布鲁氏菌病温敏凝胶活疫苗(S2株)对小鼠不同黏膜免疫途径的免疫效果研究
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摘要
为了比较布鲁氏菌病温敏凝胶疫苗经不同黏膜途径免疫的效果,本研究采用BABL/c小鼠为实验动物模型,制备了用于黏膜免疫的布鲁氏菌病温敏凝胶活疫苗(S2株),比较了该疫苗与常规疫苗黏膜免疫(阴道灌注、点眼、口服、直肠灌注)后的体表排菌,研究了黏膜免疫与注射免疫的抗体反应规律差异及保护效果。结果显示:凝胶疫苗能够在34℃～36℃完成由液态到凝胶态的相转变,适合所选小鼠黏膜途径的免疫;凝胶疫苗从免疫后第2 d开始停止排菌,常规疫苗的排菌持续到免疫后第4 d;黏膜免疫小鼠抗体效价比注射免疫低25%～80%,并且在免疫后45 d左右微量试管凝集试验(MSAT)结果显示前者抗体效价即下降至1∶50以下或接近1∶50,下降速度显著快于注射免疫;对阴道灌注、点眼、口服、直肠灌注及注射免疫的小鼠攻毒结果显示对照组小鼠的出菌指数为6.47,免疫组的单位保护(对照出菌指数-免疫出菌指数)分别为3.89、3.81、3.32、3.25、4.08,其中阴道灌注、点眼2种黏膜免疫小鼠的单位保护更接近注射免疫的单位保护(4.08)。研究结果提示,凝胶疫苗比常规疫苗在免疫后更可以显著减少体表排菌,抗体持续时间更短、使得对疫病的诊断干扰更低,此外,凝胶疫苗免疫也具有接近注射免疫的单位保护。因此,采用布鲁氏菌病凝胶疫苗进行黏膜免疫更有利于布鲁氏菌病的鉴别诊断。
To compare the effects of brucellosis thermos-sensitive gel vaccine on different mucosal surface vaccinations in BABL/c mice, a live brucellosis thermosensitive gel vaccine strain S2(S2 gel vaccine) was prepared and compared with conventional vaccine for mucosal immunity via different mucosal surface including, eye-drop, oral administration, vaginal and rectal perfusion, which were evaluated by the surface reaction to the vaccine bacteria, the difference in antibody response between mucosal immunity and injection immunization as well as the protection test. The results showed that the S2 gel vaccine was able to complete the phase transition from liquid to gel state at 34℃ to 36 ℃, which was suitable for the mucosal immunization to the mice. The vaccine bacterial shedding stoped at 2 days post immuniztion(PI) with the S2 gel vaccine, but it stoped at 4 days PI with the conventional vaccine. The antibody levels in the mice of mucosal immunization groups were 25% to 80% lower than that of the mice in injection immunized groups, and the antibody titers detected with MSAT droped below 1:50 or close to 1 ∶50 after45 days of immunization, and the rate of decline was significantly faster. Moreover, the mice were challenged post immunizing by eye-drop, oral administration, vaginal and rectal perfusion, as well as injection immunization, respectively. The results showed that the bacterial index of the control group was 6.47, and the unit protections of the immunized groups(control bacterial index-immunization index) were 3.89, 3.81, 3.32, 3.25 and 4.08, respectively. The unit protection of immunization was closer to the unit protection of injection immunization(4.08). The results suggested that the S2 gel vaccine induced a shorter duration of antibody than the traditional vaccine, a lower interference to the diagnosis, and a near-injection immunity. Therefore, mucosal immunization with a brucellosis gel vaccine is more conducive to the differential diagnosis of brucellosis.
To compare the effects of brucellosis thermos-sensitive gel vaccine on different mucosal surface vaccinations in BABL/c mice, a live brucellosis thermosensitive gel vaccine strain S2(S2 gel vaccine) was prepared and compared with conventional vaccine for mucosal immunity via different mucosal surface including, eye-drop, oral administration, vaginal and rectal perfusion, which were evaluated by the surface reaction to the vaccine bacteria, the difference in antibody response between mucosal immunity and injection immunization as well as the protection test. The results showed that the S2 gel vaccine was able to complete the phase transition from liquid to gel state at 34℃ to 36 ℃, which was suitable for the mucosal immunization to the mice. The vaccine bacterial shedding stoped at 2 days post immuniztion(PI) with the S2 gel vaccine, but it stoped at 4 days PI with the conventional vaccine. The antibody levels in the mice of mucosal immunization groups were 25% to 80% lower than that of the mice in injection immunized groups, and the antibody titers detected with MSAT droped below 1:50 or close to 1 ∶50 after45 days of immunization, and the rate of decline was significantly faster. Moreover, the mice were challenged post immunizing by eye-drop, oral administration, vaginal and rectal perfusion, as well as injection immunization, respectively. The results showed that the bacterial index of the control group was 6.47, and the unit protections of the immunized groups(control bacterial index-immunization index) were 3.89, 3.81, 3.32, 3.25 and 4.08, respectively. The unit protection of immunization was closer to the unit protection of injection immunization(4.08). The results suggested that the S2 gel vaccine induced a shorter duration of antibody than the traditional vaccine, a lower interference to the diagnosis, and a near-injection immunity. Therefore, mucosal immunization with a brucellosis gel vaccine is more conducive to the differential diagnosis of brucellosis.
引文
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[9]管晓燕,李敏,刘建国,等.温敏型生物降解水凝胶载体防龋基因疫苗p VAX1-spap/A经不同途径免疫新西兰大白兔的实验研究[J].口腔医学研究,2014,10:28.
[10]翟良,王霄旸,王昊欣,等.蛋白载体温敏凝胶制备及释放度研究[J].中国兽药杂志,2014,48(5):32-35.
[11]Juan E U,Susana L,Rodolfo A U.Evaluation of Brucella abortus phosphoglucomutase(pgm)mutant as a new live rough-phenotype vaccine[J].Infect Immun,2003,71:6264.
[12]李晓白,朱伯阳,任晓明,等.羊接种布鲁氏菌S2菌苗抗体消长规律研究[J].中国农学通报,2018,34(9):119-123.
[13]申捷,宝音达来,马立峰,等.布鲁氏菌病疫苗(S2株)免疫羊抗体消长规律的研究[J].中国动物检疫,2015,32(1):68-73.
[14]于广秋.羊布氏杆菌病阴道免疫法推广[J].中国畜禽种业,2017,13(4):84-85.
[15]张雷,曹东,郑文金,等.阴道免疫对羊布鲁氏菌病疫苗抗体变化的影响[J].黑龙江畜牧兽医,2018,(2):115-117.
[2]陈丹,柳晓琳,刘孝刚,等.布鲁氏菌病流行趋势及其防治措施的研究进展[J].中国地方病防治杂志,2017,(1):26-28.
[3]Rahman C V,Kuhn G,White L J,et al.PLGA/PEG-hydro gel composite scaffolds with controllable mechanical properties[J].J Biomed Mater Res B Appl Biomater,2013,101(4):648.
[4]Zhong Xing,Wang Yu-Xin,Wang Shi-chang.Pressure dependence of the volume phase-transition of temperature-sensitive gels[J].Chen Engin Sci,1996,51(12):3235-3239.
[5]Alexandre M,Sun Hao,Ghaleb A H,et al.Drug delivery in pluronic micelles:effect of high-frequency ultrasound on drug release from micelles and intracellular uptake[J].J Con Rel,2002,84(1-2):39-47.
[6]石颖.喷鼻流感三价裂解疫苗的制备和免疫效果的初步研究[D].北京:中国人民解放军军事医学科学院,2012.
[7]朱晓波.球虫疫苗稀释剂的制备及其对疫苗免疫效果的影响[D].晋中:山西农业大学,2014.
[8]王霄旸,王米,张可煜,等.采用泊洛沙姆为佐剂的O型口蹄疫多肽疫苗对大鼠脾细胞的增值作用[J].中国动物传染病学报,2015,23(2):74-77.
[9]管晓燕,李敏,刘建国,等.温敏型生物降解水凝胶载体防龋基因疫苗p VAX1-spap/A经不同途径免疫新西兰大白兔的实验研究[J].口腔医学研究,2014,10:28.
[10]翟良,王霄旸,王昊欣,等.蛋白载体温敏凝胶制备及释放度研究[J].中国兽药杂志,2014,48(5):32-35.
[11]Juan E U,Susana L,Rodolfo A U.Evaluation of Brucella abortus phosphoglucomutase(pgm)mutant as a new live rough-phenotype vaccine[J].Infect Immun,2003,71:6264.
[12]李晓白,朱伯阳,任晓明,等.羊接种布鲁氏菌S2菌苗抗体消长规律研究[J].中国农学通报,2018,34(9):119-123.
[13]申捷,宝音达来,马立峰,等.布鲁氏菌病疫苗(S2株)免疫羊抗体消长规律的研究[J].中国动物检疫,2015,32(1):68-73.
[14]于广秋.羊布氏杆菌病阴道免疫法推广[J].中国畜禽种业,2017,13(4):84-85.
[15]张雷,曹东,郑文金,等.阴道免疫对羊布鲁氏菌病疫苗抗体变化的影响[J].黑龙江畜牧兽医,2018,(2):115-117.