脂多糖模拟肽13L诱导对小鼠内毒素休克的保护反应
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国家自然科学基金资助项目(30471550).


Protective Immunity Induced by Peptide Mimics to LPS in Mice With Endotoxic Shock
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This work was supported by a grant from The National Natural Sciences Foundation of China(30471550).

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    摘要:

    为研究LPS 2630表位模拟肽13L是否能诱导抗脂多糖(LPS)抗体的产生、对细菌攻击及内毒素休克的保护性反应,合成了模拟LPS表位的短肽13L.用合成肽13L-蓝载体(blue carrier, BC)交联物免疫Balb/C小鼠,观察小鼠体内抗LPS抗体产生的规律,并观察免疫小鼠细菌感染存活时间,用颈总动脉插管测定技术观察免疫小鼠内毒素休克的血压变化.结果显示,合成肽13L-BSA交联物能与兔抗鼠伤寒和大肠杆菌LPS抗血清及抗鼠伤寒LPS单抗结合,证明短肽13L可模拟LPS的抗原性.用13L-BC交联物免疫可诱发小鼠体内针对大肠杆菌LPS 2630和鼠伤寒沙门氏菌LPS 7261的抗体反应,该反应可被灭活大肠杆菌及两种LPS所加强,其抗体亚类主要为IgG2a,其次为IgG2b与IgM,而IgG1与IgG3含量极低.免疫13L-BC小鼠显示对细菌攻击的抵抗,与免疫BC对照小鼠比较,其存活时间分别为(12±1.3)天和(5.3±0.4)天(P < 0.01).免疫13L-BC小鼠对静脉注射LPS诱发内毒素休克的保护作用体现在血压下降幅度明显低于对照动物,在LPS 2630攻击后1、2、3及4 h时两组动物血压明显差别(P < 0.05、P < 0.01、 P < 0.05及 P < 0.01),而LPS 7261攻击后1、2、3及4 h时两组动物血压差别则略小于LPS 2630组(P > 0.05、P < 0.05、 P < 0.05及 P < 0.01).上述结果证明,LPS表位模拟肽13L交联物免疫小鼠可产生针对细菌攻击及内毒素休克的保护性效应,提示该模拟肽作为LPS交叉保护性疫苗候选的可行性.

    Abstract:

    To characterize a mimotop, peptide mimic to epitope on lipopolysaccharide (LPS) which was named as 13L and was expected to induce humoral immune response to LPS and a protective immunity , peptide 13L was synthesized and conjugated to Blue Carrier (BC) as immunogen. Balb/c mice were immunized with peptide 13L - BC conjugate and BC only as control. Anti-LPS antibodies of mice immunized with peptide 13L-BC were detected by ELISA using peptide 13L-BSA as coating antigen. Survival time of mice challenged with S. typhi, and dynamics of MAP/mmHg in mice injected with LPS were observed. The results showed that specific antibodies against S. typhi-LPS 7261 and E. coli-LPS 2630 were elicited in mice immunized with peptide 13L-BC conjugate, and could be boosted by inactive bacterial and LPS. The main isotype of anti-LPS antibodies were IgG2a, IgG2b and IgM, but less IgG1 and IgG3, The survival time of mice infected with S. typhi were (12±1.3) days and (5.3± 0.4) days(P < 0.01) in group immunized with peptide 13L conjugate and with BC, respectively. And the peptide 13L-BC can also elicit protective immunity against endotoxic shock by LPS. The dynamics of MAP/mmHg observed by carotid Artery catheterization showed a significant difference between mice immunized with peptide 13L-BC and mice immunized with BC only in 1, 2,3 and 4 hours after injection of LPS 2630(P < 0.05, P < 0.01, P < 0.05 and P < 0.01), and injection of LPS 7261(P > 0.05, P < 0.05, P < 0.05 and P < 0.01). These results suggested that the peptide 13L can mimic the antigenicity of LPS epitopes to induce secondary antibody response like as thymus-dependent antigen, and also can elicit a protective immunity of mice from infection with G- bacteria and endotoxic shock. This peptide mimics could be a new vaccine candidate of LPS.

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罗海波,郑健,朱平,黄来强,富宁.脂多糖模拟肽13L诱导对小鼠内毒素休克的保护反应[J].生物化学与生物物理进展,2008,35(11):1312-1319

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  • 收稿日期:2008-03-20
  • 最后修改日期:2008-06-21
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  • 在线发布日期: 2008-07-16
  • 出版日期: 2008-11-20