Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis

doi:10.1016/S1671-2927(00)8684

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (9): 1517-1527.DOI: 10.1016/S1671-2927(00)8684

Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis

WU Zong-fu, ZHANGWei, WANGYang, LUYan, LUCheng-ping

College of Veterinary Medicine, Nanjing Agricultural University/Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing 210095,P.R.China

收稿日期:2011-05-13 出版日期:2012-09-01 发布日期:2012-09-09
通讯作者: Correspondence LU Cheng-ping, Tel: +86-25-84396517, Fax: +86-25-84396517, E-mail: lucp@njau.edu.cn
作者简介:WU Zong-fu, E-mail: wuzongfu@njau.edu.cn
基金资助:
This work was supported by the fund of the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD), the Natural Science Foundation of Jiangsu Province, China (BK2011644), the Youth Foundation of National Natural Science Foundation of China (31101828), the Youth Foundation of Nanjing Agricultural University, China (KJ2011012), and the Specialized Research Funds for the Doctoral Program of Higher Education of China (20110097120016, 20110097130003).

Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis

WU Zong-fu, ZHANGWei, WANGYang, LUYan, LUCheng-ping

College of Veterinary Medicine, Nanjing Agricultural University/Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing 210095,P.R.China

Received:2011-05-13 Online:2012-09-01 Published:2012-09-09
Contact: Correspondence LU Cheng-ping, Tel: +86-25-84396517, Fax: +86-25-84396517, E-mail: lucp@njau.edu.cn
About author:WU Zong-fu, E-mail: wuzongfu@njau.edu.cn
Supported by:
This work was supported by the fund of the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD), the Natural Science Foundation of Jiangsu Province, China (BK2011644), the Youth Foundation of National Natural Science Foundation of China (31101828), the Youth Foundation of Nanjing Agricultural University, China (KJ2011012), and the Specialized Research Funds for the Doctoral Program of Higher Education of China (20110097120016, 20110097130003).

摘要/Abstract

摘要： Streptococcus suis is a Gram-positive pathogen that causes serious diseases in pigs. In addition to S. suis serotype 2 (SS2), S. suis serotype 9 (SS9) is another prevalent serotype, which is frequently isolated from the organs of diseased pigs in China. An immunoproteomic-based approach was developed to identify antigens common to SS2 and SS9 for vaccine development. Cell wall proteins extracted from SS2 strain HA9801 were screened by two-dimensional Western blot using anti-SS2 sera, anti-SS9 sera, or pre-immune sera pooled from specific pathogen free (SPF) mice. Protein spots on preparative gels were excised and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, which led to the identification of four shared immunogenic proteins (arginine deiminase, translation elongation factor-Ts, o-acetylserine lyase, and 1-phosphofructokinase). The genes encoding these four proteins from SS9 strain GZ0565 were cloned and their proteins were overexpressed in Escherichia coli BL21. Western blot analysis of these recombinant proteins using the convalescent serum of an SPF mini-pig inoculated with the SS2 strain, anti-SS2 sera, and anti-SS9 sera pooled from SPF mice further confirmed the immunogenicity of these proteins. These immunogenic proteins, which are encoded by genes that are reasonably conserved among SS2 and SS9 strains, could be developed as vaccine candidates.

关键词: Streptococcus suis, common antigen, immunoproteomics

Abstract: Streptococcus suis is a Gram-positive pathogen that causes serious diseases in pigs. In addition to S. suis serotype 2 (SS2), S. suis serotype 9 (SS9) is another prevalent serotype, which is frequently isolated from the organs of diseased pigs in China. An immunoproteomic-based approach was developed to identify antigens common to SS2 and SS9 for vaccine development. Cell wall proteins extracted from SS2 strain HA9801 were screened by two-dimensional Western blot using anti-SS2 sera, anti-SS9 sera, or pre-immune sera pooled from specific pathogen free (SPF) mice. Protein spots on preparative gels were excised and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, which led to the identification of four shared immunogenic proteins (arginine deiminase, translation elongation factor-Ts, o-acetylserine lyase, and 1-phosphofructokinase). The genes encoding these four proteins from SS9 strain GZ0565 were cloned and their proteins were overexpressed in Escherichia coli BL21. Western blot analysis of these recombinant proteins using the convalescent serum of an SPF mini-pig inoculated with the SS2 strain, anti-SS2 sera, and anti-SS9 sera pooled from SPF mice further confirmed the immunogenicity of these proteins. These immunogenic proteins, which are encoded by genes that are reasonably conserved among SS2 and SS9 strains, could be developed as vaccine candidates.

Key words: Streptococcus suis, common antigen, immunoproteomics

WU Zong-fu, ZHANGWei, WANGYang, LUYan, LUCheng-ping. Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis[J]. Journal of Integrative Agriculture, 2012, 12(9): 1517-1527.

WU Zong-fu, ZHANGWei , WANGYang , LUYan , LUCheng-ping. Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis[J]. Journal of Integrative Agriculture, 2012, 12(9): 1517-1527.

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Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis

Identification of Antigens Common to Streptococcus suis Serotypes 2 and 9 by ImmunoproteomicAnalysis

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