Table 3. Immunophenotyping of T Cells... | American Society of Hematology

Table 3.

Immunophenotyping of T Cells Derived From CD34⁺ Cells Transduced With the LNL6 or 9456 Ribozyme Retroviral Vectors

Day	Stage^3-150	LNL6			p9456t			Untransduced
		4⁺8⁺	4⁺	8⁺	4⁺8⁺	4⁺	8⁺	4⁺8⁺	4⁺	8⁺
15	Thymic culture	45	21	34	42	20	38	36	14	50
28	Expansion	34	18	48	46	24	30	30	28	42
42	Cell sorting	—	97	96	—	97	96	—	96	94
56	Expansion	—	93	96	—	96	93	—	94	95
70	G418	—	94	—	—	94	—	—	—	—
84	SIV	—	88	—	—	100	—	—	—	—
98	SIV	—	—	—	—	96	—	—	—	—

Day	Stage^3-150	LNL6			p9456t			Untransduced
		4⁺8⁺	4⁺	8⁺	4⁺8⁺	4⁺	8⁺	4⁺8⁺	4⁺	8⁺
15	Thymic culture	45	21	34	42	20	38	36	14	50
28	Expansion	34	18	48	46	24	30	30	28	42
42	Cell sorting	—	97	96	—	97	96	—	96	94
56	Expansion	—	93	96	—	96	93	—	94	95
70	G418	—	94	—	—	94	—	—	—	—
84	SIV	—	88	—	—	100	—	—	—	—
98	SIV	—	—	—	—	96	—	—	—	—

Expression of CD4 and CD8 was assessed on CD3⁺ T cells by three-color flow cytometric analysis. Results are expressed as a percentage of all CD3⁺ cells.

F3-150

CD34⁺ cells were transduced with the indicated retroviral vector, cultured on thymic stroma for 15 days, and expanded by stimulation with ConA, irradiated human PBMC, and IL-2; then CD4⁺ and CD8⁺ T cells were selected by cell sorting. These cells were then restimulated, cultured in the presence of G418 (200 μg/mL G418, active concentration), restimulated, and then infected with SIVmac239. Untransduced CD34⁺ cells were cultured in parallel up until the point of G418 selection.

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