Accumulation of Common Clonal T Cells in Multiple Lesions of Sarcoidosis

Table 1 Patient profiles

				BAL^b
Patient	Sex	Age	Serum ACE^a (6.0–21.0 IU/L)	Total Cell (×10⁵/ml)	%lymphocyte Count	CD4/8	Tissue Specimens Analyzed in this Study
1	F	60	35.2	2.5	43	7.23	PBMC^c, muscular sarcoid lesion (right crus)
2	F	58	21.0	1.5	4.5	5.00	PBMC, muscular sarcoid lesion (left crus)
3	F	39	9.8	1.0	28	3.47	PBMC, muscular sarcoid lesion (left crus)
4	M	35	16.9	10.0	12	7.09	PBMC, muscular sarcoid lesion (right femur), TBLB^e
5	M	70	12.1	2.0	21	11.14	PBMC, muscular sarcoid lesion (left crus), LN^d
6	M	36	5.2	2.0	30	1.58	PBMC, LN
7	M	68	15.0	1.0	13	6.59	PBMC, LN
8	F	69	6.0	1.5	19	6.07	PBMC, LN
9	F	47	19.3	1.5	15	4.46	PBMC, LN
10	M	55	16.8	1.5	19	6.81	PBMC, LN

^aACE, angiotensin converting enzyme; ^bBAL, bronchoalveolar lavage; ^cPBMC, peripheral blood mononuclear cells;
^dLN, lymph nodes; ^eTBLB, transbronchial lung biopsy. of a Cβ primer and each of 22 Vβ-specific primers (25). After 35 cycles of PCR (94°C for 1.5 min, 60°C for 2 min, 72°C for 3 min), the amplified DNA was diluted (1:20) in a denaturing solution (95% formamide, 10 mM EDTA, 0.1% bromophenol blue, and 0.1% xylene cyanol) and then was maintained at 90°C for 2 min. The diluted sample (2 µl) was electrophoresed in nondenaturing 5% polyacrylamide gels containing 10% glycerol, at 35 W of constant power, and at 25°C constant temperature for approximately 2 hr. The DNA was then transferred to Immobilon-S (Millipore Intertech, Bedford, MA), hybridized with a biotinylated Cβ probe (5′-biotin-A(AC)AA(GC)GTGTTCCCGAGGTCGCTGTGTT-3′) at 42°C for 12 hr, washed for 10 min at 55°C, and then visualized by subsequent incubations with streptavidin, biotinylated alkaline phosphatase, and a chemiluminescent substrate system (Phototope detection kit, New England Biolabs, Beverly, MA).

ISSN: 1528-3658