Table 1.

Deduced amino acid sequences of paired TCR α- and β-chains of microdissected T cells from muscle biopsy tissue

Patient	Chain and sequence					No. of cells with TCR identified by
	V		N(D)N	J		Universal primer set	Clone-specific primer
PM16488 (1,230 cells analyzed)
β-Chain	1.1	CASSV	GGL	YGYTF	1.2		64
α-Chains	2.2	CAM		SQGAQKLVF	54	6	20
	11.1	CAV	GS	NARLMF	31	1	1
	20.1	CL	LSS	NTGKLIF	37	1	1
	23.1	CAV	D	SGTYKYIF	40	1	1
IBM15551 (250 cells analyzed)
β-Chain	23.1	CASS	SDRE	NTEAFF	1.1		8
α-Chains	4.2	CI	ASV	YGNKLVF	47	1	1
	23.1	CA	VL	NAGNMLTF	39	1	1

In a first step, we detected TCR β-chain sequences of expanded clones by CDR3 spectratyping. The deduced amino acid sequences are shown. Then, biopsy sections were stained by appropriate anti-Vβ antibodies, and Vβ-positive cells were isolated by microdissection. We tested these cells for the expression of the correct, expanded TCR β-chains by single-cell PCR using clone-specific primers. We identified 64 cells from patient PM16488 and 8 cells from patient IBM15551 that had identical sequences as determined by CDR3 spectratyping and single-cell PCR. These β-chain-positive cells were then tested for α-chain expression by using a universal primer set that allows amplification of all possible TCR α-chains. The sequences of the α-chains and the numbers of cells identified are listed. Based on the sequences of these α-chains, all β-chain-positive cells were reexamined for α-chains by using clone-specific α-primers. In all cells in which we had identified an α-chain with the universal primer set, we confirmed expression of this α-chain with clone-specific primers. In patient PM16488, we identified the Vα2.2-Jα54 sequence in 14 additional cells.