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

Villatoro Hernández, Julio y Loera Arias, María de Jesús y Gámez Escobedo, Idalia Analí y Franco Molina, Moisés Armides y Gomez Gutierrez, Jorge G y Rodriguez Rocha, Humberto y Gutiérrez Puente, Yolanda y Saucedo Cárdenas, Odila y Valdés Flores, Jesús y Montes de Oca Luna, Roberto (2008) Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microbial Cell Factories, 7 (1). p. 22. ISSN 1475-2859

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URL o página oficial: http://doi.org/10.1186/1475-2859-7-22

Resumen

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 chemoattract human CD3+ T lymphocytes. This strain of recombinant L. lactis represents a potentially useful tool to be used as a live vaccine in vivo.

Tipo de elemento: Article
Divisiones: Ciencias Biológicas
Medicina
Usuario depositante: Lic. Josimar Pulido
Creadores:
CreadorEmailORCID
Villatoro Hernández, JulioNO ESPECIFICADONO ESPECIFICADO
Loera Arias, María de JesúsNO ESPECIFICADONO ESPECIFICADO
Gámez Escobedo, Idalia AnalíNO ESPECIFICADONO ESPECIFICADO
Franco Molina, Moisés ArmidesNO ESPECIFICADONO ESPECIFICADO
Gomez Gutierrez, Jorge GNO ESPECIFICADONO ESPECIFICADO
Rodriguez Rocha, HumbertoNO ESPECIFICADONO ESPECIFICADO
Gutiérrez Puente, YolandaNO ESPECIFICADONO ESPECIFICADO
Saucedo Cárdenas, OdilaNO ESPECIFICADONO ESPECIFICADO
Valdés Flores, JesúsNO ESPECIFICADONO ESPECIFICADO
Montes de Oca Luna, RobertoNO ESPECIFICADONO ESPECIFICADO
Fecha del depósito: 29 Abr 2019 18:39
Última modificación: 21 Ago 2019 15:03
URI: http://eprints.uanl.mx/id/eprint/15111

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