Table 2. Analysis of engrafted mice... | American Society of Hematology

Table 2.

Analysis of engrafted mice

Experiment^*	Nature	Injected cells per animal			Analysis of engraftment
Experiment^*	Nature	% EGFP	Cell no. × 10⁶ before^†transduction	Cell no. × 10⁶ after^‡transduction	% CD45	% EGFP/ CD45	% 19⁺	% 33⁺	% EGFP/ CD19⁺	% EGFP/ CD33⁺	% CD19/ CD45	% CD33/ CD45
Control group
B	EGFP	58.60	0.25	1	3.23	0.96	2.7	1.2	0.97	0.43	83.59	37.15
D	EGFP	51	0.75	1.2	60.9	9.9	44.5	19.4	6.32	2.58	73.07	31.86
D	EGFP	51	0.75	1.2	7.02	3.83	2.93	1.51	1.63	0.91	41.74	21.51
D	EGFP	50	0.75	0.5	8.42	1.1	5.49	2.22	0.73	0.5	65.20	26.37
C	EGFP⁻	0	0.2	0.5	54.8	0	25.2	17			49.92	33.68
C	EGFP⁻	0	0.2	0.5	50.13	0	28.3	13			56.45	25.93
E	EGFP⁻	0	0.25	0.5	20.71	0	15.5	3.14			74.84	15.16
Test group
D	Ik6	18.80	0.45	2	5.32	1.61	2.06	1.01	0.79	0.29	38.72	18.98
D	Ik6	18.80	0.45	2	1.27	0.1	0.74	0.4	0	0.1	58.27	31.50
E	Ik6	15	0.1	1	10.65	0.84	7.34	0.69	0.72	0	68.92	6.48
E	Ik6	15	0.1	1	51.36	9.38	32.46	10.28	6.14	2.21	63.20	20.02
E	Ik6	15	0.1	1	11.46	6.21	8.23	1.81	5.1	0.1	71.82	15.79
F	Ik6	21	0.35	0.8	2.8	0.1	1.15	1.04	0	0.1	41.07	37.14
A^2-153	Ik6	16.3	0.09	0.5	11	0.7	4.59	6.07	0.28	0.44	41.73	55.18
A^2-153	Ik6	16.3	0.09	0.5	19.6	0.36	7.39	13.26	0	0.4	37.70	67.65
B^2-153	Ik6	18.50	0.15	0.4	11.13	0.94	3.05	6.1	0.1	0.8	27.40	54.81
D^2-153	Ik6	18.80	0.45	2	4.13	0.23	1.15	2.97	0.1	0.1	27.85	71.91

Experiment^*	Nature	Injected cells per animal			Analysis of engraftment
Experiment^*	Nature	% EGFP	Cell no. × 10⁶ before^†transduction	Cell no. × 10⁶ after^‡transduction	% CD45	% EGFP/ CD45	% 19⁺	% 33⁺	% EGFP/ CD19⁺	% EGFP/ CD33⁺	% CD19/ CD45	% CD33/ CD45
Control group
B	EGFP	58.60	0.25	1	3.23	0.96	2.7	1.2	0.97	0.43	83.59	37.15
D	EGFP	51	0.75	1.2	60.9	9.9	44.5	19.4	6.32	2.58	73.07	31.86
D	EGFP	51	0.75	1.2	7.02	3.83	2.93	1.51	1.63	0.91	41.74	21.51
D	EGFP	50	0.75	0.5	8.42	1.1	5.49	2.22	0.73	0.5	65.20	26.37
C	EGFP⁻	0	0.2	0.5	54.8	0	25.2	17			49.92	33.68
C	EGFP⁻	0	0.2	0.5	50.13	0	28.3	13			56.45	25.93
E	EGFP⁻	0	0.25	0.5	20.71	0	15.5	3.14			74.84	15.16
Test group
D	Ik6	18.80	0.45	2	5.32	1.61	2.06	1.01	0.79	0.29	38.72	18.98
D	Ik6	18.80	0.45	2	1.27	0.1	0.74	0.4	0	0.1	58.27	31.50
E	Ik6	15	0.1	1	10.65	0.84	7.34	0.69	0.72	0	68.92	6.48
E	Ik6	15	0.1	1	51.36	9.38	32.46	10.28	6.14	2.21	63.20	20.02
E	Ik6	15	0.1	1	11.46	6.21	8.23	1.81	5.1	0.1	71.82	15.79
F	Ik6	21	0.35	0.8	2.8	0.1	1.15	1.04	0	0.1	41.07	37.14
A^2-153	Ik6	16.3	0.09	0.5	11	0.7	4.59	6.07	0.28	0.44	41.73	55.18
A^2-153	Ik6	16.3	0.09	0.5	19.6	0.36	7.39	13.26	0	0.4	37.70	67.65
B^2-153	Ik6	18.50	0.15	0.4	11.13	0.94	3.05	6.1	0.1	0.8	27.40	54.81
D^2-153	Ik6	18.80	0.45	2	4.13	0.23	1.15	2.97	0.1	0.1	27.85	71.91

In the control group, 4 mice injected with cells submitted to retroviral transduction with the control LZRS-EGFP retroviral vector and 3 other animals (EGFP⁻) injected with the EGFP⁻ fraction after the retroviral transduction process were analysed. In the test group, 2 different patterns of hematopoietic reconstitution can be distinguished: 6 animals had more CD19⁺ cells than CD33⁺ cells, as usually observed in NOD-SCID mice transplanted with uncultured, untransduced CD34⁺ cells, or in the control group; 4 animals (

F2-153

) had more CD33⁺ cells than CD19⁺cells (Figure 5).

Different groups of animals, each group being engrafted with cells obtained from the same CB.

†

Equivalent number of fresh, uncultured cells.

‡

Actual number of cells that were injected into mice, following the transduction procedure, and ex vivo expansion of cell numbers.

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