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. 2003 May;162(5):1629-38.
doi: 10.1016/s0002-9440(10)64297-6.

Preferential expansion of Vgamma9-JgammaP/Vdelta2-Jdelta3 gammadelta T cells in nasal T-cell lymphoma and chronic active Epstein-Barr virus infection

Michiko K Oyoshi  1 , Hiroshi Nagata, Nobuhiro Kimura, Yu Zhang, Ayako Demachi, Toshiro Hara, Hirokazu Kanegane, Yoshinobu Matsuo, Tomohiro Yamaguchi, Tomohiro Morio, Atsuyoshi Hirano, Norio Shimizu, Kohtaro Yamamoto
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Preferential expansion of Vgamma9-JgammaP/Vdelta2-Jdelta3 gammadelta T cells in nasal T-cell lymphoma and chronic active Epstein-Barr virus infection

Michiko K Oyoshi et al. Am J Pathol. 2003 May.

Abstract

We recently established an Epstein-Barr virus (EBV)-positive gammadelta T-cell line from a nasal T/natural killer (NK)-cell lymphoma (Nagata H, Konno A, Kimura N, Zhang Y, Kimura M, Demachi A, Sekine T, Yamamoto K, Shimizu N: Characterization of novel natural killer (NK)-cell and gammadelta T-cell lines established from primary lesions of nasal T/NK-cell lymphomas associated with the Epstein-Barr virus. Blood 2001, 97:708-713). Subsequently, we established two novel EBV-positive gammadelta T-cell lines from the peripheral blood of patients with chronic active EBV infection. Analysis of the terminal repeat of EBV showed that the three cell lines consisted of monoclonal populations, and flow cytometry showed that they had a common phenotype of gammadelta T cells: CD3(+) CD4(-) CD8(-) CD16(-) CD19(-) CD56(+) CD57(-) HLA-DR(+) T-cell receptor (TCR) alphabeta(-) TCR gammadelta(+). Analysis for the expression of TCR by flow cytometry showed that all three cell lines were Vgamma9(+)/Vdelta2(+), but negative for VgammaI, Vdelta1, or Vdelta3 TCR. Southern blot analysis for TCR genes showed that the three cell lines had a common rearrangement of Vgamma9-JgammaP and Jdelta3 genes. Polymerase chain reaction and sequence analysis of the junction between Vdelta and Jdelta genes revealed that the Jdelta3 genes were rearranged with the Vdelta2 genes. In contrast, none of the EBV-negative gammadelta T-cell lines, Molt-14, Peer, or Loucy, which were analyzed for controls, had Vgamma9 or Vdelta2 TCR, or a rearrangement of Jdelta3 genes. These results indicated that Vgamma9-JgammaP/Vdelta2-Jdelta3(+) gammadelta T cells were preferentially affected by EBV and expanded in patients with nasal gammadelta T-cell lymphoma and chronic active EBV infection. Jdelta3(+) gammadelta T cells are known to be a very minor population in gammadelta T cells of peripheral blood, whereas Vgamma9-JgammaP/Vdelta2-Jdelta1(+) cells are the major population. The close association of EBV with this particular gammadelta T-cell population may provide a key to the etiology of EBV-positive lymphoproliferative diseases.

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Figures

Figure 1.
Figure 1.
Schematic organization of the human TCRδ and TCRγ loci. The TCRδ locus is located entirely within the TCRα locus between the TCRVα and the TCRJα gene segments. TCRVγ genes are divided into four subfamilies (I to IV) based on sequence similarity. Each Vγ gene is assigned to its subgroup as indicated. The solid boxes are gene segments used in SNT cell lines.
Figure 2.
Figure 2.
Clonality of cells assessed using EBV terminal repeats. Southern blot analysis of the number of EBV-TRs as evidence for monoclonal expansion of EBV-positive cells in SNT cell lines. Lane 1, SNT-8; lane 2, SNT-13, and lane 3, SNT-15 cells. B95-8 cells (B) and Raji cells (R) were used as controls for cells with productive EBV expansion and monoclonal EBV replication, respectively.
Figure 3.
Figure 3.
Southern blot analysis for TCR genes. Shown are the hybridizations with the Jγ1 probe (A), the Jδ1 probe (B), and the Jδ3 probe (C). Southern blot analysis shows rearrangement of the Jγ1, Jδ1, and Jδ3 genes in all SNT cell lines. Human placental DNA was used as a germline configuration (c); 8, SNT-8; 13, SNT-13; 15, SNT-15 cells.
Figure 4.
Figure 4.
PCR analysis for the junction of Vδ and Jδ genes. PCR analysis shows the presence of the Vδ2-Jδ3 TCR gene rearrangement in the three SNT cell lines. Only the combination of a L-Vδ2 sense primer and a Jδ3 anti-sense primer amplified ∼500-bp PCR products identified as Vδ2-Jδ3. Lane 1, SNT-8; lane 2, SNT-13; lane 3, SNT-15 cells. No signal was detected in EBV-negative γδ T-cell lines; lane 4, Molt-14; lane 5, Peer; lane 6, Loucy.
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