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. 1998 Aug 18;95(17):10117–10122. doi: 10.1073/pnas.95.17.10117

Table 2.

Molecular analysis of single H/RS cells.

Patient Issue Exp. Primer set Cells positive PCR products Rearrangements
Repeated Unique
6104 Lymph node 1 VH, Vκ FRI  3/18 3  3 Vκ 1
Lymph node 2 VH, Vκ FRI  5/12 5  4 Vκ 1 1 Vκ 3
Cytospin 1 VH, Vκ FRI  5/16 6  5 Vκ 1 1 VH3
6444 Lymph node 1 VH, Vκ FRI 0/6
VH, Vκ FRI 1/6 1 1 VH3 6
VHL  4/17 4  4 VH3
Cytospin 1 VHL 10/18 10 10 VH3
Controls
6104 Lymph node 1 VH, Vκ FRI 1/3 B cells 1 1 VH4
0/8 neg. controls
Lymph node 2 VH, Vκ FRI 3/3 B cells* 6 All 6
0/5 neg. controls
Cytospin 1 VH, Vκ FRI 2/10 CD30 neg. 2 1 Vκ1, 1 VH3
0/4 buffer
6444 Lymph node 1 VH, Vκ FRI 0/1 B cells
0/2 neg. controls
VH, Vλ FRI 1/2 B cells 1 1 VH3
0/3 neg. controls
VHL 1/4 B cells 2 1 VH3, 1 VH4
0/8 neg. controls
Cytospin 1 VHL 0/12 neg. controls

Three different primer sets were used, as indicated: (i) six VH and four Vκ family-specific primers hybridizing to sequences within FRI (VH, VκFRI), (ii) The same six VH primers together with eight Vλ family-specific FRI primers (VH, Vλ FRI), and (iii) five VH leader family-specific primers (VH). Controls consisted of B and T cells micromanipulated on the same day from neighboring sections or independent cytospins. In one experiment, CD30-negative cells were micromanipulated as control from the same cytospin that was used for the isolation of HRS cells. Neg. controls consist of micromanipulated T cells and aliquots of the buffer covering the sections or cytospins. Repeated rearrangements indicates clonally related rearrangements; unique rearrangements are rearrangements obtained only once. The three B cell samples of the second experiment of case 6104 each contained two B cells.