Meggetto F, al Saati T, Rubin B, Delsol G
Laboratoire d'Anatomie Pathologique, CHU Purpan, France.
Br J Haematol. 1994 Mar;86(3):524-32. doi: 10.1111/j.1365-2141.1994.tb04782.x.
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 which 10 were EBV-positive cases) were investigated using 14 monoclonal antibodies (MoAbs) recognizing 11 different TCR-V beta region family products and Northern blot analysis with cDNA probes specific for mRNA transcripts of 11 V beta families that were not detectable by MoAbs. Four V beta families (V beta 5, V beta 6, V beta 8, V beta 19) were investigated using both immunohistochemistry (IHC) with anti-V beta MoAbs and Northern blot analysis. Immunohistochemical and Northern blot findings were correlated with the detection of the Epstein-Barr virus (EBV) genome in Hodgkin's and Reed-Sternberg cells (H-RS). The non-neoplastic 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 were reactive for TCR-delta 1 antibody (TCR-gamma delta+). In the majority of cases helper/inducer T-cells (CD4+) outnumbered suppressor/cytotoxic T-cells (CD8+). Labelling of these samples with the panel of 14 anti-V beta MoAbs showed that only a small percentage (0.2-5.5%) of beta F1+ lymphocytes were positive with each of these MoAbs. The proportion of these cells was comparable to that seen in normal tissues. Most TCR V beta+ cells were randomly distributed, but in virtually all cases occasional V beta+ cells pertaining to the various V beta families were seen in close contact to H-RS cells. Using total RNA extracted from malignant and normal tissues, no visible band was detected with the various V beta probes. As determined in the present study, the percentage of T-cells expressing a given V beta family must be > or = 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 different TCR-V beta region families assessed with IHC, i.e. 1-10% of lymphoid cells. The results of the present study show that in HD there is no restricted T-cell V beta repertoire usage regardless of the detection of EBV. In addition, since the various V beta families are represented in T-cell subpopulations forming rosettes around H-RS cells, we conclude that the T-cells attracted by H-RS cells constitute a polyclonal population.
针对肿瘤相关抗原的T细胞反应由TCR复合体介导。为了确定霍奇金病（HD）反应性T淋巴细胞中可能受限的TCR-Vβ谱系，使用14种识别11种不同TCR-Vβ区域家族产物的单克隆抗体（MoAbs）以及用针对11种Vβ家族mRNA转录本的cDNA探针进行Northern印迹分析，对20例病例（其中10例为EBV阳性病例）进行了研究，这些mRNA转录本无法通过MoAbs检测到。使用抗Vβ MoAbs的免疫组织化学（IHC）和Northern印迹分析对4个Vβ家族（Vβ5、Vβ6、Vβ8、Vβ19）进行了研究。免疫组织化学和Northern印迹结果与霍奇金和里德-施特恩贝格细胞（H-RS）中爱泼斯坦-巴尔病毒（EBV）基因组的检测相关。HD中的非肿瘤性淋巴细胞主要为T表型（CD3+）。这些细胞大多数是TCR-αβ+（βF1+），只有少数T细胞对TCR-δ1抗体有反应（TCR-γδ+）。在大多数病例中，辅助/诱导性T细胞（CD4+）的数量超过抑制/细胞毒性T细胞（CD8+）。用这14种抗Vβ MoAbs对这些样本进行标记显示，只有一小部分（0.2 - 5.5%）的βF1+淋巴细胞对每种MoAbs呈阳性。这些细胞的比例与正常组织中的比例相当。大多数TCR Vβ+细胞随机分布，但几乎在所有病例中，偶尔可见属于不同Vβ家族的Vβ+细胞与H-RS细胞紧密接触。使用从恶性和正常组织中提取的总RNA，用各种Vβ探针未检测到可见条带。如本研究中所确定的，用Northern印迹检测到表达给定Vβ家族的T细胞百分比必须≥10%。因此，仅用Northern印迹探索的表达Vβ家族的Vβ+细胞百分比与用IHC评估的11个不同TCR-Vβ区域家族的百分比在同一范围内，即淋巴细胞的1 - 10%。本研究结果表明，在HD中，无论是否检测到EBV，T细胞Vβ谱系的使用都不受限。此外，由于不同的Vβ家族存在于围绕H-RS细胞形成花环的T细胞亚群中，我们得出结论，被H-RS细胞吸引的T细胞构成一个多克隆群体。
