论文部分内容阅读
目的制备氢氧化锌[Zn(OH)2]胶体,并检测其单独或与三磷酸腺苷二钠(Disodium adenosine tripho-sphate,简称ATP)联用对狂犬病疫苗诱导小鼠体液免疫应答的增强作用。方法采用葡萄糖酸锌作为锌源,与NaOH反应制备Zn(OH)2胶体溶液,对其进行纯化及检测,并进行小鼠急性毒性试验;将不同剂量的Zn(OH)2胶体单独或与不同剂量的ATP联用,与狂犬病疫苗(0.25 IU)混合,免疫ICR小鼠,并设生理盐水对照组和抗原对照组,每组8只,Zn(OH)2和Zn(OH)2+ATP和生理盐水对照组采用单次免疫,抗原对照组分别免疫1、2、3、5次,分别于初次免疫后4、8、12、16周采血,分离血清,检测血清中抗狂犬病病毒特异性IgG抗体水平。结果所制备Zn(OH)2胶体溶液最大吸收波长为213 nm,可观察到明显的丁达尔现象,浓度为6 mg/ml,经光学色差显微镜及扫描电镜观察合格,小鼠急性毒性试验未观察到急性毒副作用。大部分佐剂组抗体水平高于抗原单次免疫组,但均未达到抗原5次免疫组水平;0.27和0.21 mg Zn(OH)2组在初次免疫后8周内有高水平抗体产生;0.27 mg Zn(OH)2组在初次免疫后16周出现抗体水平反弹性升高;0.18 mg Zn(OH)2+1.0 mg ATP组和0.18 mg Zn(OH)2+0.5 mg ATP组在初次免疫后4周内产生的抗体水平较高,之后逐渐下降;2次和3次免疫抗原对照组在初次免疫后8周内,其抗体水平不及佐剂组。结论所制备的Zn(OH)2胶体佐剂单独或与ATP联用均可增强狂犬病疫苗诱导小鼠特异性体液免疫应答,且安全性良好,有望应用于疫苗生产。
OBJECTIVE To prepare zinc hydroxide [Zn (OH) 2] colloid and test its enhancing effect on humoral immune response induced by rabies vaccine alone or in combination with Disodium adenosine tripho-sphate (ATP). Methods Zn (OH) 2 colloidal solution was prepared by the reaction of zinc gluconate and zinc. The purified Zn (OH) 2 colloidal solution was tested and the acute toxicity test was performed in mice. Different doses of Zn (OH) Dose of ATP combined with rabies vaccine (0.25 IU), immunized ICR mice, and saline control group and antigen control group, with 8 rats in each group, Zn (OH) 2 and Zn (OH) 2 + ATP and The control group received single immunization and antigen control groups were immunized 1, 2, 3, 5 times respectively. Blood was collected at 4, 8, 12 and 16 weeks after primary immunization, respectively. Serum anti-rabies virus-specific IgG Antibody level. Results The maximum absorption wavelength of Zn (OH) 2 colloidal solution was 213 nm, and the obvious Tyndall phenomenon was observed at a concentration of 6 mg / ml. The results of optical color difference microscopy and scanning electron microscopy showed that the acute toxicity test was not observed To acute side effects. Most of the adjuvant group antibody levels were higher than the antigen single immunization group, but did not reach the level of five immunized groups antigen; 0.27 and 0.21 mg Zn (OH) 2 group within 8 weeks after the first immunization with high levels of antibody production; 0.27 Antibody level rebounded in the mg Zn (OH) 2 group at 16 weeks after the first immunization. In the group of 0.18 mg Zn (OH) 2 + 1.0 mg ATP and 0.18 mg Zn (OH) 2 + 0.5 mg ATP, The level of antibody produced in 4 weeks was higher, then gradually decreased. In the 2nd and 3rd immunization groups, the antibody level in the 2nd and 3rd immunization groups was less than that in the adjuvant group within 8 weeks after the first immunization. Conclusions The prepared Zn (OH) 2 colloid adjuvant can enhance the specific humoral immune response induced by rabies vaccine alone or in combination with ATP, and is safe and is expected to be used in vaccine production.