Rapid generation of clinical-grade antiviral T cells: selection of suitable T-cell donors and GMP-compliant manufacturing of antiviral T cells

Table 1 Verification of CMV-specific T-cell frequencies in potential T-cell donors selected from the alloCELL registry

Donor	HLA-typing					*alloCELL*		Verification and detailed analysis of CMV-specific memory T-cell frequencies
	HLA-typing					TCR-pMHC interaction	EliSpot	Staining of T-cell subsets			TCR-pMHC interaction	CSA		EliSpot
	A	B	C	DRB1	DQB1	% A02pp65_M	spw	% CD3	% CD4	% CD8	% A02pp65_M	%OF	%TCF	spw
	A	B	C	DRB1	DQB1	[CD19⁻ CD3⁺CD8⁺]	[IFN-γ⁺]	[CD45⁺CD19⁻]	[CD3⁺]	[CD3⁺]	[CD19⁻ CD3⁺CD8⁺]	[CD3⁺IFN-γ⁺]	[CD3⁺IFN-γ⁺]	[IFN-γ⁺]
1	02:01 68:01	08:01 39:01	07:01 12:03	01:01 03:01	02:01 05:01	2.45	273	78.88	52.47	41.24	1.5	1.7	75.48	236
2	25:01 32:01	08:01 35:01	04:01 07:01	03:01 14:01	02:01 05:03	n.a.	162	59.65	69.53	26.61	n.a.	0.21	38.39	178.4
3	02:01 11:01	27:02 55:01	02:02 03:03	15:01 16:01	05:02 06:02	0.31	142	63.79	68.41	26.7	0.34	7.6	89.63	306.5

Three potential T-cell donors we selected according to their CMV-seropositivity and CMVpp65-specific T-cell frequencies from the allogeneic cell registry alloCELL. For verification, PBMCs were isolated, T-cell subsets were stained (% T cells [CD3, CD4 and CD8]) and CMVpp65-specific memory T cells were detected directly by pMHC pentamer staining using the A02pp65_M (donor 1 and 3, % A02pp65_M ⁺/CD3⁺CD8⁺ T cells) and after a short ex vivo stimulation with the CMVpp65_pp by IFN-γ-based CSA (% CD3⁺IFN-γ⁺ T cells (OF and TCF) and by IFN-γ EliSpot (spw). Because the donor was HLA-A*02:01 negative = n.a. (not applicable).

ISSN: 1479-5876