發(fā)布時間：2018-11-08 10:13

【摘要】： 目的:研究BCG-CpG-DNA的免疫毒性作用,為BCG-CpG-DNA臨床前安全性評價(jià)提供依據(jù)。 方法:采用異常毒性試驗(yàn)、急性毒性試驗(yàn)、28天毒性試驗(yàn)初步觀察BCG-CpG-DNA毒性;并通過計(jì)算免疫器官臟器系數(shù),檢測淋巴細(xì)胞轉(zhuǎn)化能力、細(xì)胞因子釋放(Elospot法)、T細(xì)胞亞群變化(免疫熒光法)、NK細(xì)胞殺傷活性(LDH法),外周血抗雙鏈DNA自身抗體水平(Elisa法),與溶劑對照組比較,進(jìn)一步觀察BCG-CpG-DNA重復(fù)給藥對小鼠免疫功能的影響。 結(jié)果:(1)異常毒性試驗(yàn):實(shí)驗(yàn)期間,所有動物一般行為活動、外觀體征、進(jìn)食和飲水活動等均正常;所有動物全部健康存活;動物均有體重增加;大體解剖肉眼觀察各臟器均無異常。(2)急性毒性試驗(yàn):實(shí)驗(yàn)期間,所有動物一般行為活動、外觀體征、進(jìn)食和飲水活動等均正常;所有動物全部健康存活;動物均有體重增加;大體解剖,臟器無病變,各組脾重?zé)o統(tǒng)計(jì)學(xué)差異(P0.05)。(3)28天毒性試驗(yàn):①外觀體征、體重觀察:在給藥期間及恢復(fù)期,實(shí)驗(yàn)組和對照組動物均活動正常,進(jìn)食敏捷、毛貼身有光澤,糞便正常;動物體重各組間無統(tǒng)計(jì)學(xué)差異(P0.05)。②血液學(xué)指標(biāo):4次檢測結(jié)果表明,BCG-CpG-DNA主要影響小鼠白細(xì)胞總數(shù)、淋巴細(xì)胞絕對值、中性粒細(xì)胞絕對值及相應(yīng)的百分比,對其他指標(biāo)無明顯影響。③血生化學(xué)指標(biāo):4次檢測結(jié)果表明,BCG-CpG-DNA對小鼠外周血生化指標(biāo)無明顯影響。④大體解剖與臟器系數(shù):大體解剖觀察,各組動物的主要器官均未見明顯病理性改變;免疫中期即免疫后10天,BCG-CpG-DNA中、高劑量組脾臟重量系數(shù)高于溶劑對照組,有顯著性差異(P=0.03,0.05);免疫末期即末次免疫后3天,BCG-CpG-DNA低、中、高劑量組脾臟重量系數(shù)高于溶劑對照組,有顯著性差異(P=0.009,0.007,0.007);恢復(fù)期即免后21天,BCG-CpG-DNA低、中、高劑量組脾臟重量系數(shù)、肝重系數(shù)、胸腺重系數(shù)和腎臟重系數(shù),各劑量組和溶劑對照組均無統(tǒng)計(jì)學(xué)差異(P0.05);⑤局部刺激反應(yīng):在給藥期間及恢復(fù)期,實(shí)驗(yàn)組和對照組動物的給藥部位表觀無異常反應(yīng),大體解剖后,給藥部位也無異常反應(yīng)。(4)免疫功能檢測:BCG-CpG-DNA重復(fù)給藥,劑量分別達(dá)到0.7mg、1.75mg和3.5mg,對細(xì)胞免疫功能的影響主要有:增強(qiáng)淋巴細(xì)胞轉(zhuǎn)化功能;降低中、高劑量組CD3+T細(xì)胞含量;提高體內(nèi)分泌IFN-γ和IL-4的細(xì)胞數(shù)二者的比例;增強(qiáng)NK細(xì)胞殺傷能力,不同實(shí)驗(yàn)組與對照組差異均有顯著意義(P0.05);經(jīng)過三周恢復(fù)期,變化的指標(biāo)均恢復(fù)正常。 結(jié)論:BCG-CpG-DNA在異常毒性試驗(yàn)、急性毒性試驗(yàn)、28天毒性試驗(yàn)均未見明顯毒性反應(yīng);BCG-CpG-DNA重復(fù)給藥累計(jì)達(dá)0.7mg、1.75mg和3.5mg的情況下,小鼠表現(xiàn)為免疫功能增強(qiáng),但對免疫器官和免疫功能無不良影響,無免疫毒性作用,安全性良好。
[Abstract]:Objective: to study the immunotoxicity of BCG-CpG-DNA and to provide evidence for pre-clinical safety evaluation of BCG-CpG-DNA. Methods: abnormal toxicity test, acute toxicity test and 28 days toxicity test were used to observe the toxicity of BCG-CpG-DNA. The lymphocyte transformation ability and cytokine release were measured by calculating the organ coefficient of immune organs. The changes of), T cell subsets were detected by Elospot method (LDH assay), and the cytotoxicity of), NK cells by immunofluorescence assay (LDH method). The level of anti-double-stranded DNA autoantibodies in peripheral blood (Elisa method) was compared with that of solvent control group to observe the effect of repeated administration of BCG-CpG-DNA on the immune function of mice. Results: (1) abnormal toxicity test: during the experiment, all animals were normal in general behavior, appearance, eating and drinking, all animals were healthy and survived, all animals had weight gain. (2) Acute toxicity test: during the course of the experiment, all animals were normal in general behavior, appearance, eating and drinking water, all animals survived in health. All animals gained weight; Gross anatomy, viscera no lesion, spleen weight of each group (P0.05). (3) 28 days toxicity test: 1 physical signs, body weight observation: during the administration and recovery period, the experimental group and the control group were all normal activity. Eat quickly, hair close body luster, feces normal; There was no significant difference in body weight among the groups (P0.05). 2 Hematological indexes: the results showed that BCG-CpG-DNA mainly affected the total number of white blood cells, the absolute value of lymphocytes, the absolute value of neutrophils and the corresponding percentage. The results showed that BCG-CpG-DNA had no significant effect on the biochemical indexes of peripheral blood of mice. 4 gross anatomy and organ coefficient: gross anatomical observation. No obvious pathological changes were found in the main organs of the animals in each group. In BCG-CpG-DNA, the spleen weight coefficient of high dose group was higher than that of solvent control group (P < 0. 03). The spleen weight coefficient of the middle and high dose group was significantly higher than that of the solvent control group (P0. 009 0. 007 ~ 0. 007). In the recovery period, 21 days after immunity, there was no significant difference in spleen weight coefficient, liver weight coefficient, thymus weight coefficient and kidney weight coefficient between the low, middle and high dose groups of BCG-CpG-DNA (P0.05). (5) Local stimulation: during administration and convalescence, there was no apparent abnormal reaction in the administration site of the experimental group and the control group, and there was no abnormal response in the administration site after gross anatomy. (4) Immunologic function test: repeated administration of BCG-CpG-DNA; The doses of 0.7 mg and 3.5 mg were 1.75 mg and 3.5 mg, respectively. The main effects on the cellular immune function were as follows: enhancing the lymphocyte transformation function; Reducing the content of CD3 T cells in high dose group, increasing the proportion of IFN- 緯 and IL-4 secreting cells in the body, enhancing the killing ability of NK cells, there were significant differences between the different experimental groups and the control group (P0.05). After three weeks of convalescence, the index of change returned to normal. Conclusion: there were no obvious toxic reactions in abnormal toxicity test, acute toxicity test and 28 day toxicity test of BCG-CpG-DNA. Under the condition of repeated administration of BCG-CpG-DNA (0.7mg / kg) and 3.5mg (1.75mg), the mice showed enhanced immune function, but had no adverse effect on immune organs and immune function, and had no toxic effect and good safety.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 梁春來;賈旭東;;免疫毒性評價(jià)方法研究進(jìn)展[J];毒理學(xué)雜志;2011年04期

2 霍艷;艾文超;聞鎳;沈連忠;王軍志;李波;;TDAR在藥物臨床前毒理學(xué)研究中的應(yīng)用[J];中國藥事;2011年07期

3 王昭;王威;;二VA英的免疫毒性作用研究進(jìn)展[J];細(xì)胞與分子免疫學(xué)雜志;2011年07期

4 李文;林育純;陳慧峰;羅潔;李曉杰;林麗娜;胡耀明;張樹江;陳雯;林忠寧;;鋅影響鎘對人未成熟樹突狀細(xì)胞毒性作用[J];中國職業(yè)醫(yī)學(xué);2011年03期

5 余明東;袁萍;何杰穎;文紅玲;王友良;李東陽;;全氟辛烷磺酸對SD大鼠的免疫損傷作用[J];中南醫(yī)學(xué)科學(xué)雜志;2011年02期

6 張愛華;董明;潘巧裕;王俊;吳邦華;黃淑蓮;;八極桿碰撞反應(yīng)池-電感耦合等離子體質(zhì)譜法測定尿中鎳和鉻[J];理化檢驗(yàn)(化學(xué)分冊);2011年05期

7 高峰;張琨;吳曉剛;宋昕恬;張晶瑩;;鄰苯二甲酸二(2-乙基己基)酯毒性作用[J];職業(yè)與健康;2011年17期

8 王洋;;二惡英危害談[J];中國標(biāo)準(zhǔn)導(dǎo)報(bào);1999年04期

9 喬果果;張志紅;董潔;邱勇;郭麗麗;白楊;宋劍;陳剛;;交通相關(guān)的PM_(2.5)亞急性染毒對大鼠肺泡灌洗液和血清細(xì)胞因子水平的影響[J];毒理學(xué)雜志;2011年04期

10 安虹;鄒廣迅;;黃曲霉素毒性效應(yīng)機(jī)制的研究進(jìn)展[J];安徽農(nóng)業(yè)科學(xué);2011年24期

相關(guān)會議論文 前10條

1 呂琪妍;萬斌;郭良宏;;多溴聯(lián)苯醚的免疫毒性效應(yīng)和機(jī)制研究[A];第六屆全國環(huán)境化學(xué)大會暨環(huán)境科學(xué)儀器與分析儀器展覽會摘要集[C];2011年

2 甘燕;李俊華;劉洪波;劉茂玲;張仁利;高世同;黃達(dá)娜;耿藝介;吳少庭;;抗SARS病毒重組BCG-M疫苗的構(gòu)建及其免疫學(xué)研究[A];全國人畜共患病學(xué)術(shù)研討會論文集[C];2006年

3 甘燕;吳少庭;李俊華;劉洪波;黃達(dá)娜;余新炳;;抗SARS病毒重組BCG-M疫苗的構(gòu)建及其免疫學(xué)研究[A];全國寄生蟲學(xué)與熱帶醫(yī)學(xué)學(xué)術(shù)研討會論文集[C];2006年

4 胡琳潔;郭曉雅;侯美娜;邵成;史皆然;;評價(jià)空腸彎曲菌外膜蛋白PEB1介導(dǎo)的重組BCG與上皮細(xì)胞的結(jié)合能力[A];中華醫(yī)學(xué)會呼吸病學(xué)年會——2011（第十二次全國呼吸病學(xué)學(xué)術(shù)會議）論文匯編[C];2011年

5 Michael Graner;;Evaluation of performance of an In-house Interfer on-gamma Elispot assay for Latent Tuberculosis in BCG Vaccinated individuals[A];全國第3屆中西醫(yī)結(jié)合傳染病學(xué)術(shù)會議暨中國中西醫(yī)結(jié)合學(xué)會傳染病專業(yè)委員會第2屆委員會議論文匯編[C];2010年

6 李俊華;吳少庭;翁亞彪;甘燕;劉洪波;劉茂鈴;張仁利;高世同;黃達(dá)娜;耿藝介;;弓形蟲GRA4基因和SAG2基因的重組BCG疫苗對小鼠免疫效果的比較研究[A];全國寄生蟲學(xué)與熱帶醫(yī)學(xué)學(xué)術(shù)研討會論文集[C];2006年

7 童建;;農(nóng)藥免疫毒性研究進(jìn)展[A];中國毒理學(xué)會第三屆全國學(xué)術(shù)會議論文（摘要）集[C];2001年

8 鐘賽賢;賈山;余貴英;;鎘的免疫毒性與IL-2、CAMP影響的實(shí)驗(yàn)研究[A];中國毒理學(xué)會第三屆全國學(xué)術(shù)會議論文（摘要）集[C];2001年

9 呂琪妍;萬斌;郭良宏;;全氟辛烷磺酸基化合物的免疫毒性效應(yīng)和機(jī)制研究[A];中國化學(xué)會第28屆學(xué)術(shù)年會第2分會場摘要集[C];2012年

10 馬t，

本文編號：2318163