Table 1.

Impact of AZA on engraftment, proliferation and immunomodulation. NSG mice were transplanted with 2 × 10⁷ human PBMC, treated or not with AZA, and FACS was performed on blood cells on day 14 or FACS and RT-qPCR were performed on spleen, blood and bone marrow cells on day 28 after sacrifice. Human chimerism was calculated as the ratio of human CD45⁺ cells (_huCD45⁺) vs. murine+human CD45⁺ cells. CFSE dilution of _huCD45⁺ cells was analyzed in spleen, 4 d after transplantation of 4 mice/condition with 2 × 10⁷ CFSE-loaded human PBMCs. Blood absolute counts were calculated based on white blood cell count performed with hematological cell counter (Sysmex) and _hu/muCD45⁺ cell frequency determined by FACS. Transcript levels measured by RT-qPCR were normalized on human housekeeping gene (TBP). Data show median values of 8–10 mice/condition with interquartile range (IQR).

		CTRL		AZA
Parameter	Method	Median	IQR	Median	IQR	p-value
Human chimerism (%):	FACS
Day 14, peripheral blood		10.3	8.86–12.94	26.91	22.84–37.84	0.0007
Day 28, peripheral blood		60.09	51.72–70.25	90.94	66.95–93.49	0.014
Day 28, spleen		83.09	78.73–87.33	66.5	57.50–83.67	0.1893
Day 28, bone marrow		3.25	1.05–9.09	10.04	8.32–18.79	0.041
KI67+ among huCD45+ (%):	FACS
Day 14, peripheral blood		38.15	33.20–47.38	20.3	14.65–24.80	< 0.0001
Day 28, spleen		20.8	14.28–24.35	43.9	36.20–48.45	< 0.0001
CFSEdim among huCD45+ (%):	FACS
Day 4, spleen		69	64.30–75.80	48.6	46–50.4	0.0039
Blood absolute count (103/µL):	Sysmex and FACS
Day 14, huCD45+		0.127	0.093–0.199	0.056	0.047–0.082	0.0093
Day 14, muCD45+		0.168	0.131–0.359	0.051	0.037–0.11	0.0012
Sacrifice day 28, spleen:
% CD4+CD25+HLA-DR+ T cells	FACS	1.935	1.52–4.218	12.4	8.47–14.15	< 0.0001
% CD8+CD25+HLA-DR+ T cells		3.095	2.683–3.67	4.88	4.03–7.125	0.017
% IL-2+ among CD4+ T cells		7.86	7.33–10.90	22.6	19.40–26.60	0.0003
IL2 relative expression	RT-qPCR	0.574	0.355–0.842	1.35	1.22–1.56	0.0002
% IFNγ+ among CD4+ T cells	FACS	67.4	66.90–73.50	60.5	57.40–65.50	0.0079
IFNG relative expression	RT-qPCR	1.281	0.944–1.7	0.36	0.213–0.777	0.0021
TBET relative expression		1.82	1.578–2.315	0.689	0.58–0.891	< 0.0001
GATA3 relative expression		1.515	1.3–1.935	0.819	0.623–1.29	0.0025
% IL-17+ among CD4+ T cells	FACS	1.81	1.74–3.01	2.02	1.855–2.37	0.873
RORC relative expression	RT-qPCR	1.403	1.107–2.111	2.111	1.384–3.685	0.1903
% GZMB+ among CD8+ T cells	FACS	70.85	68.63–78.55	46.7	43.1–55.6	0.0003
% PRF1+ among CD8+ T cells		69	63.23–76.25	47.2	39.6–51.4	0.0006
Serum cytokine day 28 (pg/mL)
IFNγ	Bio-Plex	1554	1,450–2,348	31.25	23.46–78.80	0.0002
TNF-α		47.33	20.24–61.47	5.03	2.27–5.48	0.0002
IL-2		1.76	1.32–3.43	3.11	2.29–3.61	0.1304