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Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis

Authors :
Villatoro Hernández, Julio
Loera Arias, María de Jesús
Gámez Escobedo, Idalia Analí
Franco Molina, Moisés Armides
Gomez Gutierrez, Jorge G
Rodríguez Rocha, Humberto
Gutiérrez Puente, Yolanda
Saucedo Cárdenas, Odila
Valdés Flores, Jesús
Montes de Oca Luna, Roberto
Villatoro Hernández, Julio
Loera Arias, María de Jesús
Gámez Escobedo, Idalia Analí
Franco Molina, Moisés Armides
Gomez Gutierrez, Jorge G
Rodríguez Rocha, Humberto
Gutiérrez Puente, Yolanda
Saucedo Cárdenas, Odila
Valdés Flores, Jesús
Montes de Oca Luna, Roberto
Publication Year :
2008

Abstract

Background: Chemokines are a large group of chemotactic cytokines that regulate and direct migration of leukocytes, activate inflammatory responses, and are involved in many other functions including regulation of tumor development. Interferon-gamma inducible-protein-10 (IP-10) is a member of the C-X-C subfamily of the chemokine family of cytokines. IP-10 specifically chemoattracts activated T lymphocytes, monocytes, and NK cells. IP-10 has been described also as a modulator of other antitumor cytokines. These properties make IP-10 a novel therapeutic molecule for the treatment of chronic and infectious diseases. Currently there are no suitable live biological systems to produce and secrete IP-10. Lactococcus lactis has been wellcharacterized over the years as a safe microorganism to produce heterologous proteins and to be used as a safe, live vaccine to deliver antigens and cytokines of interest. Here we report a recombinant strain of L. lactis genetically modified to produce and secrete biologically active IP-10. Results: The IP-10 coding region was isolated from human cDNA and cloned into an L. lactis expression plasmid under the regulation of the pNis promoter. By fusion to the usp45 secretion signal, IP-10 was addressed out of the cell. Western blot analysis demonstrated that recombinant strains of L. lactis secrete IP-10 into the culture medium. Neither degradation nor incomplete forms of IP-10 were detected in the cell or supernatant fractions of L. lactis. In addition, we demonstrated that the NICE (nisin-controlled gene expression) system was able to express IP-10 "de novo" even two hours after nisin removal. This human IP-10 protein secreted by L. lactis was biological active as demonstrated by Chemotaxis assay over human CD3+T lymphocytes. Conclusion: Expression and secretion of mature IP-10 was efficiently achieved by L. lactis forming an effective system to produce IP-10. This recombinant IP-10 is biologically active as demonstrated by its ability to ch

Details

Database :
OAIster
Notes :
text, English
Publication Type :
Electronic Resource
Accession number :
edsoai.on1105320259
Document Type :
Electronic Resource