T-cell receptor Vβ skewing frequently occurs in refractory cytopenia of childhood and is associated with an expansion of effector cytotoxic T cells: a prospective study by EWOG-MDS

A M Aalbers, M M van den Heuvel-Eibrink, I Baumann, H B Beverloo, G J Driessen, M Dworzak, A Fischer, G Göhring, H Hasle, F Locatelli, B De Moerloose, P Noellke, M Schmugge, J Stary, A Yoshimi, M Zecca, C M Zwaan, J J M van Dongen, R Pieters, C M Niemeyer, V H J van der Velden, A W Langerak, A M Aalbers, M M van den Heuvel-Eibrink, I Baumann, H B Beverloo, G J Driessen, M Dworzak, A Fischer, G Göhring, H Hasle, F Locatelli, B De Moerloose, P Noellke, M Schmugge, J Stary, A Yoshimi, M Zecca, C M Zwaan, J J M van Dongen, R Pieters, C M Niemeyer, V H J van der Velden, A W Langerak

Abstract

Immunosuppressive therapy (IST), consisting of antithymocyte globulin and cyclosporine A, is effective in refractory cytopenia of childhood (RCC), suggesting that, similar to low-grade myelodysplastic syndromes in adult patients, T lymphocytes are involved in suppressing hematopoiesis in a subset of RCC patients. However, the potential role of a T-cell-mediated pathophysiology in RCC remains poorly explored. In a cohort of 92 RCC patients, we prospectively assessed the frequency of T-cell receptor (TCR) β-chain variable (Vβ) domain skewing in bone marrow and peripheral blood by heteroduplex PCR, and analyzed T-cell subsets in peripheral blood by flow cytometry. TCRVβ skewing was present in 40% of RCC patients. TCRVβ skewing did not correlate with bone marrow cellularity, karyotype, transfusion history, HLA-DR15 or the presence of a PNH clone. In 28 patients treated with IST, TCRVβ skewing was not clearly related with treatment response. However, TCRVβ skewing did correlate with a disturbed CD4(+)/CD8(+) T-cell ratio, a reduction in naive CD8(+) T cells, an expansion of effector CD8(+) T cells and an increase in activated CD8(+) T cells (defined as HLA-DR(+), CD57(+) or CD56(+)). These data suggest that T lymphocytes contribute to RCC pathogenesis in a proportion of patients, and provide a rationale for treatment with IST in selected patients with RCC.

Trial registration: ClinicalTrials.gov NCT00499070 NCT00662090.

Figures

Figure 1
Figure 1
Frequency of skewing of individual TCRVβ families in bone marrow and peripheral blood of RCC and (v)SAA patients. (a) Frequency of skewing in RCC patients. (b) Frequency of skewing in (v)SAA patients.
Figure 2
Figure 2
Naive and effector CD4+ and CD8+ T cells in RCC patients with or without a skewed bone marrow or peripheral blood TCRVβ repertoire (Z-scores of relative distribution). (a) Lymphocytes within leukocytes. (b) CD3+/T cells within lymphocytes. (c) CD4+ within CD3+/T cells. (d) CD8+ within CD3+/T cells. (e) Naive CD4+ T cells. (f) Central memory CD4+ T cells. (g) Effector memory CD4+ T cells. (h) Terminally differentiated effector CD4+ T cells. (i) Naive CD8+ T cells. (j) Central memory CD8+ T cells. (k) Effector memory CD8+ T cells. (l) Terminally differentiated effector CD8+ T cells. Lines indicate medians. Z-scores <0 indicate a decrease and Z-scores >0 indicate an increase compared with age-matched controls. ns, nonsignificant; *P<0.05; **P<0.01; ***P<0.001.
Figure 3
Figure 3
Activation markers on CD8+ T cells in RCC patients with or without a skewed TCRVβ repertoire in bone marrow or peripheral blood (Z-scores of relative distribution). (a) CD56+CD8+ T cells. (b) CD57+CD8+ T cells. (c) HLA-DR+CD8+ T cells. (d) CD38+CD8+ T cells. Lines indicate medians. ns, nonsignificant; *P<0.05; ***P<0.001.

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