【摘要】：豬瘟是嚴(yán)重危害養(yǎng)豬業(yè)的一種病毒性傳染病,注射豬瘟疫苗是預(yù)防豬瘟的重要方法。目前常用的豬瘟疫苗佐劑副作用較強(qiáng),安全性和增強(qiáng)免疫效果有待提高,同時,還存在黏度高,注射使用不方便等問題。本文針對新發(fā)展的豬瘟活疫苗,研制安全性好、能夠有效增強(qiáng)免疫效果、使用方便的豬瘟活疫苗新型佐劑,并對O/W乳劑型豬瘟活疫苗佐劑的安全性和增強(qiáng)免疫效果進(jìn)行評價。對W/O/W乳劑型疫苗佐劑和聚合物凝膠佐劑的處方和制備工藝進(jìn)行了初步研究。開展的主要研究工作如下:(1)采用高壓均質(zhì)技術(shù)制備了O/W乳劑型豬瘟活疫苗佐劑,采用響應(yīng)面實驗設(shè)計優(yōu)化了豬瘟活疫苗佐劑的處方和制備工藝,并對其穩(wěn)定性、粒徑、多分散指數(shù)(PDI)及Zeta電位進(jìn)行表征。優(yōu)化處方和工藝制備的O/W型疫苗佐劑平均粒徑為100.4 nm,PDI為0.147,Zeta電位為-28.7 mV,離心穩(wěn)定性實驗表明其穩(wěn)定性良好。(2)進(jìn)行了O/W乳劑型豬瘟活疫苗佐劑小鼠安全性實驗。結(jié)果表明,注射佐劑組小鼠體重和生存狀態(tài)與空白對照組相比較沒有差異,沒有出現(xiàn)由佐劑引起的死亡或者明顯的局部和全身不良反應(yīng)。兩周后解剖小鼠,檢查發(fā)現(xiàn)其注射部位及心、肝、脾、肺、腎均正常,沒有中毒癥狀。表明研制的O/W型豬瘟活疫苗佐劑安全性良好。(3)將O/W乳劑型疫苗佐劑與豬瘟疫苗混合后,肌肉注射免疫小鼠,進(jìn)行佐劑增強(qiáng)免疫效果評價。實驗結(jié)果表明,佐劑組小鼠血清中抗豬瘟(CSFV)抗體水平與對照組有顯著差異,抗體水平能夠持續(xù)較長的時間,表明O/W乳劑型豬瘟活疫苗佐劑能夠有效誘導(dǎo)機(jī)體產(chǎn)生體液免疫應(yīng)答;佐劑組小鼠血清中IL-4、IL-6和IFN-γ的水平均有明顯提高,表明該疫苗佐劑能夠誘導(dǎo)機(jī)體產(chǎn)生細(xì)胞免疫應(yīng)答。(4)采用高速分散結(jié)合高壓勻質(zhì)技術(shù)制備W/O/W乳劑型疫苗佐劑,對其處方和制備工藝進(jìn)行了研究,通過篩選確定了乳化性能好的復(fù)合乳化劑和高壓勻質(zhì)制備工藝條件,優(yōu)化制備的W/O/W乳劑型疫苗佐劑穩(wěn)定性好,黏度適中。(5)開展了聚合物凝膠佐劑的處方和制備工藝研究,對聚合物凝膠基質(zhì)、乳化劑、油相以及高壓勻質(zhì)工藝進(jìn)行了篩選和優(yōu)化,優(yōu)化處方和工藝制備的聚合物凝膠佐劑平均粒徑為221.6 nm,PDI為0.114,Zeta電位為-29.7 mV,穩(wěn)定性良好。
[Abstract]:Swine fever (CSF) is a viral infectious disease which seriously endangers the swine industry. Injection of CSFV vaccine is an important method to prevent swine fever. At present, the adjuvant of classical swine fever vaccine has strong side effects, safety and immune enhancement effect need to be improved, at the same time, there are some problems such as high viscosity, inconvenient injection and so on. In this paper, a new adjuvant for swine fever vaccine is developed, which is safe, effective and easy to use. The safety and immune enhancement of O / W emulsion adjuvant of swine fever vaccine were evaluated. The formulation and preparation of W/O/W emulsion vaccine adjuvant and polymer gel adjuvant were studied. The main research works are as follows: (1) the O / W emulsion adjuvant of hog cholera vaccine was prepared by high pressure homogenization technique. The formulation and preparation process of live vaccine adjuvant of hog fever were optimized by response surface experiment, and the stability of the adjuvant was analyzed. Particle size, polydispersity index (PDI) and Zeta potential were characterized. The average particle size of the adjuvant prepared by optimized formulation and process is 100.4 nm,PDI, and the Zeta potential of the adjuvant is -28.7 mV,. Centrifugation stability test showed that the stability was good. (2) the safety test of adjuvant of O / W emulsion CSFV vaccine was carried out in mice. The results showed that there was no difference in body weight and survival status between the adjuvant group and the blank control group. There was no death caused by adjuvant or obvious local and systemic adverse reactions. Two weeks later, the mice were dissected and the injection site, heart, liver, spleen, lung and kidney were normal. The results showed that the adjuvant of O / W type swine fever vaccine was safe. (3) the adjuvant was injected intramuscularly to immunize mice with the adjuvant of O / W emulsion vaccine and swine fever vaccine, and the adjuvant was used to evaluate the effect of adjuvant. The results showed that the level of anti-CSFV (CSFV) antibody in the adjuvant group was significantly different from that in the control group, and the antibody level could last for a long time. The results showed that the adjuvant of O / W emulsion CSFV vaccine could induce humoral immune response effectively. The serum levels of IL-4,IL-6 and IFN- 緯 in the adjuvant group were significantly increased. The results showed that the adjuvant could induce cellular immune response. (4) W/O/W emulsion vaccine adjuvant was prepared by high speed dispersion and high pressure homogenization. The formulation and preparation process of the adjuvant were studied. The preparation conditions of composite emulsifier with good emulsifying performance and high pressure homogenization were determined by screening, and the adjuvant of W/O/W emulsion vaccine prepared by optimization was stable. (5) the formulation and preparation of polymer gel adjuvant were studied. The polymer gel matrix, emulsifier, oil phase and high pressure homogenization process were screened and optimized. The average particle size of the polymer gel adjuvant prepared by the optimized formulation and process was 221.6 nm,PDI and the Zeta potential was -29.7 mV,. The average particle size of the polymer gel adjuvant was -29.7 mV,.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張智明;張虹;畢艷蘭;孫曉洋;徐學(xué)兵;;旋轉(zhuǎn)流變儀在油脂研究中的應(yīng)用[J];中國油脂;2013年09期

2 劉晶;吳衍嶺;;豬瘟疫苗免疫過程中需要注意的問題[J];山東畜牧獸醫(yī);2012年11期

3 劉文麗;焦緯洲;劉有智;李靜;楊森;;W/O/W型多重乳液的穩(wěn)定性和應(yīng)用研究進(jìn)展[J];日用化學(xué)工業(yè);2012年05期

4 白成友;陳天澤;鄭玉晏;謝曉燕;謝君慶;;豬瘟疫苗的研究現(xiàn)狀及正確使用方法[J];吉林農(nóng)業(yè);2012年07期

5 高澤乾;孫建男;武煒;李成會;;獸用免疫佐劑的研究進(jìn)展[J];唐山師范學(xué)院學(xué)報;2012年02期

6 張曉艷;趙凱;侯志勇;陳剛;蒿連微;;殼聚糖及其衍生物作為免疫佐劑的研究進(jìn)展[J];中國新藥雜志;2012年02期

7 任銳;呂雪峰;孟憲奎;;獸用疫苗佐劑的研究及應(yīng)用現(xiàn)狀[J];吉林畜牧獸醫(yī);2011年05期

8 厙大亮;李冬;;新型獸用疫苗佐劑的研究進(jìn)展[J];中國獸藥雜志;2011年04期

9 何海勇;賁虎;梁興杰;;納米技術(shù)在疫苗佐劑研制中的應(yīng)用[J];東南大學(xué)學(xué)報(醫(yī)學(xué)版);2011年01期

10 黃興;岳華;;納米佐劑的研究進(jìn)展[J];中國畜牧獸醫(yī);2010年09期

相關(guān)碩士學(xué)位論文 前2條

1 錢歡;一種新型免疫佐劑對豬瘟疫苗免疫增強(qiáng)效果的研究[D];上海交通大學(xué);2012年

2 劉甜甜;新型免疫佐劑的研制及亞單位佐劑疫苗的免疫效力檢驗[D];中國海洋大學(xué);2012年

，

本文編號：2312401