Long-term maintenance of peripheral blood derived human NK cells in a novel human IL-15- transgenic NOG mouse

Abstract

We generated a novel mouse strain expressing transgenic human interleukin-15 (IL-15) using the severe immunodeficient NOD/Shi-scid-IL-2Rγnull (NOG) mouse genetic background (NOG-IL-15 Tg). Human natural killer (NK) cells, purified from the peripheral blood (hu-PB-NK) of normal healthy donors, proliferated when transferred into NOG-IL-15 Tg mice. In addition, the cell number increased, and the hu-PB-NK cells persisted for 3 months without signs of xenogeneic graft versus host diseases (xGVHD). These in vivo-expanded hu-PB-NK cells maintained the original expression patterns of various surface antigens, including NK receptors and killer cell immunoglobulin-like receptor (KIR) molecules. They also contained significant amounts of granzyme A and perforin. Inoculation of K562 leukemia cells into hu-PB-NK-transplanted NOG-IL-15 Tg mice resulted in significant suppression of tumor growth compared with non-transplanted mice. Furthermore, NOG-IL-15 Tg mice allowed for engraftment of in vitro-expanded NK cells prepared for clinical cell therapy. These cells exerted antibody-dependent cell-mediated cytotoxicity (ADCC) on Her2-positive gastric cancer cells in the presence of therapeutic anti-Her2 antibody, and subsequently suppressed tumor growth. Our results collectively suggest that the NOG-IL-15 Tg mice are a useful model for studying human NK biology and evaluating human NK cell-mediated in vivo cytotoxicity.

NK cells prevent the formation of teratomas derived from human induced pluripotent stem cells

“…Our results also demonstrated that inhibition of teratomas formation was possible only if hiPSCs were injected prior to the adoptive transfer of total PBMCs or purified NK cells, consistent with the prior observation that hiPSCs-derived myoblasts are not the target of NK cells in vitro and in vivo (Benabdallah et al submitted). Moreover, in the absence of recombinant human IL-15 to support the proliferation of NK in our mice [14], our results also suggest that the relatively low number of NK cells injected during the adoptive transfer procedure was sufficient to prevent the growth of teratomas. These results suggest that if immunosuppressive drugs were to be used to increase engraftment of iPSC-derived cells, it would be very important to insure no inhibition of the NK cell compartment to lower the risk of forming a teratoma.…”

Humanization of Immunodeficient Animals for the Modeling of Transplantation, Graft Versus Host Disease, and Regenerative Medicine

“…The severity of immunodeficiency has been increased by combining mutations in the aforementioned genes and/or Prkdc. 167 , 174 , 176 , 177 …”

“… 128 Additionally, Prkdc mutations to obtain a SCID phenotype in many mouse strains (like all NOD-derived immunodeficient strains such as NSG and NOG) increase toxicity due to irradiation in certain models, such as in cancer treatments, since PRKDC is an enzyme essential in DNA repair and its absence generates uncontrolled toxicity in the host tissues. Some 174 , 176 , 182 but not all 177 immunodeficient rats also carry a mutation in the Prkdc gene.…”

“…Animals from the RRG line, with Rag1 and Il2rg mutations, indefinitely accepted human skin, tumors, and even hepatocytes, but did not accept human PBMCs. 177 However, RRG animals crossed with rats expressing human SIRPα 207 (RRGS animals) or with macrophage depletion allowed immune humanization using human PBMC. 128 These rats humanized with PBMCs could develop GvHD and reject tumor cells.…”

The Utility of Human Immune System Mice for High-Containment Viral Hemorrhagic Fever Research

“…Mice transgenic for human cytokine genes have improved development of cell lineages reliant on cytokines that are not cross-reactive between mice and humans. For example, mice transgenic for human GM-CSF, IL-3, and SCF support greater development of human myeloid lineages [16], whereas mice transgenic for human IL-2 [17] and IL-15 [18] support better NK cell development and survival. Functionality, as opposed to levels of reconstitution, has also been improved through transgenic expression of human leukocyte antigens (HLA) in graft recipient mouse strains.…”