中国眼耳鼻喉科杂志
中國眼耳鼻喉科雜誌
중국안이비후과잡지
CHINESE JOURNAL OF OPHTHALMOLOGY AND OTOLARYNGOLOGY
2009年
4期
212-214,附1
,共4页
徐江红%王正敏%陈兵%章德广%戴文佳%朱美美%徐晨媚
徐江紅%王正敏%陳兵%章德廣%戴文佳%硃美美%徐晨媚
서강홍%왕정민%진병%장덕엄%대문가%주미미%서신미
中耳炎%链球菌,肺炎%疫苗%DNA
中耳炎%鏈毬菌,肺炎%疫苗%DNA
중이염%련구균,폐염%역묘%DNA
Otitis media%Streptococcus pneumoniae%Vaccines%DNA
目的 构建肺炎链球菌自溶素(pneumococcal autolysin,LytA)核酸疫苗,并研究其免疫原性.方法 采用聚合酶链反应方法从肺炎链球菌基因组中克隆肺炎链球菌自溶素基因,然后将该基因插入到pVAX1真核表达载体中,构建表达LytA的重组质粒,制备成超螺旋比例大于90%的核酸疫苗.实验组:BALB/c小鼠5只,对其分3次肌内注射核酸疫苗(100 μg);对照组:BALB/c小鼠5只,肌内注射pVAXI空载体(100μg)3次.分别采集小鼠血清,采用间接酶联免疫吸附方法测定血清LytA特异性抗体水平.结果 扩增出的LytA基因(957 bp)与基因库中LytA的编码序列(957 bp)一致.成功构建了LytA核酸疫苗.实验组小鼠的血清Ly-tA抗体水平明显高于对照组,差异有统计学意义(P<0.01).结论 成功克隆了LytA基因,制备了LytA核酸疫苗,后者可在小鼠体内诱导较强的特异性体液免疫反应.
目的 構建肺炎鏈毬菌自溶素(pneumococcal autolysin,LytA)覈痠疫苗,併研究其免疫原性.方法 採用聚閤酶鏈反應方法從肺炎鏈毬菌基因組中剋隆肺炎鏈毬菌自溶素基因,然後將該基因插入到pVAX1真覈錶達載體中,構建錶達LytA的重組質粒,製備成超螺鏇比例大于90%的覈痠疫苗.實驗組:BALB/c小鼠5隻,對其分3次肌內註射覈痠疫苗(100 μg);對照組:BALB/c小鼠5隻,肌內註射pVAXI空載體(100μg)3次.分彆採集小鼠血清,採用間接酶聯免疫吸附方法測定血清LytA特異性抗體水平.結果 擴增齣的LytA基因(957 bp)與基因庫中LytA的編碼序列(957 bp)一緻.成功構建瞭LytA覈痠疫苗.實驗組小鼠的血清Ly-tA抗體水平明顯高于對照組,差異有統計學意義(P<0.01).結論 成功剋隆瞭LytA基因,製備瞭LytA覈痠疫苗,後者可在小鼠體內誘導較彊的特異性體液免疫反應.
목적 구건폐염련구균자용소(pneumococcal autolysin,LytA)핵산역묘,병연구기면역원성.방법 채용취합매련반응방법종폐염련구균기인조중극륭폐염련구균자용소기인,연후장해기인삽입도pVAX1진핵표체재체중,구건표체LytA적중조질립,제비성초라선비례대우90%적핵산역묘.실험조:BALB/c소서5지,대기분3차기내주사핵산역묘(100 μg);대조조:BALB/c소서5지,기내주사pVAXI공재체(100μg)3차.분별채집소서혈청,채용간접매련면역흡부방법측정혈청LytA특이성항체수평.결과 확증출적LytA기인(957 bp)여기인고중LytA적편마서렬(957 bp)일치.성공구건료LytA핵산역묘.실험조소서적혈청Ly-tA항체수평명현고우대조조,차이유통계학의의(P<0.01).결론 성공극륭료LytA기인,제비료LytA핵산역묘,후자가재소서체내유도교강적특이성체액면역반응.
Objective To construct pneumacoccal autolysin(LytA) DNA vaccine and to study its immunogenicity. Methods The LytA gene was amplified from the pneumococeal genome by polymerase chain reaction and then was inserted into the enkaryotic expression vector pVAX1 to construct the recombinant expression plasmid. The BALB/cmice were intramuscularly injected with LytA DNA vaccine(100 μg) in the experimental group(n = 5) or with pVAX1 (100 μg) in the control group(n =5) for 3 times respectively. The LytA-specific antibody level was measured by the enzyme-linked immunosorbent assay method. Results The DNA sequence of the cloned LytA (957 bp) was consistent with the GenBank data (957 bp) and the recombinant expression plasmid pVAX1-LytA was successfully constructed. The LytA-specific antibody level was significantly higher in the experimental group than in the control group (P < 0.01). Conclusions The LytA gene was successfully cloned. LytA DNA vaccine was successfully prepared which could induce powerful humoral immune reaction in mice. (Chin J Ophthalmol and Otorhinolaryngol,2009,9:212-214)
