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Animals were sacrificed 3 and 7 days post-RFA. Outcomes included immunohistochemistry of dendritic cells (DCs), M1 and M2 macrophages, T-helper cells (Th1) (CD4<jats:sup>+<\/jats:sup>), cytotoxic T- lymphocytes (CTL) (CD8<jats:sup>+<\/jats:sup>), T-regulator cells (T-reg) (FoxP3<jats:sup>+<\/jats:sup>) and Fas Ligand activated CTLs (Fas-L<jats:sup>+<\/jats:sup>) in the periablational rim and untreated index tumor. M1\/M2, CD4<jats:sup>+<\/jats:sup>\/CD8<jats:sup>+<\/jats:sup> and CD8<jats:sup>+<\/jats:sup>\/FoxP3<jats:sup>+<\/jats:sup> ratios were calculated. Phase II, 40 rats with double tumors were randomized to 4 groups: a) sham, b) RFA, c) RFA-BL and d) RFA-lip-GM-CSF. Synchronous untreated tumors collected at 7d were analyzed similarly.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Results<\/jats:title>\n<jats:p>RFA-lip-GMCSF increased periablational M1, CTL and CD8<jats:sup>+<\/jats:sup>\/FoxP3<jats:sup>+<\/jats:sup> ratio at 3 and 7d, and activated CTLs 7d post-RFA (p&lt;0.05). RFA-lip-GMSCF also increased M2, T-reg, and reduced CD4<jats:sup>+<\/jats:sup>\/CD8<jats:sup>+<\/jats:sup> 3 and 7d post-RFA respectively (p&lt;0.05). In untreated index tumor, RFA-lip-GMCSF improved DCs, M1, CTLs and activated CTL 7d post-RFA (p&lt;0.05). Furthermore, RFA-lip-GMSCF increased M2 at 3 and 7d, and T-reg 7d post-RFA (p&lt;0.05). In synchronous tumors, RFA-BL and RFA-lip-GM-CSF improved DC, Th1 and CTL infiltration 7d post-RFA.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>Systemic liposomal GM-CSF combined with RFA improves intratumoral immune cell trafficking, specifically populations initiating (DC, M1) and executing (CTL, FasL<jats:sup>+<\/jats:sup>) anti-tumor immunity. Moreover, liposomes influence synchronous untreated metastases increasing Th1, CTL and DCs infiltration.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0293141","type":"journal-article","created":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T17:25:14Z","timestamp":1698341114000},"page":"e0293141","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Combined thermal ablation and liposomal granulocyte-macrophage colony stimulation factor increases immune cell trafficking in a small animal tumor model"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3462-7007","authenticated-orcid":true,"given":"Marwan","family":"Moussa","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Md. 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