非霍奇金淋巴瘤患者免疫球蛋白及T细胞受体基因重排研究

Study of immunoglobulin and T-cell receptor gene rearrangements in patients with non-Hodgkin＇ slymphoma

目的 研究非霍奇金淋巴瘤（NHL）患者骨髓或外周血免疫球蛋白（Ig）/T细胞受体（TCR）基因单克隆重排特点及其临床意义.方法 以BIOMED-2引物系统及多重PCR方法对139例NHL患者骨髓或外周血标本进行Ig及TCR基因重排检测.结果 在B系NHL（B-NHL）中,82例慢性淋巴细胞白血病（CLL）患者中有70例（85.4％）检出Ig基因单克隆重排,其中IgH、IgK、IgL单克隆重排阳性率分别为46.3％（38例）、62.2％（51例）和1.2％（1例）.其他类型的B-NHL患者有39.4％（33例中有13例）检出Ig基因单克隆重排,其中IgH和IgK单克隆重排阳性率分别为33.3％（11例）和39.4％（13例）,未检测到IgL基因单克隆重排.T系NHL（T-NHL）患者中有50.0％（24例中有12例）检出TCR基因单克隆重排,其中TCRB和TCRG单克隆重排阳性率分别为8.3％（2例）和45.8％（11例）,TCRB和TCRG双重基因重排阳性率为4.2％（1例）,未检测到TCRD单克隆重排.11例早期（Ann Arbor Ⅰ、Ⅱ期）与46例晚期（Ⅲ、Ⅳ期）患者的Ig/TCR基因单克隆重排阳性率分别为36.4％和45.6％；10例惰性患者和47例侵袭性患者的Ig/TCR基因单克隆重排阳性率分别为40.0％和44.7％,差异均无统计学意义（P值均＞0.05）.除外CLL的57例NHL患者骨髓涂片检测骨髓侵犯阳性率为12.3％,明显低于骨髓基因重排检测阳性率（43.9％）,两者差异有统计学意义（P＜0.05）.敏感性试验表明多重PCR检测恶性克隆的敏感性为3.12％～6.25％.结论 NHL的临床分期和恶性程度不同,其Ig/TCR基因单克隆重排阳性率有差异,但未能证实其相关性.联合应用BIOMED-2多重引物可提高PCR检测骨髓和（或）外周血Ig/TCR基因单克隆重排的敏感性.多重PCR对NHL患者骨髓侵犯检测的敏感性优于骨髓涂片检查。

Objective To investigate immunoglobulin （Ig） and T cell receptor （TCR） gene rearrangements in bone marrow or peripheral blood of patients with non-Hodgkin＇ s lymphoma（ NHL）, and to explore the potential clinical significance. Methods The Ig/TCR gene rearrangements in bone marrow or peripheral blood of 139 NHL patients were analyzed by using BIOMED-2 multiple primers system and Muhiplex PCR assay. Results Ig clonality was detected in 85.4% （70/82） of chronic lymphocytic leukemia （ CLL）, including 46.3% （38/82） IgH rearrangement, 62.2% （51/82） IgK rearrangement and 1.2% （1/82） IgL rearrangement, and in 39.4% （13/33） of other categories of B-lineage NHL （ B-NHL）, including 33.3% （11/33） IgH and 39.4% （13/33） IgK rearrangements. TCR clonality was detected in 50.0% （12/24） of all definite T-lineage NHL （ T-NHL）, including 8.3% （ 2/24 ） TCRB and 45. 8% （ 11/24 ） TCRG, no TCRD was detected. The detection rate of gene rearrangements of NHL diversed in different clinical stages [ 36.4% in early stage （ Ann Arbor stage I and 11 ） and 45.6% in late stage （ m and Ⅳ ） 1 and in different degrees of malignancy （40.0% indolent NHL and 45.6% aggressive NHL）, but no obvious statistical significance was obtained（P 〉0.05）. The detection rate of bone marrow invasions of NHL （except CLL） with ex- aminations of bone marrow smear under the microscope was 12.3% （7/57）, much lower than the clonalitytesting （43.9%, 25/57） （P 〈 0.05）. Sensitivity test showed that the sensitivity of malignant clonality testing by multiplex PCR was 3.12% - 6.25%. Conclusions The detection rate of gene rearrangements diverses in different clinical stages and degrees of malignancy of NHL, but the correlation has not been proved in this researeh. The sensitivity of mutiplex PCR-based elonality testing is enhaneed with the combination of BIOMED-2 primers system. It is more sensitive than the morphological examinations of bone marrow smear in deteeting bone marrow invasions, and may provide a powerful strategy in the routine diagnosis and assessment after treatment.