弓形体SAG1基因DNA诱导小鼠体液免疫应答及保护性研究

Study on the humoral and protective immunity responses induced by the recombinant plasmid with the SAG1 gene of toxoplasma godii in mice

目的 :用重组真核表达质粒 pc DNA3- SAG1直接免疫 BAL B/ c小鼠 ,观察其 DNA免疫所诱导的小鼠体液免疫反应和保护性作用。方法 :大量制备重组质粒 pc DNA3- SAG1,然后将其导入 BAL B/ c小鼠体内。抗体滴度用 EL ISA法进行测定 :采用 PCR方法对小鼠的血液组织外源基因进行检测 ;对试验组及对照组进行 RH株攻击感染 ,并对其进行分析。结果 :用 EL ISA法检测的抗体 Ig G滴度为 1∶ 2 5 6 0 ;免疫后三周、六个月从免疫鼠的血液组织中用 PCR仍可检测到特异的SAG1基因的存在 :体外弓形虫攻击感染实验组和对照组 ,可见实验组的存活时间较对照组时间延长 (P<0 .0 5 )。结论 :重组质粒 pc DNA3- SAG1免疫 BAL B/

Objective:Directly immunize mice with the recombinant plasmid pcDNA 3-SAG1 of Toxoplasma godii,to observe and test the humoral immune responses and protective induced by pcDNA 3-SAG1.Methods:Large scale prepared recombinant plasmid pcDNA 3-SAG1 was injected into the BALB/c mice by muscles.The IgG antibodies in serum from vaccinated mice were detected by enzyme-linked immunosorbent assy(ELISE).The SAG1 gene was detected in the blood of immunized mice detected by PCR metheod 6 month after last injection.The vaccinated mice were challenged by RH tachyzoit of Toxoplasma gondii.Result:The titer of specific IgG antibody in serum from vaccinated mice have been showed at 1∶2560.The SAG1 gene was detected in the blood of immunized mice by PCR at 3 week and 6 month after last injection.The survival time of immunized mice was longer than than of control group ( P <0.05) after being challenged with Toxoplasma gondii.Conclusion:DNA vaccine with the recombinant plasmid pcDNA 3-SAG1 could elicit BALB/c mice to generate humoral and protective immune responses for Toxoplasma godii infection.