OBJECTIVE: Anti-tumor effects mediated by adoptively transferred natural killer (NK) cells are dependent on the presence of interleukin-2 (IL-2). IL-2 is considered to be a survival factor for NK cells and an enhancer of their cytotoxic potential. However, systemic administration of IL-2 is frequently impeded by undesirable side effects, such as high toxicity and nonlocalized administration. Genetic modification of NK cells expressing IL-2 in a localized and controlled manner could be a powerful tool for overcoming these obstacles. METHODS: Consequently, we have cloned the IL-2 gene using PCR and designed constructs that target IL-2 to specific subcellular compartments. The IL-2-dependent NK-92 cell line was used to verify the functionality of the subcellularly targeted IL-2 constructs. RESULTS: IL-2 targeted specifically to the endoplasmic reticulum (ER) was sufficient to support growth of NK-92 cells. In such cell lines, IL-2 was verified to be localized to the ER. IL-2 was not detected in the supernatant and growth of non-IL-2-modified NK-92 cells was not supported during coculturing experiments. IL-2-transduced NK-92 cell lines showed comparable functional activity and cytotoxicity to parental NK-92 cells. CONCLUSION: We demonstrate the ability of ER-retained IL-2 to provide autocrine growth stimulation to NK-92 cells, without secretion of the cytokine to the extracellular compartment. Therapy with IL-2 gene-modified autoactivating NK cells may avoid side effects imposed by exogenously administered IL-2.
OBJECTIVE: Anti-tumor effects mediated by adoptively transferred natural killer (NK) cells are dependent on the presence of interleukin-2 (IL-2). IL-2 is considered to be a survival factor for NK cells and an enhancer of their cytotoxic potential. However, systemic administration of IL-2 is frequently impeded by undesirable side effects, such as high toxicity and nonlocalized administration. Genetic modification of NK cells expressing IL-2 in a localized and controlled manner could be a powerful tool for overcoming these obstacles. METHODS: Consequently, we have cloned the IL-2 gene using PCR and designed constructs that target IL-2 to specific subcellular compartments. The IL-2-dependent NK-92 cell line was used to verify the functionality of the subcellularly targeted IL-2 constructs. RESULTS:IL-2 targeted specifically to the endoplasmic reticulum (ER) was sufficient to support growth of NK-92 cells. In such cell lines, IL-2 was verified to be localized to the ER. IL-2 was not detected in the supernatant and growth of non-IL-2-modified NK-92 cells was not supported during coculturing experiments. IL-2-transduced NK-92 cell lines showed comparable functional activity and cytotoxicity to parental NK-92 cells. CONCLUSION: We demonstrate the ability of ER-retained IL-2 to provide autocrine growth stimulation to NK-92 cells, without secretion of the cytokine to the extracellular compartment. Therapy with IL-2 gene-modified autoactivating NK cells may avoid side effects imposed by exogenously administered IL-2.
Authors: Jose M Ayuso; Regan Truttschel; Max M Gong; Mouhita Humayun; Maria Virumbrales-Munoz; Ross Vitek; Mildred Felder; Stephen D Gillies; Paul Sondel; Kari B Wisinski; Manish Patankar; David J Beebe; Melissa C Skala Journal: Oncoimmunology Date: 2018-12-12 Impact factor: 8.110
Authors: Carly E Whyte; Kailash Singh; James Dooley; Oliver T Burton; Meryem Aloulou; Lubna Kouser; Rafael Valente Veiga; Amy Dashwood; Hanneke Okkenhaug; Samira Benadda; Alena Moudra; Orian Bricard; Stephanie Lienart; Pascal Bielefeld; Carlos P Roca; Francisco José Naranjo-Galindo; Félix Lombard-Vadnais; Steffie Junius; David Bending; Masahiro Ono; Tino Hochepied; Timotheus Y F Halim; Susan Schlenner; Sylvie Lesage; Adrian Liston Journal: J Exp Med Date: 2022-06-14 Impact factor: 17.579
Authors: Chantiya Chanswangphuwana; David S J Allan; Mala Chakraborty; Robert N Reger; Richard W Childs Journal: Mol Ther Date: 2020-09-20 Impact factor: 11.454