Generation of induced pluripotent stem cell-derived mice by reprogramming of a mature NKT cell

Abstract

The successful generation of iPSC-derived mouse strains to study NKT cells

Influences of age and sex on leukocytes of healthy horses and their ex vivo cytokine release

“…Medium settings were used to evaluate spontaneous cytokine release and PMA/Ionomycin stimulation to assess cytokine induction. This concentration of stimuli was used in recent studies on human PBMC (Nagendran et al, 2013), T cells (Rohlman et al, 2012), murine T cells (Edwards et al, 2014), murine lung cells (Mays et al, 2013) and natural killer T cells (Ren et al, 2014) and had proven useful to induce all targeted cytokines (except IFN␣) in preliminary experiments with equine PBMC (data not shown). After incubation, cell-free supernatants were obtained and stored at −80 • C until further analysis.…”

New Genetically Manipulated Mice Provide Insights Into the Development and Physiological Functions of Invariant Natural Killer T Cells

“…These studies demonstrate the feasibility of expanding functionally competent iNKT cells via an iPSC phase, an approach that may be adapted for iNKT cell-targeted therapy in humans ( 56 , 57 ). We further succeeded in generating iNKT cell cloned Trav11 - Traj18 +/+ mice from iPSC-iNKT cells through germline transmission and breeding with WT B6 mice ( 19 ). The absolute numbers and percentages of α-GalCer/CD1d dimer + TCRβ + cells in the thymus and periphery of Trav11 - Traj18 +/+ mice were elevated by 10–20-fold compared to B6 mice and 10–20-fold compared to B6 mice and by 3–10-fold compared to iNKT-TCRα transgenic mice due to the bypass of TCRα rearrangement at the double-negative (DN) stage.…”

“…They lacked γδ T cells due to the deletion of the δ locus and had reduced numbers of αβ T cells while NK, B, and DC numbers were normal. However, the surface phenotype of α-GalCer/CD1d dimer + TCRβ + cells in Trav11 - Traj18 +/+ mice was different from that in WT B6 mice; there was a partial reduction of CD44 + cells and changes in the CD4 + :NK1.1 + ratio ( 19 ). We think this is due to the shortage of CD1d molecules in the face of an excess number of α-GalCer/CD1d dimer + TCRβ + cells because the surface phenotype of the iNKT cells changed into the mature phenotype as seen in WT B6 when these cells were sorted and transferred into Traj18 −/− mice ( 58 ).…”

“…Moreover, iPS cell lines obtained by reprogramming of mature iNKT cells (iNKT-iPSC) from C57BL/6 (B6) mice preferentially generate iNKT cells but no conventional αβ T or γδ T cells, NK cells, DCs or B cells in vitro ( 18 ). Furthermore, mice generated from the iNKT-iPSC had a much larger number of iNKT-like cells ( 19 ) compared to mice with a rearranged Vα14-Jα18 transgene ( 16 ). It is therefore important to compare the development and function of iNKT cells and their subtypes that differentiate in vivo in these iNKT cell overexpressed mice.…”

Rapid and Efficient Generation of Transgene-Free iPSC from a Small Volume of Cryopreserved Blood

“…T-lymphocytes are another broadly used blood-derived cell source for reprogramming, even though cells need to be activated before reprogramming induction and often have much lower reprogramming efficiency. However, just like B-lymphocytes, T-lymphocytes harbor a set of gene rearrangements that may result in skewed development of T cells and perhaps other lineages [ 11 ], and even worse, could develop T cell lymphomas spontaneously [ 12 ]. Alternatively, expanded erythroblasts are a promising source for reprogramming using lentivirus, Sendai virus, or Epsterin-Barr virus (EBV)-based vector approaches [ 13 – 15 ].…”