Generating progenitor cells with in vivo reconstitution functions has been at the center of intense research to accelerate biomedical applications of embryonic stem cells (ESCs) for the treatment of debilitating genetic, traumatic, and degenerative conditions [54].
Recently, humanized mice expressing HLA class I molecules have been generated but showed little improvement in human T cell reconstitution and function of T and B cells.
We have also used this approach to examine GPCR homodimerization by demonstrating reconstitution of function only upon co-expression of pairs of GPCR G-protein fusions that are individually inactive [ 30, 63].
It deserves mentioning, however, that these models have limitations, such as the relatively short-term maintenance of chimerism and the poor reconstitution and function of T, B, and natural killer lymphocytes.
For example, while HLA-A2 transgenic mice elicit a slight improvement in human T cell reconstitution and function of T and B cells, HLA-DR4 or HLA-DR5 transgenic mice had significantly increased human cell reconstitution and better immune responses, including Ig class switching and elevated human IgG responses [ 51– 51].
As reconstitution of MLKL function in MLKL KO cells can be used as an assay to evaluate the functions of different MLKL domains, we constructed vectors to express C-terminal Flag-tagged full-length, kinase domain, ND and N-terminal 10-amino-acid deletion (MLKL(11-464)) of murine MLKL and expressed each of them at comparable levels in MLKL KO L929 cells.
By reconstitution of MLKL function in MLKL-knockout cells, we showed that the N-terminus of MLKL is required for its function in necroptosis.
The goal of therapy was reconstitution of renal function by treatment of the underlying condition.
Ultimately, a molecular level understanding of mechanism will need to move beyond descriptive studies towards the biochemical reconstitution of Golgi function in vitro.
Therefore, to obtain better insight into the interplay between GVHD and GVL in transplanted patients, deeper understanding of the T-cell reconstitution and their function is essential.
