Immunological and functional characterization of two predict lipoproteins of Leptospira interrogans expressed in the host Escherichia coli

• Grant number: 15/02590-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: September 01, 2015
• End date: January 14, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Ana Lucia Tabet Oller do Nascimento
• Grantee: Priscila Romero Mazzini Pereira
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Leptospirosis is an important zoonosis, recently included in the list of Tropical Neglected Diseases. In urban settings, rodents are the main reservoir of the disease as leptospires colonize their renal tubules and keep shed them through urine. There is no efficient vaccine against the disease. The identification of outer membrane conserved in pathogenic strains is the main goal of the research to elucidate mechanisms involved in pathogenesis. These are the proteins with the potential to interact with external environments and could serve as targets for the host immune system. Among these proteins, lipoproteins are important for new vaccine and diagnostic developments. In this sense, this project proposes the cloning and expression of two predicted lipoproteins identified by bioinfomatics in the genome sequences of Leptospira interrogans. The genes encoding for these lipoproteins, LIC13059 and LIC10879, will be amplified by PCR using complementary primers to the selected sequences. The DNA insert will be cloned into E. coli expression vector. The proteins will be expressed without the signal peptide sequence and in fusion with six histidine resides at the N-terminus. This 6XHis tag will allow the proteins to be purified by nickel charged chromatography. The purified recombinant proteins will be evaluated for their immunogenicity in animal model, reactivity against serum samples of experimentally infected hamsters, checked for their conservation among pathogenic serovars of Leptospira and characterized for their interaction with plasma and host tissues molecules. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SANTOS, JADSON V.; PEREIRA, PRISCILA R. M.; FERNANDES, LUIS G. V.; SIQUEIRA, GABRIELA HASE; DE SOUZA, GISELE O.; SOUZA FILHO, ANTONIO; VASCONCELLOS, SILVIO A.; HEINEMANN, MARCOS B.; CHAPOLA, ERICA G. B.; NASCIMENTO, ANA L. T. O.. Binding of human plasminogen by the lipoprotein LipL46 of Leptospira interrogans. MOLECULAR AND CELLULAR PROBES, v. 37, p. 12-21, FEB 2018. (12/24164-0, 14/50981-0, 15/02590-5)

PEREIRA, PRISCILA R. M.; FERNANDES, LUIS G. V.; DE SOUZA, GISELE O.; VASCONCELLOS, SILVIO A.; HEINEMANN, MARCOS B.; ROMERO, ELIETE C.; NASCIMENTO, ANA L. T. O.. Multifunctional and Redundant Roles of Leptospira interrogans Proteins in Bacterial-Adhesion and fibrin clotting inhibition. INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY, v. 307, n. 6, p. 297-310, SEP 2017. (15/02590-5, 14/50981-0, 12/23913-9, 12/24164-0)

SANTOS, JADSON V.; PEREIRA, PRISCILA R. M.; FERNANDES, LUIS G. V.; SIQUEIRA, GABRIELA HASE; DE SOUZA, GISELE O.; SOUZA FILHO, ANTONIO; VASCONCELLOS, SILVIO A.; HEINEMANN, MARCOS B.; CHAPOLA, ERICA G. B.; NASCIMENTO, ANA L. T. O.. Binding of human plasminogen by the lipoprotein LipL46 of Leptospira interrogans. MOLECULAR AND CELLULAR PROBES, v. 37, p. 10-pg., 2018-02-01. (15/02590-5, 14/50981-0, 12/24164-0)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

PEREIRA, Priscila Romero Mazzini. Immunogenic and functional characterization of two probable lipoproteins of Leptospira interrogans expressed in Escherichia coli.. 2017. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.