空肠弯曲菌PEB1的B细胞免疫优势表位的鉴定及保护效果的评价

冯健; 李运明; 刘毓刚; 王艳艳; 胡宗海; 熊杰; 张睿

doi:10.16016/j.1000-5404.201809180

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空肠弯曲菌PEB1的B细胞免疫优势表位的鉴定及保护效果的评价

基础医学 | 更新时间：2026-01-09

- 空肠弯曲菌PEB1的B细胞免疫优势表位的鉴定及保护效果的评价
- Fine-mapping of immunodominant linear B-cell epitopes of C.jejuni PEB1 antigen using short overlapping peptides
- 第三军医大学学报 2019年41卷第4期页码：302-307
- 作者机构：
  
  1.610083 成都，西部战区总医院：老年医学科
  2.610083 成都，西部战区总医院：信息科
  3.610083 成都，西部战区总医院：检验科
- 作者简介：
  
  [ 张睿，E-mail:echorayray@qq.com ZHANG Rui, E-mail: echorayray@qq.com
- 基金信息：
  
  国家自然科学基金青年科学基金(31400128);全军医学科技青年培育计划项目(孵化项目19QNP043)
- DOI：10.16016/j.1000-5404.201809180
  中图分类号： R378.99;R392-33;R392.13
- 收稿：2018-09-26，
  
  修回：2018-12-09，
  
  纸质出版：2019-02-28
- 稿件说明：
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冯健, 李运明, 刘毓刚, 等. 空肠弯曲菌PEB1的B细胞免疫优势表位的鉴定及保护效果的评价[J]. 第三军医大学学报, 2019,41(4):302-307.

Jian FENG, Yunming LI, Yugang LIU, et al. Fine-mapping of immunodominant linear B-cell epitopes of C.jejuni PEB1 antigen using short overlapping peptides[J]. Journal of Third Military Medical University, 2019, 41(4): 302-307.
冯健, 李运明, 刘毓刚, 等. 空肠弯曲菌PEB1的B细胞免疫优势表位的鉴定及保护效果的评价[J]. 第三军医大学学报, 2019,41(4):302-307. DOI： 10.16016/j.1000-5404.201809180.

Jian FENG, Yunming LI, Yugang LIU, et al. Fine-mapping of immunodominant linear B-cell epitopes of C.jejuni PEB1 antigen using short overlapping peptides[J]. Journal of Third Military Medical University, 2019, 41(4): 302-307. DOI： 10.16016/j.1000-5404.201809180.

摘要

目的

利用表位预测分析技术筛选空肠弯曲菌(

C.jejuni

)黏附蛋白PEB1的B细胞免疫优势表位，并评价其免疫保护效果。

方法

采用氨基酸步移策略，合成18 mer氨基酸重叠肽段。ELISA系统筛选鉴定PEB1的B细胞免疫优势表位。采用KLH偶联免疫优势表位肽免疫BALB/c小鼠，ELISA测定优势表位肽诱导的IgG抗体效价。末次免疫后7 d，经口灌胃感染

C.jejuni

11168，在感染后的28 d，定量检测感染攻毒后各组小鼠的空肠组织

C.jejuni

的定植量以及qRT-PCR技术检测炎性细胞因子TNF-α的相对表达水平。通过HL-60细胞调理吞噬实验测定表位肽诱导产生抗体所介导的调理吞噬杀伤功能。免疫B细胞缺失小鼠，感染攻毒后检测肠组织

C.jejuni

的定植量。

结果

PEB1

55-72aa

、PEB1

97-114aa

、PEB1

211-228aa

均能与PEB1抗血清产生强烈IgG抗体反应。抗PEB1

55-72aa

、抗PEB1

97-114aa

和PEB1

211-228aa

血清均能与重组的PEB1产生较强的抗原抗体反应。与CFA/IFA组相比，免疫PEB1

55-72aa

、PEB1

97-114aa

、PEB1

211-228aa

后血清中的抗体能够显著增强抗体介导的HL-60细胞的调理吞噬作用(

0.01)，均显著降低

C.jejuni

在空肠组织中的定植量，同时也均显著降低炎性细胞因子TNF-α的相对表达水平(

0.01)。PEB1

55-72aa

-KLH+CFA/IFA组，PEB1

97-114aa

-KLH+CFA/IFA组，PEB1

211-228aa

-KLH+CFA/IFA组中，免疫B cell Knock out(B细胞缺失)小鼠攻毒后，

C.jejuni

在空肠组织中的定植量均显著高于WT(野生型)小鼠的定植量(

0.01)。

结论

成功鉴定出3个具有良好免疫原性和免疫保护作用的优势B细胞抗原表位(PEB1

55-72aa

、PEB1

97-114aa

、PEB1

211-228aa

)，可用于

C.jejuni

疫苗的后续开发研究。

Abstract

Objective

To map the immunodominant linear B-cell epitopes of

Campylobacter jejuni

(

C.jejuni

) PEB1 antigen and evaluate the protective immune responses elicited by these epitopes in a mouse model of oral infection with

C.jejuni

Methods

The B-cell immunodominant PEB1 epitopes were identified using synthetic overlapping peptide ELISA. B

ALB/c mice were immunized with the immunodominant PEB1 peptides conjugated with KLH plus CFA/IFA

and the IgG titers against these peptides were detected using ELISA. Seven days after the last immunization

the mice were orally infected with

C.jejuni

11168

and the bacterial burden and expression of tumor necrosis factor-α (TNF-α) mRNA in the jejunum were analyzed using qRT-PCR in 28 d after the challenge. We assessed the effect of the antibodies against the immunodominant PEB1 epitopes in mediating the opsonophagocytic killing of

C.jejuni

by HL-60 cells. We also assessed the bacterial burden in the jejunum following

C.jejuni

challenge in a B cell-knockout mice immunized with the peptides.

Results

The immunodominant peptides PEB1

55-72aa

PEB1

97-114aa

and PEB1

211-228aa

all induced strong IgG responses to PEB1 antiserum

and the antisera of these immunodominant peptides also showed strong IgG responses to recombinant PEB1. Compared with the antiserum of CFA/IFA

the antisera of these immunodominant peptides induced significantly enhanced opsonophagocytic activity of HL-60 cells (

0.01). Both the bacterial burdens and TNF-α mRNA expression level in the jejunum were significantly lowered in the mice immunized with the 3 immunodominant peptides in comparison with the control mice immunized with CFA/IFA (

0.01). In B cell-knockout mice

immunization with the 3 peptides did not provide immune protection against

C.jejuni

and the bacterial burden in the jejunum after

C.jejuni

challenge was significantly greater than that in immunized wild-type mice (

0.01).

Conclusion

We successfully identified 3 linear B-cell epitopes of

C.jejuni

PEB1 (PEB1

55-72aa

PEB1

97-114aa

and PEB1

211-228aa

)

which exhibit good immunogenicity and immuno

protective activities and may facilitate the future development of vaccines against

C.jejuni

infection.

关键词

Keywords

references

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