Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis
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-...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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2008
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| Online Access: | http://eprints.uanl.mx/15111/1/823.pdf |
| _version_ | 1824414263493525504 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Villatoro Hernández, Julio |
| collection | Repositorio Institucional |
| description | 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. |
| format | Article |
| id | eprints-15111 |
| institution | UANL |
| language | English |
| publishDate | 2008 |
| record_format | eprints |
| spelling | eprints-151112020-06-08T20:42:21Z http://eprints.uanl.mx/15111/ Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis 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 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. 2008 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/15111/1/823.pdf http://eprints.uanl.mx/15111/1.haspreviewThumbnailVersion/823.pdf 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 Rodríguez 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 http://doi.org/10.1186/1475-2859-7-22 doi:10.1186/1475-2859-7-22 |
| spellingShingle | 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 Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| thumbnail | https://rediab.uanl.mx/themes/sandal5/images/online.png |
| title | Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| title_full | Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| title_fullStr | Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| title_full_unstemmed | Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| title_short | Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis |
| title_sort | secretion of biologically active interferon gamma inducible protein 10 ip 10 by lactococcus lactis |
| url | http://eprints.uanl.mx/15111/1/823.pdf |
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