title: Membrane-attached Cytokines Expressed by mRNA Electroporation Act as Potent T-Cell Adjuvants.
abstract: Proinflammatory cytokines are widely explored in different adoptive cell therapy protocols for enhancing survival and function of the transferred T cells, but their systemic administration is often associated with severe toxicity which limits their clinical use. To confine cytokine availability to the therapeutic T cells, we expressed 3 key cytokines, IL-2, IL-12, and IL-15, as integral T-cell membrane proteins. To prevent permanent activation of growth signaling pathways, we delivered these genes to T cells through mRNA electroporation. The engineered cytokines could be detected on the surface of mRNA-transfected cells and binding to their cell-surface receptors mainly occurred in cis. The 3 human cytokines supported the ex vivo growth of activated human CD8 and CD4 T cells for at least 6 days posttransfection, comparably to high-dose soluble IL-2. Similarly, membrane IL-2, membrane IL-12, and, to a lesser extent, membrane IL-15, were comparable with their soluble counterparts in supporting proliferation of splenic mouse CD8 T cells. Following electroporation of human CD8 T cells and antimelanoma tumor-infiltrating lymphocytes, membrane cytokines synergized with constitutively active toll-like receptor 4 in inducing interferon-γ secretion. Efficient cooperation with TLR4 was also evident in the upregulation of the activation molecules CD25, CD69, CD137 (4-1BB), and CD134 (OX40). Taken together, membrane cytokines expressed through mRNA transfection emerge as effective tools for enhancing T-cell proliferation and function and may have potential use in adoptive T-cell therapy.
authors: *Laboratory of Immunology, MIGAL-Galilee Research Institute, Kiryat Shmona †Sharett Institute of Oncology, Hadassah Hebrew University Hospital, Jerusalem ‡Ella Lemelbaum Institute for Melanoma, Sheba Medical Center, Ramat Gan §Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Tel Aviv ∥Tel-Hai College, Upper Galilee, Israel.
authors: Hadas
authors: H
authors: Weinstein-Marom
authors: Aviad
authors: A
authors: Pato
authors: Noam
authors: N
authors: Levin
authors: Keren
authors: K
authors: Susid
authors: Orit
authors: O
authors: Itzhaki
authors: Michal J
authors: MJ
authors: Besser
authors: Tamar
authors: T
authors: Peretz
authors: Alon
authors: A
authors: Margalit
authors: Michal
authors: M
authors: Lotem
authors: Gideon
authors: G
authors: Gross
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D018274:Electroporation; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D015202:Protein Engineering; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology* / Q000637:transplantation; D051197:Toll-Like Receptor 4 / Q000378:metabolism*
chemical_list: D016207:Cytokines; D012333:RNA, Messenger; D051197:Toll-Like Receptor 4