produced against one or more of their own and / or exogenous antigens. Since the discovery of NAbs, a series of in-depth studies have been carried out on its possible functions. The results show that NAbs is an important part of the innate immune system. It plays an important role in tumor surveillance, autoimmunity, allograft rejection and anti-infection. In recent years, with the widespread use of antibiotics, corticosteroids, immunosuppressants, and the increase of HIV infection, xenotransplantation, the incidence of tumor and fungal infection have increased significantly. In particular, the infection caused by Candida albicans has increased significantly and attracted more and more attention. People have come to realize that humoral immunity plays an important role in antifungal infections. And natural humoral immunity constitutes the first line of defense against foreign infections, and the role of natural antibodies has gradually attracted people's attention. Although some facts have shown that natural antibodies may have the effect of antifungal immunity, However, the lack of direct evidence has attracted wide attention from medical scholars. With the support of the National Natural Science Foundation of China, 30330510C30300318,30330550,30425015, three hybridoma cells secreting IgM type natural antibodies were successfully screened by fusion of unimmunized B cells with myeloma cells. Their variable region genes are highly homologous to embryogenic genes, and the natural antibody IgM 3B4 has the best binding activity to antigens. It has been proved that natural antibody IgM 3B4 can regulate phagocytosis of phagocytes to Staphylococcus aureus in vitro and that natural antibody IgM 3B4 can bind to glycoprotein antigen of Candida albicans spore wall. Based on the purified natural antibody IgM 3B4, Candida albicans was used in vitro. To explore the mechanism of natural antibody IgM 3B4 in antifungal infection. In this study, the purified natural antibody IgM 3B4 was used to confirm that IgM 3B4 of different concentrations could inhibit the formation of Candida albicans bud tube in vitro. Natural antibody IgM 3B4 could inhibit adhesion between Candida albicans and host cells. The effect of natural antibody IgM 3B4 on the expression and secretion of IL-8 ICAM-1 and VCAM-1 in umbilical vein endothelial cells infected with Candida albicans was detected by ELISA and RT-PCR in vitro. It was proved that the natural antibody IgM 3B4 significantly promoted the expression and secretion of the above cytokines in the endothelial cells of the umbilical vein, thus confirming the ability of the natural antibody IgM 3B4 to resist the infection of Candida albicans and to inhibit the formation of the germ tubes of Candida albicans. The inhibition of the adhesion between Candida albicans and host cells and the promotion of the expression and secretion of ICAM-1 and VCAM-1 in endothelial cells of umbilical vein (HUVEC) were related to the inhibition of the adhesion between Candida albicans and host cells.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R392

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