Table 1.

Summary of Single-Cell Analysis of Two Cases for V Gene Rearrangements

Patient No. Experiment No.Cells Positive for V Rearrangements After PCR With Different Forward Primers PCR ProductsRearrangements
VκVHFRI VHFRII Clone- SpecificTotal Sequenced Repeated Unique
1  1a  ND  13/17  -150 ND  13  13  13 VH
 1b  ND   7/14  -150 7/14  7  7 VH1  
2  2′LN  0/13   0/13  -150 3/13  3  3  3 VH1  
 2"LN  0/7 -150 1/7  3/7  3  3  3 VH
 2"spleen  0/10  -150 1/10  2/10  2  2 VH1  
Control  1a-151 ND   6/8 T cells -150 ND  8  8   5 VH3,3 VH4  
 1b  ND   0/8 T cells  -150 0/8 T cells  
  ND   0/10 buffer  -150 0/10 buffer  
Control  2′LN  0/4 T cells   0/4 T cells -150 0/4 T cells  
 2"LN  0/6 T cells  -150 0/6 T cells  0/6 T cells  
 2"spleen  0/5 T cells -150 0/5 T cells  0/5 T cells      
Patient No. Experiment No.Cells Positive for V Rearrangements After PCR With Different Forward Primers PCR ProductsRearrangements
VκVHFRI VHFRII Clone- SpecificTotal Sequenced Repeated Unique
1  1a  ND  13/17  -150 ND  13  13  13 VH
 1b  ND   7/14  -150 7/14  7  7 VH1  
2  2′LN  0/13   0/13  -150 3/13  3  3  3 VH1  
 2"LN  0/7 -150 1/7  3/7  3  3  3 VH
 2"spleen  0/10  -150 1/10  2/10  2  2 VH1  
Control  1a-151 ND   6/8 T cells -150 ND  8  8   5 VH3,3 VH4  
 1b  ND   0/8 T cells  -150 0/8 T cells  
  ND   0/10 buffer  -150 0/10 buffer  
Control  2′LN  0/4 T cells   0/4 T cells -150 0/4 T cells  
 2"LN  0/6 T cells  -150 0/6 T cells  0/6 T cells  
 2"spleen  0/5 T cells -150 0/5 T cells  0/5 T cells      

H-RS cells were analyzed together with B and T cells coded in a blinded fashion from two patients with HD. The number of PCR products obtained and sequenced from H-RS and T cells for the first and the second experiment is indicated. Clonally independent VHgene rearrangements of various VH gene families were obtained from control B cells (not shown). Repeated rearrangements indicate clonally related rearrangements; unique rearrangements indicate rearrangements amplified once and not related to any other rearrangement. The sequences obtained from H-RS cells of both patients were deposited in the Genbank under the accession numbers AJ007389 and AJ0007390.

Abbreviations: LN, lymph node; ND, not determined.

F0-150

PCR was performed either with VHFRI or with VHFRII primers.

F0-151

In this experiment, a B-cell–specific antibody was eventually used for the “T-cell staining.”

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