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  本文選題：膀胱腫瘤　切入點：BCG　出處：《山西醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:探討聯(lián)合應(yīng)用Ag85A與Ag85B DNA疫苗抗膀胱腫瘤的免疫效應(yīng)。 方法:采用體外培養(yǎng)的小鼠膀胱腫瘤細(xì)胞株BTT739,皮下注射T739小鼠的左后肢內(nèi)側(cè),成功構(gòu)建可移植性小鼠膀胱腫瘤模型。取48只,隨機數(shù)字表法隨機分為6組,即生理鹽水組、pcDNA3.1+組、卡介苗(BCG)組、pcDNA-Ag85A組、pcDNA-Ag85B組以及pcDNA—Ag85A+pcDNA—Ag85B組。荷瘤后第7天、14天和21天各肌注1次,末次肌注后7天處死小鼠,流式細(xì)胞儀檢測各組小鼠脾臟的CD4~+和CD8~+ T細(xì)胞亞群數(shù)量及所占百分比并計算CD4~+/CD8~+比值;酶聯(lián)免疫吸附測定(ELISA)法檢測小鼠血清中的干擾素γ(γ-IFN)分泌水平;定期測量腫瘤大小,計算腫瘤體積;剝離腫瘤稱重并作腫瘤組織病理學(xué)檢查。 結(jié)果:1.經(jīng)雙酶切鑒定,成功提取重組質(zhì)粒pcDNA-Ag85A及pcDNA-Ag85B。 2.CD4~+和CD8~+ T細(xì)胞亞群流式細(xì)胞學(xué)檢測結(jié)果:Ag85A與Ag85B DNA疫苗聯(lián)合免疫組CD4~+和CD8~+ T細(xì)胞亞群的數(shù)量及CD4~+/CD8~+的比值明顯高于DNA疫苗單獨免疫組,差異有顯著性(p＜0.05),但不及BCG組(p＜0.05)。 3.小鼠血清IFN-γ水平的ELISA檢測結(jié)果:Ag85A與Ag85B DNA疫苗聯(lián)合免疫組小鼠血清IFN-γ的水平明顯高于DNA疫苗單獨免疫組,差異有顯著性(p＜0.05),但不及BCG組(p＜0.05)。 4.腫瘤稱重:Ag85A與Ag85B DNA疫苗聯(lián)合免疫組小鼠的腫瘤重量明顯低于DNA疫苗單獨免疫組,差異有顯著性(p＜0.05),但高于BCG組(p＜0.05)。 5.腫瘤組織病理學(xué)變化:Ag85A與Ag85B DNA疫苗聯(lián)合免疫組腫瘤壞死(核濃縮、核碎裂、核溶解,胞漿紅染)較DNA疫苗單獨免疫組明顯,且瘤體周圍及瘤體內(nèi)炎細(xì)胞(中性粒細(xì)胞、淋巴細(xì)胞、單核細(xì)胞)浸潤:但BCG組腫瘤壞死更為明顯,瘤體周圍及瘤體內(nèi)有大量炎細(xì)胞浸潤。 結(jié)論:1.Ag85A與Ag85B DNA疫苗聯(lián)合免疫在抗膀胱腫瘤過程中具有明顯的免疫效應(yīng),且明顯優(yōu)于DNA疫苗單獨免疫,二者聯(lián)用具有協(xié)同增強免疫作用; 2.Ag85A與Ag85B DNA疫苗聯(lián)合免疫抗膀胱腫瘤效應(yīng)尚達(dá)不到BCG水平。
[Abstract]:Objective: to investigate the immunological effect of combined use of Ag85A and Ag85B DNA vaccine against bladder cancer. Methods: the mouse bladder tumor cell line BTT739 was cultured in vitro and injected subcutaneously into the medial left hind limb of T739 mouse. 48 mice were randomly divided into 6 groups. That is, normal saline group (pcDNA3.1), BCG (BCG) group, pcDNA-Ag85A group (pcDNA-Ag85B) and pcDNA-Ag85A pcDNA-Ag85B group. The number and percentage of CD4 ~ and CD8T lymphocyte subsets in spleen of each group were detected by flow cytometry and the ratio of CD4 ~ / / CD8 ~ ~ was calculated. The levels of interferon 緯 (緯 -IFNs) in serum of mice were detected by Elisa, and tumor size was measured regularly. Tumor volume was calculated, tumor weight was removed and histopathological examination was performed. Results 1. The recombinant plasmid pcDNA-Ag85A and pcDNA-Ag85B were successfully extracted by double enzyme digestion. 2. The number of CD4 ~ and CD8T subsets and the ratio of CD4 ~ / / CD8 ~ ~ ~ of CD4 ~ / CD8 ~ ~ were significantly higher in CD4 ~ and CD8- T cell subsets immunized with Ag85B DNA vaccine than those in DNA vaccine alone group, the difference was significant (P < 0.05), but less than that in BCG group (p < 0.05). 3. The serum IFN- 緯 level of mice immunized with Ag85B DNA vaccine was significantly higher than that of mice immunized with DNA vaccine alone (P < 0.05), but less than that of BCG group (P < 0.05). 3. The level of serum IFN- 緯 in mice immunized with Ag85B DNA vaccine was significantly higher than that in mice immunized with DNA vaccine alone (P < 0.05). 4. The tumor weight of mice immunized with Ag85B DNA vaccine was significantly lower than that of mice immunized with DNA vaccine alone (P < 0.05), but higher than that of BCG group (P < 0.05). 5. Histopathological changes of tumor tissues in the tumor necrosis group (nuclear concentration, nuclear fragmentation, nucleolysis, cytoplasmic red stain) in the combined immunization group with Ag85B DNA vaccine were significantly higher than those in the DNA vaccine alone group, and the inflammatory cells (neutrophils) around the tumor and in the tumor body. Lymphocyte, monocyte) infiltration: however, tumor necrosis was more obvious in BCG group, and a large number of inflammatory cells were infiltrated around the tumor and in the tumor body. Conclusion the combined immunization of Ag85B DNA vaccine with 1: 1 Ag85A vaccine has obvious immune effect in the process of anti-bladder tumor, and is obviously superior to that of DNA vaccine alone. The combination of the two vaccines can enhance the immunity synergistically. 2. The effect of Ag85A combined with Ag85B DNA vaccine on bladder tumor was not up to the level of BCG.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 楊登科,靳風(fēng)爍,江軍,張勇;分支桿菌Ag85B蛋白對小鼠膀胱癌抑癌效應(yīng)的研究[J];臨床泌尿外科雜志;2005年05期

2 武文森，尹克錚，，韓月明，張曉明，張東生，洪大落;小鼠可移植性膀胱移行細(xì)胞癌株（BTT739）的建立及實驗研究[J];中國腫瘤臨床;1996年10期

本文編號：1587889