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Cloning and expression of proteins identified from the genome of the Leptospira interrogans sorovar Copenhageni bacteria with potential for vaccine use and in diagnosis

Grant number: 04/09948-8
Support type:Research Projects - Thematic Grants
Duration: May 01, 2005 - April 30, 2010
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Paulo Lee Ho
Grantee:Paulo Lee Ho
Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Co-Principal Investigators:Ana Lucia Tabet Oller Do Nascimento ; Elizabeth Angelica Leme Martins
Associated scholarship(s):09/06962-3 - Characterization of the SoS response of Leptospira interrogans sorovar Copenhageni, BP.MS
09/02317-6 - Expression and characterization of two surface proteins of Leptospira interrogans, BP.MS
09/02316-0 - Characterization of membrane proteins of Leptospira interrogans expressed in Escherichia coli, BP.MS

Abstract

Leptospirosis is considered the most widespread zoonotic disease caused by the spirochaete from the Leptospira genus. Tn Brazil, 80% of all the diagnosed cases are due to Leptospira interrogans serovar Copenhageni. Rats are the main reservoir in urban centers and the bacteria are disseminated through animal urines. Around 10,000 cases of leptospirosis are annually notified to FUNASA (Health Ministry, Brazil), epidemically associated to the rainy seasons (http://www.funasa.gov.br). A promising way to treat this health problem would be a vaccination program, since the control of rats is a difficult task. Thus, the development of a suitable vaccine against leptospirosis is of seminal importance, since up to date, there is no licensed human vaccine against leptospirosis. Furthermore, the general clinical signs of this disease are often confounded with those of other diseases, like dengue and flu symptoms, reinforcing the need for an efficient diagnose assay for leptospirosis, not yet available in the market. Recently, the genome of the Leptospira interrogans serovar Copenhageni was described. The information of the genome would help to identify potential Leptospira vaccine antigen candidates or antigens for diagnose applications. The present project is divided in 3 subprojects, aiming: 1) cloning of 8 selected hemolysins. Recombinant proteins will be expressed in E. coli and further evaluated for their potential use as vaccine antigens and for potential use in diagnose. Biochemical and immunological characterization will also be performed; 2) cloning of 5 selected lipoproteins. Recombinant proteins will be expressed in E. coli and further evaluated for their potential use as vaccine antigens and for potential use in diagnose. Biochemical and immunological characterization will also be performed; 3) cloning and expression of 5 of the above antigens in Salmonella thyphimurium. The recombinant Salmonella will be evaluated as live vaccine against Leptospira. (AU)