Oligoclonal TCRBV Gene Usage in B-Cell Chronic Lymphocytic Leukemia: Major Perturbations Are Preferentially Seen Within the CD4 T-Cell Subset

Abstract

TCRBV (T-cell receptor B variable) gene usage and CDR3 size distribution were analyzed using reverse transcription polymerase chain reaction (RT-PCR) to assess the T-cell repertoire of 10 patients with B-cell chronic lymphocytic leukemia (B-CLL) and in nine age-matched healthy control donors. When the usage of each TCRBV gene within the CD8+ T cells of the patients was compared with that of the controls, no statistically significant difference was noted except for BV 6S1-3. In contrast, within the CD4+ T cells of the CLL patients, a statistically significant overexpression for four BV families (2, 3, 5S1, 6S1-3) was seen while an underrepresentation was noted for five BV families (10, 11, 15, 16, 19). Based on the criterion that a value of any BV higher than the mean + 3 standard deviation (SD) of healthy controls indicated an overexpression, individual patients were shown to overexpress several TCRBV genes compared with the controls. Analyses of the CDR3 length polymorphism showed a significantly higher degree of restriction within CD4+ and CD8+ T cells of the patients, as compared with the corresponding control T-cell population. There was a significant difference in the CDR3 size distribution pattern with a more polymorphic CDR3 length pattern in the age-matched controls as compared with CLL patients, suggesting different mechanisms driving the T cells towards a clonal/oligoclonal TCRBV usage in patients and controls, respectively. The results show major perturbations of T cells in CLL patients, more frequently seen in the CD4+ T-cell subset, indicating that nonmalignant CD4+ T cells may be involved in the pathogenesis of CLL, but also CD8+ T cells.

Cytogenetic and molecular characterization of a patient with simultaneous B‐cell chronic lymphocytic leukemia and peripheral T‐cell lymphoma

“…The erythropoietic hyperplasia of undetermined signi®cance could also be due to a common lesion in a premalignant precursor. Other explanations for the development of the PTCL could be the long survival time of B-CLL patients, the decreased``immune surveillance'' due to the immunode®ciency that accompanies B-CLL or the chronic stimulation of T-helper cells by the neoplastic B-CLL clone [14,15].…”

Dendritic cells in patients with non‐progressive B‐chronic lymphocytic leukaemia have a normal functional capability but abnormal cytokine pattern

“…Surface marker staining was performed as described previously (Rezvany et al, 1999). Intracellular staining was carried out according to a modified method of Mendes et al (2000).…”

Autologous T lymphocytes recognize the tumour-derived immunoglobulin VH-CDR3 region in patients with B-cell chronic lymphocytic leukaemia

“…Mapping of T-cell epitopes might increase the possibility of synthetically producing effective vaccines that express various T-cell epitopes, not only cytotoxic T lymphocytes (CTLs), but also CD4Th 1 and CD4Th 2 epitopes that are of possible importance in tumour rejection (Hung et al, 1998;Pardoll & Topalian, 1998). Vaccination of patients with B-cell malignancies using the complete idiotype have shown promising results (Bendandi et al, 1999;Mellstedt & O È sterborg, 1999). One patient with a B-cell lymphoma was immunized with the autologous VH-CDR3 peptide and CTLs were induced (Wen & Lim, 1998).…”

“…Total RNA was extracted from 1±2 Â 10 6 leukaemic B cells and CD4 1 , CD8 1 T cells, using RNAzol B (Tel-Test, Friendswood, TX, USA) based on the guanidine thiocyanate phenol-chloroform extraction method (Chomczynski & Sacchi, 1987). First-strand cDNA was synthesized as previously described (Rezvany et al, 1999).…”