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However, both cell-extrinsic and intrinsic mechanisms often diminish the in-vivo potency of these therapeutic T cells, limiting their clinical efficacy and broader use. Direct activation of human T cells by Toll-like receptor (TLR) ligands induces T cell survival and proliferation, boosts the production of proinflammatory cytokines and augments resistance to regulatory T cell (Treg) suppression. Removal of the TLR ligand-binding region results in constitutive signalling triggered by the remaining cytosolic Toll\/interleukin-1 receptor (TIR) domain. The use of such TIR domains therefore offers an ideal means for equipping anti-tumour T cells with the arsenal of functional attributes required for improving current clinical protocols. Here we show that constitutively active (ca)TLR-4 can be expressed efficiently in human T cells using mRNA electroporation. The mere expression of caTLR-4 mRNA in polyclonal CD8 and CD4 T cells induced the production of interferon (IFN)-\u03b3, triggered the surface expression of CD25, CD69 and 4-1BB and up-regulated a panel of cytokines and chemokines. In tumour-infiltrating lymphocytes prepared from melanoma patients, caTLR-4 induced robust IFN-\u03b3 secretion in all samples tested. Furthermore, caTLR-4 enhanced the anti-melanoma cytolytic activity of tumour-infiltrating lymphocytes and augmented the secretion of IFN-\u03b3, tumour necrosis factor (TNF)-\u03b1 and granulocyte\u2013macrophage colony-stimulating factor (GM-CSF) for at least 4 days post-transfection. Our results demonstrate that caTLR-4 is capable of exerting multiple T cell-enhancing effects and can potentially be used as a genetic adjuvant in adoptive cell therapy.<\/jats:p>","DOI":"10.1111\/cei.12688","type":"journal-article","created":{"date-parts":[[2015,7,23]],"date-time":"2015-07-23T23:37:22Z","timestamp":1437694642000},"page":"220-229","source":"Crossref","is-referenced-by-count":16,"title":["Messenger RNA encoding constitutively active Toll-like receptor 4 enhances effector functions of human T cells"],"prefix":"10.1093","volume":"182","author":[{"given":"A","family":"Pato","sequence":"first","affiliation":[{"name":"Laboratory of Immunology, MIGAL \u2013 Galilee Research Institute, Kiryat, Shmona"},{"name":"Sharett Institute of Oncology, Hadassah Hebrew University Hospital, Jerusalem"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G","family":"Eisenberg","sequence":"additional","affiliation":[{"name":"Sharett Institute of Oncology, Hadassah Hebrew University Hospital, Jerusalem"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Machlenkin","sequence":"additional","affiliation":[{"name":"Sharett Institute of Oncology, Hadassah Hebrew University Hospital, Jerusalem"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Margalit","sequence":"additional","affiliation":[{"name":"Laboratory of Immunology, MIGAL \u2013 Galilee Research Institute, Kiryat, Shmona"},{"name":"Department of Biotechnology, Tel-Hai College, Upper, Galilee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G","family":"Cafri","sequence":"additional","affiliation":[{"name":"Laboratory of Immunology, MIGAL \u2013 Galilee Research Institute, Kiryat, Shmona"},{"name":"Department of Immunology, Weizmann Institute of Science, Rehovot, Israel"},{"name":"Surgery Branch, National Cancer Institute, Bethesda, Maryland, 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