Evaluation of the protections of multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene

ZHANG Qiao; et al

doi:10.13406/j.cnki.cyxb.2010.05.009

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Evaluation of the protections of multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene

更新时间：2024-01-17

- Evaluation of the protections of multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene
- Journal of Chongqing Medical University Vol. 35, Issue 5, Pages: 711-714(2010)
- 作者机构：
  
  第三军医大学新桥医院全军呼吸内科研究所
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.05.009
  CLC： R392
- Published：2010，
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[1]张巧,林科雄,马千里,王长征.肺炎链球菌溶菌酶作为肺炎链球菌多价核酸疫苗候选抗原的保护效能评价[J].重庆医科大学学报,2010,35(05):711-714.

ZHANG Qiao, et al. Evaluation of the protections of multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene. [J]. Journal of Chongqing Medical University 35(5):711-714(2010)
[1]张巧,林科雄,马千里,王长征.肺炎链球菌溶菌酶作为肺炎链球菌多价核酸疫苗候选抗原的保护效能评价[J].重庆医科大学学报,2010,35(05):711-714. DOI： 10.13406/j.cnki.cyxb.2010.05.009.

ZHANG Qiao, et al. Evaluation of the protections of multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene. [J]. Journal of Chongqing Medical University 35(5):711-714(2010) DOI： 10.13406/j.cnki.cyxb.2010.05.009.

摘要

目的:肺炎链球菌溶菌酶（N-acetylmuramoyl-L-alanine amidase

LytA）作为肺炎链球菌多价核酸疫苗候选抗原的保护效能评价。方法:PCR技术从肺炎链球菌R6标准株扩增LytA基因序列

构建重组真核表达质粒pcDNA3.1-LytA

免疫印迹（western-blot）检测重组质粒在哺乳动物BHK-21细胞中的表达。与前期构建的肺炎链球菌表面蛋白A（Pneumococcal surface protein A

PspA）N端序列基因构建的质粒pcDNA3.1-PspA’核酸疫苗联合或单独肌肉注射免疫BABL/c小鼠

检测各组小鼠血清特异性抗体IgG水平

并观察免疫小鼠肺炎链球菌D39腹腔攻击21d生存情况。结果:ELISA检测发现3组（pcDNA3.1-LytA组

pcDNA3.1-PspA’组

pcDNA3.1-LytA+pcDNA3.1-PspA’组）实验组小鼠特异性IgG水平较空质粒对照组显著升高（P<0.001）

pcDNA3.1-LytA+pcDNA3.1-PspA’组小鼠抗LytA特异性抗体IgG水平较pcDNA3.1-LytA组明显升高（P<0.01）

但是与pcDNA3.1-PspA’组比较抗PspA特异性抗体IgG水平无显著差异（P>0.05）。小鼠攻击试验提示pcDNA3.1-LytA+pcDNA3.1-PspA’组小鼠中位生存时间较pcDNA3.1-LytA组及空质粒对照组显著延长（P<0.01）

但与pcDNA3.1-PspA’组比较中位生存时间无显著差异（P>0.05）

生存曲线相似。结论:LytA联合PspA’的肺炎链球菌多价核酸疫苗免疫效能并不优于单一抗原PspA’的肺炎链球菌核酸疫苗

LytA作为肺炎链球菌多价核酸疫苗的优势候选抗原组分之一有待进一步研究。

Abstract

Objective:To evaluate the ability of the cooperative protection of the Strep to coccus pneumoniae multivalent DNA vaccine containing lytA gene(N-acetylmuramoyl-L-alanine amidase

LytA)and pspA’gene(N fragment of pneumococcal surface protein A

PspA)against intraperitoneal Streptococcus pneumoniae D39 infection.Methods The genomic DNA of the standard Streptococcus pneumoniae strain D39 was isolated.LytA gene fragment was amplified by polymerase chain reaction(PCR)and cloned into the eukaryotic expression vector pcDNA3.1(+)

which was transfected into BHK-21 cells using Lipofectamine TM2000

and then the expression of LytA protein was detected with Western-blot.Furthermore

three formats of recombinant plasmid DNA vaccines(pcDNA3.1-LytA+pcDNA3.1-PspA’

pcDNA3.1-PspA’

pcDNA3.1-LytA)were used to inject intramuscularly BALB/c mice

with pcDNA3.1(+)plasmid and PBS as controls.The levels of antibodies against PspA’and LytA proteins were checked.Live times of intraperitoneal infection mice by Streptococcus pneumoniae D39 in BALB/c models were analysis.Results:The three DNA vaccines all producted high levels serum special IgG antibodies.The anti-LytA antibodies level elicited by the DNA vaccine pcDNA3.1-LytA + pcDNA3.1-PspA’was higher than the DNA vaccine pcDNA3.1-LytA(P<0.01)

but the anti-PspA’antibodies level elicited by the DNA vaccine pcDNA3.1-LytA + pcDNA3.1-PspA’had no significant statistics deviation(P>0.05).The result of the effect on intraperitoneal infection by Streptococcus pneumoniae D39 in BALB/c mice models showed the median live times in pcDNA3.1-LytA + pcDNA3.1-PspA’group was longer than pcDNA3.1-LytA group and the control groups(P<0.01)

but no significant statistics deviation was found between pcDNA3.1-LytA + pcDNA3.1-PspA’group with pcDNA3.1-PspA’group(P >0.05).Conclusion:The multivalent DNA vaccine of Streptococcus pneumoniae containing LytA gene and pspA gene had no the higher performance protection than the DNA vaccine only containing pspA gene against Streptococcus pneumoniae.It is should be further evaluated that LytA gene is concluded as the one of advantages candidate antigens of the Streptococcus pneumoniae multivalent DNA vaccine.

关键词

肺炎链球菌肺炎链球菌溶菌酶肺炎链球菌表面蛋白A多价疫苗

Keywords

Streptococcus pneumoniaeLytAPspAMultivalent vaccine

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