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Villatoro-Hernandez J, Loera-Arias MJ, Gamez-Escobedo A, et al. Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microb Cell Fact. 2008;7:22doi: 10.1186/1475-2859-7-22.
Villatoro-Hernandez, J., Loera-Arias, M. J., Gamez-Escobedo, A., Franco-Molina, M., Gomez-Gutierrez, J. G., Rodriguez-Rocha, H., Gutierrez-Puente, Y., Saucedo-Cardenas, O., Valdes-Flores, J., & Montes-de-Oca-Luna, R. (2008). Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microbial cell factories, 722. https://doi.org/10.1186/1475-2859-7-22
Villatoro-Hernandez, Julio, et al. "Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis." Microbial cell factories vol. 7 (2008): 22. doi: https://doi.org/10.1186/1475-2859-7-22
Villatoro-Hernandez J, Loera-Arias MJ, Gamez-Escobedo A, Franco-Molina M, Gomez-Gutierrez JG, Rodriguez-Rocha H, Gutierrez-Puente Y, Saucedo-Cardenas O, Valdes-Flores J, Montes-de-Oca-Luna R. Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microb Cell Fact. 2008 Jul 28;7:22. doi: 10.1186/1475-2859-7-22. PMID: 18662403; PMCID: PMC2503953.
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Microb Cell Fact. 2008 Jul 28;7:22. doi: 10.1186/1475-2859-7-22.

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

Microbial cell factories

Julio Villatoro-Hernandez, Maria J Loera-Arias, Anali Gamez-Escobedo, Moises Franco-Molina, Jorge G Gomez-Gutierrez, Humberto Rodriguez-Rocha, Yolanda Gutierrez-Puente, Odila Saucedo-Cardenas, Jesus Valdes-Flores, Roberto Montes-de-Oca-Luna

Affiliations

  1. Departamento de Histología, Facultad de Medicina, Universidad Autónoma de Nuevo León (UANL), Monterrey, N,L,, México. [email protected].

PMID: 18662403 PMCID: PMC2503953 DOI: 10.1186/1475-2859-7-22

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 well-characterized 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.

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