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Identification of immunoreactive proteins of Brucella melitensis by immunoproteomics

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Science China(Life Sciences) 2011年第9期54卷 880-887页

作者： ZHAO ZhongPeng YAN Fang JI WenHui LUO DeYan LIU Xin XING Li DUAN YueQiang YANG PengHui SHI XiuMin LI Zhong WANG XiLiang State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology Beijing 100071 China College of Animal Science and Veterinary Medicine Shanxi Agricultural University Taigu 030801 China

Infection with Brucella causes brucellosis, a chronic disease in humans, which induces abortion and sterility in livestock. Among the different Brucella species, Brucella melitensis is considered the most virulent and is the predominant species associated with outbreaks in China. To date, no safe human vaccine is available against Brucella infection. The currently used live vaccines against Brucella in livestock induce antibodies that interfere with the diagnosis of field infection in vaccinated ani- mals, which is harmful to eradication programs. However, there is as yet no complete profile of immunogenic proteins of B. melitensis. Towards the development of a safer, equally efficacious, and field infection-distinguishable vaccine, we used immunoproteomics to identify novel candidate immunogenic proteins from B. melitensis M5. Eighty-eight immunoreactive protein spots from B. melitensis M5 were identified by Western blotting and were assigned to sixty-one proteins by mass spectrometry, including many new immunoreactive proteins such as elongation factor G, FOFI ATP synthase subunit beta, and OMPI. These provide many candidate immunoreactive proteins for vaccine development.

关键词： Brucella immunoproteomics immunoreactive protein

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

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Journal of Integrative Agriculture 2012年第9期11卷 1517-1527页

作者： WU Zong-fu ZHANG Wei WANG Yang LU Yan LU Cheng-ping College of Veterinary Medicine Nanjing Agricultural University/Key Lab of Animal Bacteriology Ministry of Agriculture Nanjing 210095P.R.China

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

Immunoproteomic Assay of Antigenic Surface Proteins in Streptococcus equi ssp. zooepidemicus

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Agricultural Sciences in China 2011年第7期10卷 1096-1105页

作者： MAO Ying FAN Hong-jie ZHOU Yong-hua LU Cheng-ping Key Laboratory of Animal Disease Diagnostic and Immunology Ministry of Agriculture/College of Veterinary Medicine Nanjing Agricultural University Nanjing 210095 P.R.China Jiangsu Institute of Parasitic Diseases Wuxi 214064 P.R.China

Streptococcus equi ssp. zooepidemicus （S. zooepidemicus） is a zoonotic pathogen with worldwide distribution. Lacking suitable vaccine and virulent maker is still bottleneck to control this infection. An immunoproteomic approach has been used to screen the membrane-associated and cell wall-associated proteins of S. zooepidemicus isolate in China CY to discover vaccine candidate antigens and therapeutic agents. Finally, 11 membrane-associated proteins, and 13 cell wall-associated proteins were successfully identified. BLAST （***） results also indicated that nucleotide sequences of majority identified proteins shared high homology （60%） with S. zooepidemicus, except for AC1-3, AC5, AC8, and AC13. Moreover, genes for 7 of the identified proteins were detected from CY; compared with ST171, 3 of them （AM1, AM8 and AC11） were only found in virulent strains （CY）. All of the proteins identified in this study remain not to be reported in S. zooepidemicus. Some of the proteins serve a vital role in the immune system and reproduction of host species according to available data, while the functions of the rest were seldom researched.

关键词： membrane-associated proteins (MAP) cell wall-associated proteins (CP) immunoproteomics S. zooepidemicus

Study roadmap for high-throughput development of easy to use and affordable biomarkers as diagnostics for tropical diseases:a focus on malaria and schistosomiasis

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Infectious Diseases of Poverty 2017年第1期6卷 1149-1156页

作者： Kokouvi Kassegne Ting Zhang Shen-Bo Chen Bin Xu Zhi-Sheng Dang Wang-Ping Deng Eniola Michael Abe Hai-Mo Shen Wei Hu Takele Geressu Guyo Solomon Nwaka Jun-Hu Chen Xiao-Nong Zhou National Institute of Parasitic Diseases(NIPD) Chinese Centre for Disease Control and PreventionShanghai 200025People’s Republic of China WHO Collaborating Centre for Tropical Diseases National Centre for International Research on Tropical DiseasesKey Laboratory of Parasite and Vector Biology of the Chinese Ministry of HealthShanghai 200025People’s Republic of China Department of Microbiology and Microbial Engineering School of Life ScienceFudan UniversityShanghai 200433People’s Republic of China African Network for Drugs&Diagnostics Innovation(ANDI) Addis AbabaEthiopia

Background:Interventions are currently being used against‘infectious diseases of poverty’,which remain highly debilitating and deadly in most endemic countries,especially malaria,schistosomiasis,echinococcosis and African sleeping ***,major limitations of current‘traditional’methods for diagnosis are neither simple nor convenient for population surveillance,and showed low sensitivity and *** to novel technologies for the development of adequate and reliable tools are expressly needed.A collaborative project between African Network for Drugs and Diagnostics Innovation and partner institutions in Africa and China aims to screen suitable serological biomarkers for diagnostic pipelines against these‘diseases of the poor’.Methods:Parasite-specific exposed versus unexposed individuals were screened and sera or urine/stools were collected through case-control studies in China and African *** genes/open reading frames were selected,then will be cloned and cell-free expressed,quantified and *** antigens/epitopes will be probed and screened with sera from exposed or unexposed individuals using a high-throughput antigen screening platform as the study *** specificity and sensitivity of highly immunoreactive biomarkers will be evaluated as well,using enzyme-linked immunosorbent assays or ***:This roadmap explicitly unfolds the integrated operating procedures with focus on malaria and schistosomiasis,for the identification of suitable biomarkers that will aid the prioritization of diagnostics for population ***,there is need to further validate any new diagnostic through comparison with standard methods in field deployable tests for each *** expectations for the future are to seek regulatory approval and promote the use of diagnostics in endemic areas.

关键词： Malaria Schistosomiasis Neglected tropical diseases immunoproteomics Protein microarray Serological biomarker Diagnosis

Overexpression of Annexin A2 Is Associated with Abnormal Ubiquitination in Breast Cancer

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Genomics, Proteomics & Bioinformatics 2012年第3期10卷 153-157页

作者： Shishan Deng Baoqian Jing Tianyong Xing Lingmi Hou Zhengwei Yang Department of Anatomy North Sichuan Medical CollegeNanchong 637007China Institute of Immunology and Molecular Biology North Sichuan Medical CollegeNanehong 637007China Department of General Surgery The Affiliated Hospital of North Sichuan Medical CollegeNanchong 637007China Morphometric Research Laboratory North Sichuan Medical CollegeNanchong 637007China

Abnormal expression of annexin A2 contributes to metastasis and infiltration of cancer cells. To elucidate the cause of abnormal expres- sion of annexin A2, Western blotting, immunoproteomics and immunohistochemical staining were performed to analyze differentially ubiquitinated proteins between fresh breast cancer tissue and its adjacent normal breast tissue from five female volunteers. We detected an ubiquitinated protein that was up-regulated in the cancer tissue, which was further identified as annexin A2 by mass spectrometry. These results suggest that abnormal ubiquitination and/or degradation of annexin A2 may lead to presence of annexin A2 at high level, which may further promote metastasis and infiltration of the breast cancer cells.

关键词： Annexin A2 Breast cancer immunoproteomics Ubiquitination