LACK OF RESTRICTED T-CELL RECEPTOR BETA-CHAIN VARIABLE REGION (V-BETA) USAGE OF REACTIVE T-LYMPHOCYTES IN HODGKINS-DISEASE

T-cell response against tumour-associated antigens is mediated by the TCR complex. To determine a possibly restricted TCR-V beta repertoire in reactive T-lymphocytes in Hodgkin's disease (HD), 20 cases (of whic h 10 were EBV-positive cases) were investigated using 14 monoclonal an tibodies (MoAbs) recognizing 11 different TCR-V beta region family pro ducts and Northern blot analysis with cDNA probes specific for mRNA tr anscripts of 11 V beta families that were not detectable by MoAbs. Fou r V beta families (V beta 5, V beta 6, V beta 8, V beta 19) were inves tigated using both immunohistochemistry (IHC) with anti-V beta MoAbs a nd Northern blot analysis. Immunohistochemical and Northern blot findi ngs were correlated with the detection of the Epstein-Barr virus (EBV) genome in Hodgkin's and Reed-Sternberg cells (H-RS). The non-neoplast ic lymphocytes in HD were predominantly of T-phenotype (CD3+). Most of these cells were TCR alpha beta+ (beta F1+) and only a few T-cells we re reactive for TCR-delta 1 antibody (TCR-gamma delta+). In the majori ty of cases helper/inducer T-cells (CD4+) outnumbered suppressor/cytot oxic T-cells (CD8+). Labelling of these samples with the panel of 14 a nti-V beta MoAbs showed that only a small percentage (0.2-5.5%) of bet a F1+ lymphocytes were positive with each of these MoAbs. The proporti on of these cells was comparable to that seen in normal tissues. Most TCR V beta+ cells were randomly distributed, but in virtually all case s occasional V beta+ cells pertaining to the various V beta families w ere seen in close contact to H-RS cells. Using total RNA extracted fro m malignant and normal tissues, no visible band was detected with the various V beta probes. As determined in the present study, the percent age of T-cells expressing a given V beta family must be greater than o r equal to 10% to be detected with Northern blot. Thus, the percentage of V beta+ cells expressing V beta families which were explored only with Northern blot were within the same range as those of the 11 diffe rent TCR-V beta region families assessed with IHC, i.e. 1-10% of lymph oid cells. The results of the present study show that in HD there is n o restricted T-cell V beta repertoire usage regardless of the detectio n of EBV. In addition, since the various V beta families are represent ed in T-cell subpopulations forming rosettes around H-RS cells, we con clude that the T-cells attracted by H-RS cells constitute a polyclonal population.