Hepatosplenic and Subcutaneous Panniculitis-Like γ/δ T Cell Lymphomas Are Derived from Different Vδ Subsets of γ/δ T Lymphocytes Grzegorz K. Przybylski,

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Hepatosplenic and Subcutaneous Panniculitis-Like γ/δ T Cell Lymphomas Are Derived from Different Vδ Subsets of γ/δ T Lymphocytes Grzegorz K. Przybylski, Hong Wu, William R. Macon, Janet Finan, Debra G.B. Leonard, Raymond E. Felgar, Joseph A. DiGiuseppe, Peter C. Nowell, Steven H. Swerdlow, Marshall E. Kadin, Mariusz A. Wasik, Kevin E. Salhany The Journal of Molecular Diagnostics Volume 2, Issue 1, Pages 11-19 (February 2000) DOI: 10.1016/S1525-1578(10)60610-1 Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Histology of representative cases of γ/δ HSTCL (A and B, Patient 5 inTable 2) and γ/δ SPTCL (C and D, Patient 12). A: Diffuse involvement of splenic parenchyma. B: Relatively monomorphic population of medium-size lymphocytes with round to oval nuclei, small nucleoli, and a moderate amount of pale cytoplasm. C: Lobular panniculitis-like pattern. D: Predominance of pleomorphic, small lymphocytes with irregular and hyperchromatic nuclei and scant cytoplasm. The Journal of Molecular Diagnostics 2000 2, 11-19DOI: (10.1016/S1525-1578(10)60610-1) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 FISH analysis of γ/δ HSTCL (Patient 2). A: CEP 8 probe to enumerate chromosome 8 centromeres showed the presence of trisomy 8 in 20% of the analyzed cells. B: Combination of CEP 7 probe (green staining) and LSI D7S486 probe (red staining) to detect simultaneously chromosome 7 centromere and band 7q31 on the chromosome's long arm showed the presence of isochrome 7 in 28% of the analyzed cells. A positive cell in metaphase is seen in the center of this photograph. Similar results showing the presence of both trisomy 8 and isochromosome 7 were obtained in patients 4 and 5 (not shown). The Journal of Molecular Diagnostics 2000 2, 11-19DOI: (10.1016/S1525-1578(10)60610-1) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 PCR analysis of Vδ gene usage in hepatosplenic γ/δ T cell lymphoma. A: Patient 2. B: Patient 3. Lane M, molecular weight marker; lanes 1–6, PCR products obtained with Vδ1-Vδ6 specific primers, respectively, and Jδ1 primer; lane C, negative control; lane 7, Dδ2Jδ1, lane 8, Vδ2Dδ3; lane 9, Dδ2Dδ3. The upper band of ∼600 bp represents RAG1 reference gene, bands of ∼400 bp in lanes A1 and B1 represent a Vδ1Jδ1 rearrangement, band of ∼140 bp in lane B7represents an incomplete Dδ2Jδ1 rearrangement. The Journal of Molecular Diagnostics 2000 2, 11-19DOI: (10.1016/S1525-1578(10)60610-1) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 PCR analysis of Vδ gene usage in archival paraffin-embedded samples of hepatosplenic γ/δ T cell lymphoma. Lane M, molecular weight marker; lanes 1–3, Patient 7; lanes 4–6, Patient 8; lane C, negative control. PCR was performed with the Jδ1 primer and the Vδ1b primer (lanes 1 and 4), the Vδ2b primer (lanes 2 and 5), and the Dδ2 primer- (lanes 3 and 6). The upper band of ∼220 bp represents RAG2 reference gene, bands of ∼120 bp in lanes 1 and 4 represent a Vδ1Jδ1 rearrangement, and the band of ∼140 bp in lane 6 represents an incomplete Dδ2Jδ1 rearrangement. The Journal of Molecular Diagnostics 2000 2, 11-19DOI: (10.1016/S1525-1578(10)60610-1) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 PCR analysis of Vδ gene usage in subcutaneous panniculitis-like γ/δ T cell lymphoma. A: Patient 14. B:Patient 15. Lane M, molecular weight marker; lanes 1–6, PCR products obtained with Vδ1-Vδ6 specific primers, respectively, and the Jδ1 primer; lane C, negative control; lane 7, Dδ2 and Jδ1 primers, lane 8, Vδ2 and Dδ3 primers; lane 9, Dδ2 and Dδ3 primers. The upper band of ∼600 bp represents RAG1 reference gene, bands of ∼400 bp in lanes A2 and B2 represent a Vδ2Jδ1 rearrangement, and the band of ∼140 bp in lane A7 represents an incomplete Dδ2Jδ1 rearrangement. The Journal of Molecular Diagnostics 2000 2, 11-19DOI: (10.1016/S1525-1578(10)60610-1) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions