Human CD8⁺/TCR⁻/CD56^dim/− facilitating Cells Enhance Engraftment of HSC In NOD/SCID/IL2rγ^null Recipients.

Abstract 3709

CD8⁺/TCR⁻ graft facilitating cells (FC) enhance engraftment of hematopoietic stem cells (HSC). They are distinct from HSC since they do not generate multilineage engraftment when infused alone, nor do they form colonies in vitro. The function of human FC remains to be defined. Here, we phenotypically characterized the human CD8⁺/TCR⁻ FC subpopulations and evaluated their function in vivo and in vitro. Human CD8⁺/TCR⁻ FC comprised 1.1% ± 0.27% of total G-CSF-mobilized peripheral blood mononuclear cells (PBMC). Approximately 55% of FC express CD56^dim/−, and the remaining population is CD56^bright. In the CD8⁺/TCR⁻/CD56^bright cell population, approximately 65% of cells express CD11c⁺ and 67% of cells express CD11b⁺. In CD8⁺/TCR⁻/CD56^dim/− cell population, the majority of cells express CD3ε⁺ (80%), 17% were CD11c⁺, 16% were CD19⁺, 14% were CD11b⁺, 11% were CD123⁺, and 30% were HLA-DR⁺. To evaluate whether human CD8⁺/TCR⁻/CD56^dim/− FC enhance engraftment of human HSC in vivo, we transplanted 100,000 HSC alone or plus 300,000 CD8⁺/TCR⁻/CD56^dim/− FC into NOD/SCID/IL2rγ^null (NSG) recipient mice conditioned with 325 cGy of total body irradiation. At 30 days after transplantation, 8 of 21 (38%) recipients of HSC alone engrafted. In contrast, 81% of recipients (n = 16) receiving HSC plus CD8⁺/TCR⁻/CD56^dim/− FC engrafted, and donor lymphocyte and monocyte chimerism in PB was 0.53% ± 0.16% and 3.93% ± 1.28%, respectively. At 6 months after transplantation, NSG recipients of HSC alone lost donor chimerism in PB and no donor cells were detected in spleen and BM. In contrast, NSG recipients of HSC + CD8⁺/TCR⁻CD56^dim/− FC exhibited durable donor chimerism in PB and showed significantly higher levels of donor chimerism in spleen (13%) and BM (9.43%) compared to recipients of HSC alone. To evaluate the function of CD8⁺/TCR⁻/CD56^dim/− FC in vitro, HSC were incubated with CD8⁺/TCR⁻CD56^dim/− FC for 18 hrs and then cultured in methylcellulose for 14 days in colony-forming cell assay. HSC plus CD8⁺/TCR⁻CD56^dim/− FC generated significantly more colonies compared with HSC alone (P = 0.0038), suggesting that human CD8⁺/TCR⁻/CD56^dim/− FC have a direct effect on the clonogenicity of HSC (Figure). In summary, our data indicate that (1) human CD8⁺/TCR⁻ FC are a heterogeneous cell population; (2) CD8⁺/TCR⁻/CD56^dim/− FC enhance engraftment of human HSC in vivo and promote HSC clonogenicity in vitro; and (3) recipients of CD8⁺/TCR⁻/CD56^dim/− FC maintain a durable donor chimerism in BM and spleen.

View largeDownload slide

View largeDownload slide

Close modal