De novo human T cell development in NOD- scid IL2Rγ null mice was validated by 1) high levels of TCR excision circles, 2) complex TCRβ repertoire diversity, and 3) proliferative responses to PHA and streptococcal superantigen, streptococcal pyrogenic exotoxin. Coadministration of human Fc-IL7 fusion protein results in high percentages of human CD4 +CD8 + thymocytes as well human CD4 +CD8 − and CD4 −CD8 + peripheral blood and splenic T cells. Spleens from engrafted NOD- scid IL2Rγ null mice contain human Ig + B cells and lower numbers of human CD3 + T cells. These human cells include B cells, NK cells, myeloid cells, plasmacytoid dendritic cells, and HSC. Engraftment of NOD- scid IL2Rγ null mice with human HSC generate 6-fold higher percentages of human CD45 + cells in host bone marrow than with similarly treated NOD- scid mice. NOD- scid IL2Rγ null mice are deficient in mature lymphocytes and NK cells, survive beyond 16 mo of age, and even after sublethal irradiation resist lymphoma development. We report the development and characterization of a new genetic stock of IL-2R common γ-chain deficient NOD/LtSz- scid (NOD- scid IL2Rγ null) mice and document their ability to support human mobilized blood HSC engraftment and multilineage differentiation. To overcome this limitation, small animal models of human HSC engraftment have been used. This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.Ethical considerations constrain the in vivo study of human hemopoietic stem cells (HSC).

Further studies will be required to determine the relationship between this immune reaction and the vision loss frequently experienced by patients with optic disk hemorrhage. These findings are congruent with those obtained in our mouse-to-mouse transfers and indicate that, at least in this group of patients, an immune reaction directed against RGC develops when transferred to mice. Immunohistochemical evaluation of recipient eyes indicated that human lymphocytes occasionally extravasate and can be found in the mouse retinal ganglion cell layer. In contrast, transfer of PBMC from glaucoma patients resulted in a marked decrease of RGC density (1436☓10 RGC/mm 2, p=0.0003). Transfer of PBMC from non-glaucomatous patients did not decrease RGC density per mm 2 when compared to controls that also received microbead injection, but no PBMC (2427±498 RGC/mm 2 vs. Microbead occlusion resulted in mild elevation of IOP in all eyes. One animal displayed signs of host vs graft disease and was excluded from the study. The transfer of human PBMC to NSG mice was generally well tolerated. Additionally histochemical and immunohistochemical methods were used to evaluate retinal integrity and the presence of human lymphocytes in the mouse retina. Fifty days after transfer eyes were harvested and the density of gamma synuclein positive retinal ganglion cells (RGC) was determined in retinal flat mounts. Twenty days after transfer all eyes received one microbead injection to create a mild IOP challenge. Peripheral blood mononuclear cells (PBMC) were isolated by gradient centrifugation and transferred into immune deficient NOD/scid gamma (NSG) mice. Samples were also obtained from healthy age matched controls (N=10).

This study was conducted to evaluate whether a cellular immune reaction is also evoked by the mononuclear cells obtained from human glaucoma patients.īlood samples were obtained from primary open angle glaucoma patients who, despite normal IOPs, presented with optic disk hemorrhage - a clinical indication of disease progression (N=8). We previously demonstrated that adoptive transfer of CD3+ T-cells from glaucomatous to normal mice results in RGC loss in the recipient animals despite absence of elevated intraocular pressure (IOP).

Nod scid gamma mice