We explored the biology of IL-15 and showed that efficient production of IL-15 is possible only by co-expression in the same cell with the so-called IL-15 Receptor-alpha. We also showed that a second form of IL-15 (SSP IL-15) previously identified in humans and rodents as intracellular or nuclear IL-15 is also efficiently secreted from the cells when co-expressed with the IL-15 Receptor alpha. These results shed new light in the biology and regulation of IL-15 and provide methods for the efficient production and clinical application of this cytokine. Cell lines overproducing soluble bioactive IL-15/IL-15 Receptor alpha heterodimers have been constructed and were used for the production of the bioactive form of IL-15 found in the body. IL-15 purified from over-producing human cells was injected in mice and shown to be bioactive. IL-15 is of interest due to its ability to stimulate the growth, activation and survival of lymphocytes, including CD8 and NK cells. Thus, IL-15 has been considered for cancer immunotherapy and for support of the growth of cytotoxic cell clones after adoptive transfer. Other proposed uses of IL-15 are in lymphopenia, in supporting NK cell growth and activation after NK transfer, and as vaccine adjuvant. We have shown that IL-15 injection accelerates the recovery of lymphocytes in mice rendered lymphopenic after treatment with cytotoxic drugs. We also showed that hetIL-15 can replace the need for lymphodepletion in Adoptive Cell Transfer (ACT), since the injected cells can survive, proliferate and enter the tumors after hetIL-15 treatment. This may have important clinical implications for ACT protocols. We have used the previously developed technologies of RNA optimization to optimize expression of IL-15 cytokine, and have shown that we can over-produce bioactive cytokine after DNA delivery in mice and macaques. DNA delivery of vectors expressing heterodimeric IL-15 leads to systemically active levels of cytokine and the increased proliferation of NK and T cells. We have also used optimized vectors to express IL-12 cytokine in animals. Efficient expression results in bioactive levels, which increase immune response after DNA vaccination, thus becoming important molecular adjuvant for our vaccines. This work established methods to optimize expression of the IL-12 family of cytokines (IL-12, IL-23, IL-27, IL-35). Efficient expression of IL-27 after DNA delivery demonstrated synergy with IL-2 in the elimination of neuroblastoma metastases in mice. We have also developed methods for large-scale isolation and purification of exosomes. We have shown that hetIL-15 is incorporated in exosomes and have produced sufficient quantities for animal experiments. We propose that the versatility of exosomes can be used for the delivery of immunotherapy exosomes to disease sites (tumor or lymphoid tissue). We developed methods for the efficient delivery of exosomes to tumor sites.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010750-12
Application #
9556352
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
12
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Bergamaschi, Cristina; Watson, Dionysios C; Valentin, Antonio et al. (2018) Optimized administration of hetIL-15 expands lymphocytes and minimizes toxicity in rhesus macaques. Cytokine 108:213-224
Watson, Dionysios C; Yung, Bryant C; Bergamaschi, Cristina et al. (2018) Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes. J Extracell Vesicles 7:1442088
Watson, Dionysios C; Moysi, Eirini; Valentin, Antonio et al. (2018) Treatment with native heterodimeric IL-15 increases cytotoxic lymphocytes and reduces SHIV RNA in lymph nodes. PLoS Pathog 14:e1006902
Ng, Sinnie Sin Man; Nagy, Bethany A; Jensen, Shawn M et al. (2017) Heterodimeric IL15 Treatment Enhances Tumor Infiltration, Persistence, and Effector Functions of Adoptively Transferred Tumor-specific T Cells in the Absence of Lymphodepletion. Clin Cancer Res 23:2817-2830
Hong, Enping; Usiskin, Ilana M; Bergamaschi, Cristina et al. (2016) Configuration-dependent Presentation of Multivalent IL-15:IL-15R? Enhances the Antigen-specific T Cell Response and Anti-tumor Immunity. J Biol Chem 291:8931-50
Thaysen-Andersen, M; Chertova, E; Bergamaschi, C et al. (2016) Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. Glycoconj J 33:417-33
Li, Jinyao; Valentin, Antonio; Ng, Sinnie et al. (2015) Differential effects of IL-15 on the generation, maintenance and cytotoxic potential of adaptive cellular responses induced by DNA vaccination. Vaccine 33:1188-96
Bergamaschi, C; Kulkarni, V; Rosati, M et al. (2015) Intramuscular delivery of heterodimeric IL-15 DNA in macaques produces systemic levels of bioactive cytokine inducing proliferation of NK and T cells. Gene Ther 22:76-86
Chertova, Elena; Bergamaschi, Cristina; Chertov, Oleg et al. (2013) Characterization and favorable in vivo properties of heterodimeric soluble IL-15·IL-15R? cytokine compared to IL-15 monomer. J Biol Chem 288:18093-103
Ng, Sinnie Sin Man; Li, Andrew; Pavlakis, George N et al. (2013) Viral infection increases glucocorticoid-induced interleukin-10 production through ERK-mediated phosphorylation of the glucocorticoid receptor in dendritic cells: potential clinical implications. PLoS One 8:e63587

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