壬二酸佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答效应分析
|
摘要
目的探讨壬二酸作为佐剂对甲型肝炎减毒活疫苗(HepA-1)诱导小鼠体液免疫应答的影响,以评价其佐剂效应。方法分别将四种不同剂量的壬二酸(0.5 mg,1 mg,2 mg,3 mg)与HepA-1共同免疫ICR小鼠分别作为佐剂组1、佐剂组2、佐剂组3、佐剂组4,同时设空白对照组、单纯抗原组、铝佐剂组,共免疫1次,于免疫后4周、8周、12周、16周进行尾静脉采血,采用间接酶联免疫吸附法(ELISA)检测小鼠血清抗甲型肝炎病毒(HAV)IgG抗体水平。结果除空白对照组外,各组小鼠免疫后4周、8周、12周、16周,均产生抗HAVIgG抗体,抗体水平随免疫时间的延长逐渐升高,于8、12周达到峰值。免疫后4周,佐剂组4(壬二酸3 mg)产生的抗甲型肝炎病毒(HAV)IgG抗体水平与铝佐剂组相比较差异有统计学意义(P<0.05,t=2.640)。同时,佐剂组4在免疫后4周、8周和12周产生的抗甲型肝炎病毒(HAV)IgG抗体水平较高,显著高于单纯抗原组(P<0.05,t值分别为3.134、2.751、2.743)。结论适当剂量的壬二酸可快速、显著提高HepA-1诱导小鼠的体液免疫应答,有望成为一种新型的人用疫苗佐剂。
Objective To investigate the effect of azelaic acid as an adjuvant on humoral immune response induced by hepatitis A virus(HAV) attenuated live vaccine(HepA-1) in mice to evaluate its adjuvant effect. Methods The ICR mice were injected respectively with HepA-1(18 EU) and Al(OH)_3(300 mg) and four different doses(0.5 mg, 1 mg, 2 mg, 3 mg) of azelaic acid as adjuvant group, 300 μL for each and one immunization. At the same time, set up the blank group, pure vaccine group, aluminum adjuvant control group. The sera from tail vein blood of mice were collected at the end of 4, 8, 12 and 16 weeks after immunization and the specific IgG antibody against HAV were detected by indirect ELISA method. Results In addition to the blank group, mice in various groups generated Anti-HAV IgG at 4,8,12,16 weeks after the immunization. The antibody levels increased as time went on and reached peak at the 8 th and 12 th week, then decreased gradually. At 4 weeks, the level of anti-hepatitis A virus(HAV)IgG produced by adjuvant group 4(azelaic acid 3 mg) was significantly higher than aluminum adjuvant control group(P<0.05, t = 2.640). At the same time, the level of anti-hepatitis A virus(HAV) IgG in adjuvant group 4 was significantly higher at 4 th week, 8 th week and 12 th week than pure antigen group(P<0.05, t = 3.134,2.751,2.743). Conclusion Appropriate dose of azelaic acid can obviously enhance the HepA-1 induced humoral immune response in mice. Azelaic acid is expected to become a new adjuvant for human vaccines.
Objective To investigate the effect of azelaic acid as an adjuvant on humoral immune response induced by hepatitis A virus(HAV) attenuated live vaccine(HepA-1) in mice to evaluate its adjuvant effect. Methods The ICR mice were injected respectively with HepA-1(18 EU) and Al(OH)_3(300 mg) and four different doses(0.5 mg, 1 mg, 2 mg, 3 mg) of azelaic acid as adjuvant group, 300 μL for each and one immunization. At the same time, set up the blank group, pure vaccine group, aluminum adjuvant control group. The sera from tail vein blood of mice were collected at the end of 4, 8, 12 and 16 weeks after immunization and the specific IgG antibody against HAV were detected by indirect ELISA method. Results In addition to the blank group, mice in various groups generated Anti-HAV IgG at 4,8,12,16 weeks after the immunization. The antibody levels increased as time went on and reached peak at the 8 th and 12 th week, then decreased gradually. At 4 weeks, the level of anti-hepatitis A virus(HAV)IgG produced by adjuvant group 4(azelaic acid 3 mg) was significantly higher than aluminum adjuvant control group(P<0.05, t = 2.640). At the same time, the level of anti-hepatitis A virus(HAV) IgG in adjuvant group 4 was significantly higher at 4 th week, 8 th week and 12 th week than pure antigen group(P<0.05, t = 3.134,2.751,2.743). Conclusion Appropriate dose of azelaic acid can obviously enhance the HepA-1 induced humoral immune response in mice. Azelaic acid is expected to become a new adjuvant for human vaccines.
引文
[1]Schijns VE,Lavelle C.Trends in vaccine adjuvants[J].Expert Rev Vaccines,2011,10(4):539-550.
[2]李学钊,洪炀,王治仓,等.免疫佐剂研究进展及前景展望[J].中国动物传染病学报,2014(4):81-86.
[3]何慧,李彦涵,李建芳,等.吲哚胺-2,3-双加氧酶抑制剂联合铝佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答的效应分析[J].实验动物与比较医学,2017,37(2):108-112.
[4]Monaci E,Mancini F,Lofano G,et al.MF59-and Al(OH)_3-adjuvanted Staphylococcus aureus(4C-staph)vaccines Induce sustained protective humoral and cellular immune responses,with a critical role for effector CD4 T cells at low antibody titers[J].Front Immunol,2015,6:439.
[5]Del Rosso JQ.Azelaic acid topical formulations:differentiation of 15%gel and 15%foam[J].J Clin Aesthet Dermatol,2017,10(3):37-40.
[6]Schulte BC,Wu W,Rosen T.Azelaic acid:evidence-based update on mechanism of action and clinical application[J].J Drugs Dermatol,2015,14(9):964-968.
[7]张宜,刘珍勇.壬二酸临床前药学研究进展[J].中国药业,2007,16(18):57-58.
[8]Mastrofrancesco A,Ottaviani M,Aspite N,et al.Azelaic acid modulates the inflammatory response in normal human keratinocytes through PPARγactivation[J].Exp Dermatol,2010,19(9):813-820.
[9]Draelos ZD,Elewski BE,Harper JC,et al.A phase 3randomized,double-blind,vehicle-controlled trial of azelaic acid foam 15%in the treatment of papulopustular rosacea[J].Cutis,2015,96(1):54-61.
[10]马佳.壬二酸(AZA)的抗菌及对黑素细胞与黑素瘤细胞的抑制作用[D].合肥:安徽医科大学,2006.
[11]Luo M,Shao B,Yu JY,et al.Simultaneous enhancement of cellular and humoral immunity by the high salt formulation of Al(OH)3 adjuvant[J].Cell Res,2017,27(4):586-589.
[12]王丽萍,王越,王海漩,等.酵母多糖-角鲨烯复合佐剂对甲型肝炎和乙型肝炎抗原诱导小鼠体液免疫应答的增强作用[J].中国生物制品学杂志,2013,26(4):473-476.
[13]马静,王海漩,李思瑾,等.4-苯基咪唑+氢氧化铝复合佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答的影响[J].实验动物与比较医学,2017,37(2):102-107.
[14]王玮,余柄廷,李建芳,等.低分子量肝素钠佐剂对甲型肝炎病毒疫苗诱导小鼠体液免疫应答的影响[J].中国比较医学杂志,2016,26(1):19-24.
[15]张周洋,耿兴超,汪巨峰,等.疫苗佐剂最新研究进展[J].中国新药杂志,2013(1):34-42.
[16]Sivakumar SM,Safhi MM,Kannadasan M,et al.Vaccine adjuvants-Current status and prospects on controlled release adjuvancity[J].Saudi Pharm J,2011,19(4):197-206.
[17]Aguilar JC,Rodríguez EG.Vaccine adjuvants revisited[J].Vaccine,2007,25(19):3752-3762.
[18]Graupe K,Cunliffe WJ,Gollnick HP,et al.Efficacy and safety of topical azelaic acid(20 percent cream):an overview of results from European clinical trials and experimental reports[J].Cutis,1996,57(1 Suppl):20-35.
[19]Worret WI,Fluhr JW.Acne therapy with topical benzoyl peroxide,antibiotics and azelaic acid[J].J Dtsch Dermatol Ges,2006,4(4):293-300.
[20]Gollnick HP,Graupe K,Zaumseil RP.Comparison of combined azelaic acid cream plus oral minocycline with oral isotretinoin in severe acne[J].Eur J Dermatol,2001,11(6):538-544.
[2]李学钊,洪炀,王治仓,等.免疫佐剂研究进展及前景展望[J].中国动物传染病学报,2014(4):81-86.
[3]何慧,李彦涵,李建芳,等.吲哚胺-2,3-双加氧酶抑制剂联合铝佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答的效应分析[J].实验动物与比较医学,2017,37(2):108-112.
[4]Monaci E,Mancini F,Lofano G,et al.MF59-and Al(OH)_3-adjuvanted Staphylococcus aureus(4C-staph)vaccines Induce sustained protective humoral and cellular immune responses,with a critical role for effector CD4 T cells at low antibody titers[J].Front Immunol,2015,6:439.
[5]Del Rosso JQ.Azelaic acid topical formulations:differentiation of 15%gel and 15%foam[J].J Clin Aesthet Dermatol,2017,10(3):37-40.
[6]Schulte BC,Wu W,Rosen T.Azelaic acid:evidence-based update on mechanism of action and clinical application[J].J Drugs Dermatol,2015,14(9):964-968.
[7]张宜,刘珍勇.壬二酸临床前药学研究进展[J].中国药业,2007,16(18):57-58.
[8]Mastrofrancesco A,Ottaviani M,Aspite N,et al.Azelaic acid modulates the inflammatory response in normal human keratinocytes through PPARγactivation[J].Exp Dermatol,2010,19(9):813-820.
[9]Draelos ZD,Elewski BE,Harper JC,et al.A phase 3randomized,double-blind,vehicle-controlled trial of azelaic acid foam 15%in the treatment of papulopustular rosacea[J].Cutis,2015,96(1):54-61.
[10]马佳.壬二酸(AZA)的抗菌及对黑素细胞与黑素瘤细胞的抑制作用[D].合肥:安徽医科大学,2006.
[11]Luo M,Shao B,Yu JY,et al.Simultaneous enhancement of cellular and humoral immunity by the high salt formulation of Al(OH)3 adjuvant[J].Cell Res,2017,27(4):586-589.
[12]王丽萍,王越,王海漩,等.酵母多糖-角鲨烯复合佐剂对甲型肝炎和乙型肝炎抗原诱导小鼠体液免疫应答的增强作用[J].中国生物制品学杂志,2013,26(4):473-476.
[13]马静,王海漩,李思瑾,等.4-苯基咪唑+氢氧化铝复合佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答的影响[J].实验动物与比较医学,2017,37(2):102-107.
[14]王玮,余柄廷,李建芳,等.低分子量肝素钠佐剂对甲型肝炎病毒疫苗诱导小鼠体液免疫应答的影响[J].中国比较医学杂志,2016,26(1):19-24.
[15]张周洋,耿兴超,汪巨峰,等.疫苗佐剂最新研究进展[J].中国新药杂志,2013(1):34-42.
[16]Sivakumar SM,Safhi MM,Kannadasan M,et al.Vaccine adjuvants-Current status and prospects on controlled release adjuvancity[J].Saudi Pharm J,2011,19(4):197-206.
[17]Aguilar JC,Rodríguez EG.Vaccine adjuvants revisited[J].Vaccine,2007,25(19):3752-3762.
[18]Graupe K,Cunliffe WJ,Gollnick HP,et al.Efficacy and safety of topical azelaic acid(20 percent cream):an overview of results from European clinical trials and experimental reports[J].Cutis,1996,57(1 Suppl):20-35.
[19]Worret WI,Fluhr JW.Acne therapy with topical benzoyl peroxide,antibiotics and azelaic acid[J].J Dtsch Dermatol Ges,2006,4(4):293-300.
[20]Gollnick HP,Graupe K,Zaumseil RP.Comparison of combined azelaic acid cream plus oral minocycline with oral isotretinoin in severe acne[J].Eur J Dermatol,2001,11(6):538-544.