title: Liver gene transfer of interkeukin-15 constructs that become part of circulating high density lipoproteins for immunotherapy.
abstract: Apolipoprotein A-I (Apo A-I) is a major component of high density lipoproteins (HDL) that transport cholesterol in circulation. We have constructed an expression plasmid encoding a chimeric molecule encompassing interleukin-15 (IL-15) and Apo A-I (pApo-hIL15) that was tested by hydrodynamic injections into mice and was co-administered with a plasmid encoding the sushi domain of IL-15Rα (pSushi) in order to enhance IL-15 trans-presentation and thereby bioactivity. The pharmacokinetics of the Apo A-I chimeric protein were much longer than non-stabilized IL-15 and its bioactivity was enhanced in combination with IL-15Rα Sushi. Importantly, the APO-IL-15 fusion protein was incorporated in part into circulating HDL. Liver gene transfer of these constructs increased NK and memory-phenotype CD8 lymphocyte numbers in peripheral blood, spleen and liver as a result of proliferation documented by CFSE dilution and BrdU incorporation. Moreover, the gene transfer procedure partly rescued the NK and memory T-cell deficiency observed in IL-15Rα(-/-) mice. pApo-hIL15+ pSushi gene transfer to the liver showed a modest therapeutic activity against subcutaneously transplanted MC38 colon carcinoma tumors, that was more evident when tumors were set up as liver metastases. The improved pharmacokinetic profile and the strong biological activity of APO-IL-15 fusion protein holds promise for further development in combination with other immunotherapies.
authors.affiliation: Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Ochoa
authors.forename: Jessica
authors.initials: J
authors.lastname: Fioravanti
authors.forename: Erwin H
authors.initials: EH
authors.lastname: Duitman
authors.forename: Jose
authors.initials: J
authors.lastname: Medina-Echeverz
authors.forename: Asis
authors.initials: A
authors.lastname: Palazon
authors.forename: Ainhoa
authors.initials: A
authors.lastname: Arina
authors.forename: Juan
authors.initials: J
authors.lastname: Dubrot
authors.forename: Carlos
authors.initials: C
authors.lastname: Alfaro
authors.forename: Aizea
authors.initials: A
authors.lastname: Morales-Kastresana
authors.forename: Oihana
authors.initials: O
authors.lastname: Murillo
authors.forename: Sandra
authors.initials: S
authors.lastname: Hervas-Stubbs
authors.forename: Jesus
authors.initials: J
authors.lastname: Prieto
authors.forename: Pedro
authors.initials: P
authors.lastname: Berraondo
authors.forename: Ignacio
authors.initials: I
authors.lastname: Melero
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000097:blood; D015153:Blotting, Western; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002452:Cell Count; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D007156:Immunologic Memory; D007167:Immunotherapy; D007279:Injections, Subcutaneous; D019409:Interleukin-15 / Q000097:blood / Q000235:genetics* / Q000627:therapeutic use*; D053720:Interleukin-15 Receptor alpha Subunit / Q000737:chemistry / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology; D008075:Lipoproteins, HDL / Q000097:blood*; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009362:Neoplasm Metastasis; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000627:therapeutic use; D013154:Spleen / Q000378:metabolism
chemical_list: D016632:Apolipoprotein A-I; D019409:Interleukin-15; D053720:Interleukin-15 Receptor alpha Subunit; D008075:Lipoproteins, HDL; D011994:Recombinant Proteins