Kluin-Nelemans J C, Kester M G, Melenhorst J J, Landegent J E, van de Corput L, Willemze R, Falkenburg J H
Department of Hematology, Leiden University Hospital, The Netherlands.
Blood. 1996 May 1;87(9):3795-802.
Hairy cell leukemia (HCL) is characterized by a severe T-cell-mediated immune deficiency. At the same time, spontaneous T-cell activation is noted when splenic T cells are studied in vivo and in vitro. Therefore, we searched for oligoclonal T-cell populations in the blood and spleens of 25 patients with HCL using a T-cell receptor gamma-polymerase chain reaction (TCR gamma-PCR). Subsequently, in 6 patients, the CDR3 length and conformation from 22 different TCRBV subfamilies were analyzed after PCR amplification of cDNA using TCRBV subfamily-specific primers. T cells from 15 of 25 HCL patients showed clonal excess by the TCR gamma-PCR. In fluorescence-activated cell sorted T-cell subsets, more clonal bands were observed than in the unseparated T cells, with most of these in CD8+ subsets, but also in CD4+, CD3+ gamma/delta+, and a double-negative CD3+ alpha/beta+ subset. In other B-cell malignancies, 6 of 16 samples showed oligoclonal T cells, whereas only 2 of 18 normal spleen and blood samples showed abnormal bands. Analysis of the TCRBV subfamilies disclosed in all 6 HCL patients a markedly abnormal pattern, with many clonal bands in 5 to 15 subfamilies, and absent or abnormal weak patterns in another 1 to 8 subfamilies. In comparison, 6 normal samples (2 spleens and 4 blood samples) showed in only 1 blood donor 1 clonal band. Two patients with active HCL but without infections or treatment were tested several times during the course of the disease and showed a complete identical skewed T-cell repertoire with the same oligoclonal T-cell populations. In conclusion, T cells in the blood and spleen of HCL patients show impressive abnormalities with many oligoclonal T-cell populations and a very restricted and skewed TCRBV repertoire.
毛细胞白血病（HCL）的特征是严重的T细胞介导的免疫缺陷。同时，在体内和体外研究脾T细胞时，可观察到自发的T细胞激活。因此，我们使用T细胞受体γ-聚合酶链反应（TCRγ-PCR）在25例HCL患者的血液和脾脏中寻找寡克隆T细胞群体。随后，在6例患者中，使用TCRBV亚家族特异性引物对cDNA进行PCR扩增后，分析了来自22个不同TCRBV亚家族的CDR3长度和构象。25例HCL患者中有15例的T细胞通过TCRγ-PCR显示克隆性过剩。在荧光激活细胞分选的T细胞亚群中，观察到的克隆条带比未分离的T细胞更多，其中大多数在CD8 +亚群中，但也在CD4 +、CD + γ/δ +和双阴性CD3 + α/β +亚群中。在其他B细胞恶性肿瘤中，16个样本中有6个显示寡克隆T细胞，而18个正常脾脏和血液样本中只有2个显示异常条带。对TCRBV亚家族的分析显示，所有6例HCL患者均呈现明显异常的模式，5至15个亚家族中有许多克隆条带，另外1至8个亚家族中则没有或呈现异常的弱模式。相比之下，6个正常样本（2个脾脏和4个血液样本）仅在1名献血者中显示1条克隆条带。两名活动性HCL患者在疾病过程中未感染或未接受治疗，多次检测显示具有完全相同的偏斜T细胞库和相同的寡克隆T细胞群体。总之，HCL患者血液和脾脏中的T细胞显示出令人印象深刻的异常，有许多寡克隆T细胞群体以及非常受限和偏斜的TCRBV库。
