呼吸道感染细菌联合疫苗体内实验研究
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摘要
目的通过小鼠体内实验进行免疫学指标检测,观察其对机体免疫功能的影响,为研制我国呼吸道感染细菌联合疫苗提供理论依据。方法1分离培养我国小儿呼吸道感染常见6种细菌,按洗涤、超声波破碎、沉淀、透析、分光光度计测定等过程制备细菌蛋白,等量混合配成一定浓度作为实验用联合疫苗。2取无特定病原体级小鼠40只,随机分成4组,每组10只。全部实验小鼠饲喂基础日粮,适应性喂养10d后,实验组分别用浓度为0.1、0.2、0.4g/L联合疫苗免疫,每天上午灌服0.2mL。对照组每天灌服0.2mL生理盐水。免疫10d。处死小鼠,分别测定联合疫苗组小鼠巨噬细胞吞噬功能、细胞免疫功能、血清中溶血素含量以及胸腺指数和脾脏指数的影响。结果联合疫苗组的巨噬细胞吞噬率和吞噬指数、左右后足质量差、溶血素吸光度值、脾脏指数和胸腺指数均显著高于对照组(P<0.05),在实验设计疫苗浓度范围各组中,不同组间检测值随疫苗浓度的增加而加大,且各实验组间也均存在明显差异(P<0.05)。结论制备的联合疫苗可提高机体免疫功能,不仅能提高细胞免疫功能,对体液免疫功能也具有明显影响,在实验浓度范围内,疫苗浓度越高对机体免疫功能影响越明显。
Objective The immune function of the mice was detected through the in vivo experiment in mice,which will provide theoretical basis for the development of the bacterial combined vaccine of respiratory tract infection in our country.Methods 1Six common bacterias in children with respiratory tract infection in our country were isolated and cultured.Bacterial proteins were prepared by washing, ultrasonic crushing, precipitation, dialysis and spectrophotometer,and the equivalent amount of the mixture was used as the experimental combined vaccine.2Forty specific pathogen free mice were randomly divided into four groups,with 10 rats in each group.All mice were fed with a basal diet.After adaptive feeding for 10 days,the experimental group were fed with concentrations of 0.1,0.2,0.4g/L combined vaccine,with daily morning irrigation of 0.2 mL.The mice in normal control group were orally administered daily with physiological saline 0.2 mL.All groups were immuned for 10 days.Effects of combined vaccine on the phagocytic function of macrophage,the cellular immune function of mice,the changes of serum hemolysin content and the effect of thymus index and spleen index determinated.Results Compared with the normal control group,the macrophage phagocytic rate and phagocytic index,left and right foot weight,the absorbance value of serum hemolysin,the spleen index and thymus index of combined vaccine groups were significantly increased(P<0.05).In the experimental design of vaccine concentration range,the detection values in different groups were increased with the increasement of vaccine concentration,and there were significant differences among the test groups(P<0.05).Conclusion Combined vaccine can improve the immune function of organism.Combined vaccine can not only improve the cellular immune function,but also has obvious effect on humoral immune function.In the experimental concentration range,the higher the vaccine concentration,the more obvious effect on the immune function of the body.
Objective The immune function of the mice was detected through the in vivo experiment in mice,which will provide theoretical basis for the development of the bacterial combined vaccine of respiratory tract infection in our country.Methods 1Six common bacterias in children with respiratory tract infection in our country were isolated and cultured.Bacterial proteins were prepared by washing, ultrasonic crushing, precipitation, dialysis and spectrophotometer,and the equivalent amount of the mixture was used as the experimental combined vaccine.2Forty specific pathogen free mice were randomly divided into four groups,with 10 rats in each group.All mice were fed with a basal diet.After adaptive feeding for 10 days,the experimental group were fed with concentrations of 0.1,0.2,0.4g/L combined vaccine,with daily morning irrigation of 0.2 mL.The mice in normal control group were orally administered daily with physiological saline 0.2 mL.All groups were immuned for 10 days.Effects of combined vaccine on the phagocytic function of macrophage,the cellular immune function of mice,the changes of serum hemolysin content and the effect of thymus index and spleen index determinated.Results Compared with the normal control group,the macrophage phagocytic rate and phagocytic index,left and right foot weight,the absorbance value of serum hemolysin,the spleen index and thymus index of combined vaccine groups were significantly increased(P<0.05).In the experimental design of vaccine concentration range,the detection values in different groups were increased with the increasement of vaccine concentration,and there were significant differences among the test groups(P<0.05).Conclusion Combined vaccine can improve the immune function of organism.Combined vaccine can not only improve the cellular immune function,but also has obvious effect on humoral immune function.In the experimental concentration range,the higher the vaccine concentration,the more obvious effect on the immune function of the body.
引文
[1]张敬,朱丽华,张庆波.呼吸道感染细菌联合疫苗体外实验研究[J].实用预防医学,2015,22(6):746-748.
[2]朱颋,陈颖张.匹多莫德佐治小儿反复呼吸道感染疗效观察[J].临床合理用药,2015,8(11A):81-82.
[3]袁婷,关晓菲,马翠卿,等.A族链球菌M蛋白相关疫苗的研究进展[J].河北医科大学学报,2015,36(1):103-107.
[4]徐宗凯,林青青,周梦莹,等.三种李斯特菌菌体蛋白提取方法的比较[J].生命科学研究,2015,19(1):29-33.
[5]孙侠,韩长城,牛玉杰,等.溴氰菊酯对小鼠免疫系统的损伤作用[J].河北医科大学学报,2005,26(1):1-4.
[6]蔡岩,赵晓青,李继宏,等.乙型肝炎卡介苗联合疫苗免疫豚鼠及小鼠的病理学研究[J].微生物学杂志,2012,32(6):82-86.
[7]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[8]Cazzola M,Anapurapu S,Page CP.Polyvalent mechanical bacterial lysate for the prevention of recurrent respiratory infections:a meta-analysis[J].Pulm Pharmacol Ther,2012,25:62-68.
[9]洪建国.儿童反复呼吸道感染细菌提取物的临床应用[J].中国实用儿科杂志,2013,28(3):165-168.
[10]Rossi ME,Legal L,Azzari C,et al.Effect of a polyvalent bacterial preparation on nature kill cell activity in children with recurrent respiratory infections[J].Int J Imumunotherapy,1993,9(4):225-233.
[11]赵志强,贾丽霞,张庆波,等.黄酮对免疫功能影响的体内研究[J].临床荟萃,2014,29(1):49-51.
[12]张伟.泛福舒辅助治疗儿童支气管哮喘的临床观察[J].中国医药指南,2015,13(13):70.
[13]吴永利,孔令茹,张洁,等.泛福舒治疗小儿反复呼吸道感染疗效观察[J].临床荟萃,2008,23(9):659-660.
[14]罗新舟,孙江桥.复方黄芪合剂对小鼠免疫功能影响[J].辽宁中医药大学学报,2014,16(1):21-22.
[15]王秀青,孙慧敏,朱明,等.CecropinA-thanatin杂合肽对小鼠生长性能及免疫功能的影响[J].饲料研究,2015,8:65-70.
[16]刘印华,赵志强,李树义,等.黄芪多糖对免疫功能影响的体内实验研究[J].河北医药,2015,37(4):485-487.
[17]宋媛媛,周越,孙守兵,等.丁香苷对免疫功能低下小鼠的免疫调节作用[J].中药药理与临床,2013,29(2):44-47.
[18]Hamada K,Sakaue M,Sarkar A,et al.Immune responses to repetitive adenovirus-mediated gene transfer and restoration of gene expression by cyclophosphamide or etoposide[J].Gynecol Oncol,2005,99(3):177-186.
[2]朱颋,陈颖张.匹多莫德佐治小儿反复呼吸道感染疗效观察[J].临床合理用药,2015,8(11A):81-82.
[3]袁婷,关晓菲,马翠卿,等.A族链球菌M蛋白相关疫苗的研究进展[J].河北医科大学学报,2015,36(1):103-107.
[4]徐宗凯,林青青,周梦莹,等.三种李斯特菌菌体蛋白提取方法的比较[J].生命科学研究,2015,19(1):29-33.
[5]孙侠,韩长城,牛玉杰,等.溴氰菊酯对小鼠免疫系统的损伤作用[J].河北医科大学学报,2005,26(1):1-4.
[6]蔡岩,赵晓青,李继宏,等.乙型肝炎卡介苗联合疫苗免疫豚鼠及小鼠的病理学研究[J].微生物学杂志,2012,32(6):82-86.
[7]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[8]Cazzola M,Anapurapu S,Page CP.Polyvalent mechanical bacterial lysate for the prevention of recurrent respiratory infections:a meta-analysis[J].Pulm Pharmacol Ther,2012,25:62-68.
[9]洪建国.儿童反复呼吸道感染细菌提取物的临床应用[J].中国实用儿科杂志,2013,28(3):165-168.
[10]Rossi ME,Legal L,Azzari C,et al.Effect of a polyvalent bacterial preparation on nature kill cell activity in children with recurrent respiratory infections[J].Int J Imumunotherapy,1993,9(4):225-233.
[11]赵志强,贾丽霞,张庆波,等.黄酮对免疫功能影响的体内研究[J].临床荟萃,2014,29(1):49-51.
[12]张伟.泛福舒辅助治疗儿童支气管哮喘的临床观察[J].中国医药指南,2015,13(13):70.
[13]吴永利,孔令茹,张洁,等.泛福舒治疗小儿反复呼吸道感染疗效观察[J].临床荟萃,2008,23(9):659-660.
[14]罗新舟,孙江桥.复方黄芪合剂对小鼠免疫功能影响[J].辽宁中医药大学学报,2014,16(1):21-22.
[15]王秀青,孙慧敏,朱明,等.CecropinA-thanatin杂合肽对小鼠生长性能及免疫功能的影响[J].饲料研究,2015,8:65-70.
[16]刘印华,赵志强,李树义,等.黄芪多糖对免疫功能影响的体内实验研究[J].河北医药,2015,37(4):485-487.
[17]宋媛媛,周越,孙守兵,等.丁香苷对免疫功能低下小鼠的免疫调节作用[J].中药药理与临床,2013,29(2):44-47.
[18]Hamada K,Sakaue M,Sarkar A,et al.Immune responses to repetitive adenovirus-mediated gene transfer and restoration of gene expression by cyclophosphamide or etoposide[J].Gynecol Oncol,2005,99(3):177-186.