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

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.

Functional Living Biointerphases

“…Moreover, Lactococcus lactis has been widely used for recombinant therapeutic proteins production, therapeutic drug delivery and vaccine production. [16][17][18][19][20] L. lactis was grown overnight at 30 ºC in M17 medium supplemented with 0.5% glucose in static conditions. After adding the grown culture over the material surface at 30 ° C, we observed that L. lactis adhered, colonized and formed a cocontinuous, stable bacteria multilayer on different material surfaces (polymers, glass, etc.)…”

Enhanced expression of PCV2 capsid protein in Escherichia coli and Lactococcus lactis by codon optimization

“…The use of lactococci as cell factories for the production of bacterial and viral antigens has been achieved by Villatoro-Hernandez et al (2008) and Bahey-El-Din et al (2008). For the development of oral vaccines against PCV2 disease, L. lactis NZ9000, one of the most widely used hosts, was used as a delivery vehicle in the production of Cap protein and its derivatives.…”

Murine interferon-gamma inducible protein-10 (IP-10) secreted by Lactococcus lactis chemo-attracts human CD3+ lymphocytes

“…Protein extraction and sample preparation from L. lactis, SDS-PAGE and Western Blotting were performed as previously reported (Villatoro-Hernandez et al 2008).…”

“…We have previously demonstrated that one strain of L. lactis genetically engineered to express human chemokine lymphotactin (LTN) is able to secrete at least 500 ng per ml of culture in 1 h (Zavala-Flores et al 2009). We also demonstrated that human interferon-gamma inducible protein-10 (IP-10) from a different strain of L. lactis genetically manipulated to express and secrete this human chemokine is able to chemo-attract human T lymphocytes (Villatoro-Hernandez et al 2008). This chemokine has been the focus of many studies due to its antitumoral activity in different types of tumor cells (Frederick and Clayman 2001).…”