{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T06:00:30Z","timestamp":1762063230311,"version":"build-2065373602"},"reference-count":42,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2022,8,5]],"date-time":"2022-08-05T00:00:00Z","timestamp":1659657600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MCIN\/AEI\/10.13039\/501100011033\/","award":["B31_20R"],"award-info":[{"award-number":["B31_20R"]}]},{"DOI":"10.13039\/501100010067","name":"overnment of Aragon","doi-asserted-by":"publisher","award":["B31_20R"],"award-info":[{"award-number":["B31_20R"]}],"id":[{"id":"10.13039\/501100010067","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Nine kDa granulysin (GRNLY) is a human cytolytic protein secreted by cytotoxic T lymphocytes (CTL) and NK cells of the immune system whose demonstrated physiological function is the elimination of bacteria and parasites. In previous studies by our group, the anti-tumor capacity of recombinant granulysin was demonstrated, both in vitro and in vivo. In the present work, we developed lipid nanoparticles whose surfaces can bind recombinant granulysin through the formation of a complex of coordination between the histidine tail of the protein and Ni2+ provided by a chelating lipid in the liposome composition and termed them LUV-GRNLY, for granulysin-bound large unilamellar vesicles. The objective of this formulation is to increase the granulysin concentration at the site of contact with the target cell and to increase the cytotoxicity of the administered dose. The results obtained in this work indicate that recombinant granulysin binds to the surface of the liposome with high efficiency and that its cytotoxicity is significantly increased when it is in association with liposomes. In addition, it has been demonstrated that the main mechanism of death induced by both granulysin and LUV-GRNLY is apoptosis. Jurkat-shBak cells are resistant to GRNLY and also to LUV-GRNLY, showing that LUV-GRNLY uses the mitochondrial apoptotic pathway to induce cell death. On the other hand, we show that LUV-GRNLY induces the expression of the pro-apoptotic members of the Bcl-2 family Bim and especially PUMA, although it also induced the expression of anti-apoptotic Bcl-xL. In conclusion, we demonstrate that binding of GRNLY to the surfaces of liposomes clearly augments its cytotoxic potential, with cell death executed mainly by the mitochondrial apoptotic pathway.<\/jats:p>","DOI":"10.3390\/ijms23158705","type":"journal-article","created":{"date-parts":[[2022,8,7]],"date-time":"2022-08-07T22:51:46Z","timestamp":1659912706000},"page":"8705","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Conjugation of the 9-kDa Isoform of Granulysin with Liposomes Potentiates Its Cytotoxicity"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3153-389X","authenticated-orcid":false,"given":"Ruth","family":"Soler-Agesta","sequence":"first","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1657-4792","authenticated-orcid":false,"given":"Patricia","family":"Guerrero-Ochoa","sequence":"additional","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1710-4821","authenticated-orcid":false,"given":"Joaqu\u00edn","family":"Marco-Brualla","sequence":"additional","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raquel","family":"Ib\u00e1\u00f1ez-P\u00e9rez","sequence":"additional","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2315-9079","authenticated-orcid":false,"given":"Isabel","family":"Marzo","sequence":"additional","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis","family":"Mart\u00ednez-Lostao","sequence":"additional","affiliation":[{"name":"Nanoscience Institute of Aragon (INA), 50018 Zaragoza, Spain"},{"name":"Immunology Department, Clinical University Hospital \u201cLozano Blesa\u201d, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5175-8394","authenticated-orcid":false,"given":"Alberto","family":"Anel","sequence":"additional","affiliation":[{"name":"Apoptosis, Immunity & Cancer Group, Department of Biochemistry and Molecular and Cell Biology, Faculty of Sciences, Arag\u00f3n Health Research Institute (IIS Arag\u00f3n), University of Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6119","DOI":"10.4049\/jimmunol.1200570","article-title":"15 kDa Granulysin Causes Differentiation of Monocytes to Dendritic Cells but Lacks Cytotoxic Activity","volume":"188","author":"Clayberger","year":"2012","journal-title":"J. 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