【摘要】：疫苗佐劑的發(fā)展直接影響到新型疫苗的開發(fā)及應(yīng)用,為了開發(fā)出更安全、高效、經(jīng)濟(jì)以及更具有靶向性的佐劑,國(guó)內(nèi)外研究人員進(jìn)行了大量的研究。目前,鋁佐劑仍然是唯一在世界范圍內(nèi)被批準(zhǔn)用于人用疫苗的佐劑,它主要依賴儲(chǔ)存庫(kù)效應(yīng)和免疫刺激效應(yīng)的作用機(jī)制,能夠顯著提高疫苗在體內(nèi)的效果,經(jīng)過(guò)了幾十年的應(yīng)用,雖然出現(xiàn)部分輕微過(guò)敏反應(yīng),但總體來(lái)說(shuō)其安全性和有效性均得到了證實(shí)。 鋅是人體必需微量元素之一,人體生長(zhǎng)發(fā)育離不開鋅,體內(nèi)鋅水平的高低會(huì)對(duì)神經(jīng)、生殖和免疫系統(tǒng)產(chǎn)生影響。利用鋅與機(jī)體免疫系統(tǒng)之間的復(fù)雜關(guān)系,鋅療法被越來(lái)越廣泛地應(yīng)用于一些疾病的治療中。氫氧化鋅與氫氧化鋁理化性質(zhì)相近,在一定條件下,二者均能以穩(wěn)定的膠體溶液形式存在,由于鋁佐劑在臨床應(yīng)用中取得了良好的效果,因此將氫氧化鋅作為疫苗佐劑進(jìn)行的研究也非常具有潛力,在前期研究中,也初步證實(shí)了氫氧化鋅佐劑的安全性和有效性。在本研究中,參照鋁佐劑的制備方法,首先將可溶性鋅鹽葡萄糖酸鋅作為研究對(duì)象,利用其與低濃度氫氧化鈉溶液發(fā)生反應(yīng),制備出性質(zhì)穩(wěn)定質(zhì)地均勻的氫氧化鋅膠體溶液,之后再針對(duì)疫苗佐劑的要求對(duì)其理化性質(zhì)進(jìn)行改進(jìn),最終制得可以作為疫苗佐劑使用的氫氧化鋅膠體溶液。 三磷酸腺苷二鈉(ATP)是一種輔酶,有改善機(jī)體代謝的作用,同時(shí)又是體內(nèi)能量的主要來(lái)源。ATP能在某些免疫反應(yīng)中扮演重要角色,實(shí)驗(yàn)表明,ATP在與MgCl2復(fù)合使用后,可以對(duì)體內(nèi)IgG、IgA、IgM表達(dá)水平產(chǎn)生一定的影響,同時(shí),ATP是體內(nèi)一類重要的內(nèi)源性危險(xiǎn)信號(hào)因子,在缺氧、組織損傷、病變發(fā)生時(shí),多會(huì)伴隨周圍組織體液中ATP水平的變化,利用這一特點(diǎn),改變體內(nèi)ATP的水平在 一定條件下也能有效激活免疫系統(tǒng),并影響其效應(yīng)過(guò)程。因此,ATP也作為一類潛在疫苗佐劑得到進(jìn)一步的開發(fā)和研究。 復(fù)合佐劑是近年來(lái)佐劑研究的新熱點(diǎn),由于一些疫苗佐劑單獨(dú)作用效果并不理想,而將多種疫苗佐劑復(fù)合使用以后卻能達(dá)到讓人滿意的效果,因此關(guān)于復(fù)合佐劑的研究也不斷深入。本研究中,分別使用已經(jīng)得到廣泛運(yùn)用的鋁佐劑以及實(shí)驗(yàn)中制備的氫氧化鋅膠體佐劑,與ATP制備得到復(fù)合佐劑,再進(jìn)一步研究復(fù)合佐劑與鋁、鋅佐劑單獨(dú)作用的效果差異。實(shí)驗(yàn)中,以人用狂犬病疫苗作為抗原,分別設(shè)置不添加佐劑的抗原對(duì)照組、僅添加鋁、鋅佐劑的單獨(dú)佐劑組和分別添加了ATP-氫氧化鋁佐劑和ATP-氫氧化鋅佐劑的復(fù)合佐劑組,對(duì)ICR小鼠進(jìn)行免疫,在初次免疫后不同時(shí)間段通過(guò)間接ELISA法檢測(cè)小鼠血清中抗-狂犬病病毒IgG的滴度進(jìn)行免疫效果評(píng)價(jià)。 體液免疫效果評(píng)價(jià)結(jié)果表明,Zn(OH)2膠體與鋁佐劑類似,其作為佐劑單獨(dú)使用或與ATP聯(lián)合使用均能一定程度上增強(qiáng)機(jī)體針對(duì)狂犬病毒抗原的免疫應(yīng)答,雖然佐劑1次免疫組尚未達(dá)到抗原5次免疫組(抗原對(duì)照Ⅳ組)效果(P0.05),但抗體水平已經(jīng)基本高于抗原1-3次免疫組(抗原對(duì)照Ⅰ、Ⅱ、Ⅲ組)(P0.05),提示Zn(OH)2膠體佐劑單獨(dú)使用以及與ATP聯(lián)合使用均能增強(qiáng)狂犬病疫苗特異性體液免疫應(yīng)答,有一定的免疫增強(qiáng)作用。通過(guò)橫向?qū)Ρ瓤梢园l(fā)現(xiàn),氫氧化鋅膠體作為佐劑使用的效果已經(jīng)接近甚至超過(guò)鋁佐劑,加之鋅元素的毒副作用要遠(yuǎn)遠(yuǎn)低于鋁元素,鋅佐劑的開發(fā)應(yīng)用也有望克服鋁佐劑的應(yīng)用瓶頸,達(dá)到一個(gè)新的高度。
[Abstract]:The development of vaccine adjuvants has a direct impact on the development and application of new vaccines. In order to develop safer, more efficient, more economical and more targeted adjuvants, researchers at home and abroad have done a lot of research. The mechanism of response and immune stimulation can significantly improve the efficacy of the vaccine in vivo. After decades of application, although some slight allergic reactions have occurred, its safety and effectiveness have been confirmed in general.
Zinc is one of the essential trace elements in human body. Zinc can not be separated from the growth and development of human body. The level of zinc in the body will affect nervous, reproductive and immune system. Using the complex relationship between zinc and the immune system, zinc therapy is more and more widely used in the treatment of some diseases. Recently, both of them can exist in a stable colloidal solution under certain conditions. As aluminum adjuvant has achieved good results in clinical application, the study of zinc hydroxide as vaccine adjuvant has great potential. In the previous study, the safety and effectiveness of zinc hydroxide adjuvant were preliminarily confirmed. According to the preparation method of aluminium adjuvant, zinc gluconate, a soluble zinc salt, was used as the research object to prepare zinc hydroxide colloidal solution with stable and homogeneous properties by reacting with low concentration of sodium hydroxide solution. Then the physicochemical properties of zinc hydroxide colloidal solution were improved according to the requirements of vaccine adjuvant. Zinc hydroxide colloid solution for vaccine adjuvants.
ATP is a coenzyme that can improve metabolism and is also the main source of energy in the body. ATP can play an important role in some immune responses. The experiment shows that ATP combined with MgCl2 can affect the expression of IgG, IgA and IgM in vivo, and ATP is an important class in vivo. Endogenous risk factors, when hypoxia, tissue damage, lesions occur, are often associated with changes in the levels of ATP in the surrounding tissues and body fluids, using this feature to change the level of ATP in the body.
Under certain conditions, ATP can activate the immune system effectively and affect its effect process. Therefore, ATP has been further developed and studied as a potential vaccine adjuvant.
Complex adjuvant is a new hotspot in adjuvant research in recent years, because some vaccine adjuvants alone effect is not ideal, but after the use of multiple vaccine adjuvants can achieve satisfactory results, so the study of complex adjuvant is also in-depth. In this study, the use of aluminum adjuvant and solid adjuvant has been widely used. In the experiment, human rabies vaccine was used as antigen, and the antigen control group without adjuvant was set up, and the adjuvant group with aluminum and zinc adjuvant and the adjuvant group with ATP were added respectively. The ICR mice were immunized with the combined adjuvant of - aluminium hydroxide adjuvant and ATP-zinc hydroxide adjuvant. The titers of anti-rabies virus IgG in the serum of the mice were detected by indirect ELISA at different time after the first immunization.
The results of humoral immunity evaluation showed that Zn (OH) 2 colloid was similar to aluminum adjuvant, and its adjuvant alone or combined with ATP could enhance the immune response to rabies virus antigen to a certain extent. Although the adjuvant once immunized group had not reached the effect of antigen five times immunization group (antigen control group IV) (P 0.05), the antibody level had already been increased. The results showed that Zn (OH) 2 colloidal adjuvant alone and combined with ATP could enhance the specific humoral immune response of rabies vaccine, and had a certain immune enhancement effect. The development and application of zinc adjuvant is expected to overcome the bottleneck of the application of aluminum adjuvant and reach a new height.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 王建英;楊桂文;田雪文;李云龍;;免疫佐劑及其作用機(jī)制[J];山東師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2006年01期

2 李德娟;何巍;張睿;付琳娜;趙麗劍;王宇;金立杰;;氫氧化鋁佐劑配制工藝的優(yōu)化[J];中國(guó)生物制品學(xué)雜志;2010年10期

3 孫營(yíng);陳靜;陳少華;曹t，

本文編號(hào)：2208915