【摘要】：目的:探討成人EB病毒相關(guān)淋巴細(xì)胞克隆性轉(zhuǎn)化的實(shí)驗(yàn)方法,為EB病毒相關(guān)淋巴細(xì)胞增殖病的早期診斷、分類、預(yù)測(cè)轉(zhuǎn)歸提供客觀的判斷指標(biāo)。方法:采集5例成人EB病毒相關(guān)的淋巴組織增殖性疾病(EBV+LPD)患者的外周血標(biāo)本,另采集4例成人傳染性單核細(xì)胞增多癥(IM)外周血標(biāo)本作為陰性對(duì)照,3例急性NK細(xì)胞白血病(ANKL)的外周血標(biāo)本作為陽性對(duì)照。應(yīng)用流式細(xì)胞術(shù)(FCM)檢測(cè)淋巴細(xì)胞免疫表型、RT-PCR檢測(cè)T細(xì)胞受體(TCR)基因重排,Southern blot方法鑒定EBV末端重復(fù)序列(TR)多態(tài)性,對(duì)患者外周淋巴細(xì)胞進(jìn)行了克隆性分析。結(jié)果:在5例EBV-LPD患者中,FCM僅檢測(cè)出1例TCRVβ表型呈克隆性;RT-PCR檢測(cè)TCRVβ時(shí)在IM患者均檢測(cè)出TCR受體基因重排陽性,在5例EBV-LPD患者中檢測(cè)出4例陽性;在5例EBV+LPD患者中1例檢測(cè)有單克隆條帶,2例檢測(cè)有寡克隆條帶。結(jié)論:流式細(xì)胞術(shù)表型分析的敏感性較差;TCRVβ重排不能區(qū)分反應(yīng)性克隆和轉(zhuǎn)化型克隆;而EB病毒末端重復(fù)序列克隆性鑒定的方法,能夠更客觀特異地反應(yīng)EB病毒相關(guān)淋巴細(xì)胞增殖的克隆性轉(zhuǎn)化,有助于改善疾病的早期診斷、分類及預(yù)測(cè)臨床轉(zhuǎn)歸。
[Abstract]:Objective: to investigate the experimental method of clonal transformation of adult EB virus-associated lymphocytes in order to provide an objective index for the early diagnosis, classification and prognosis of EB virus-associated lymphoproliferative disease. Methods: peripheral blood samples of 5 adult patients with EB virus-associated lymphoproliferative disease (EBV LPD) and 4 adult infectious mononucleosis (IM) peripheral blood samples were collected as negative control. Peripheral blood samples from 3 patients with acute NK cell leukemia (ANKL) were used as positive control. The lymphocyte immunophenotype was detected by flow cytometry (FCM), and the (TR) polymorphism at the end of EBV was identified by RT-PCR analysis of T cell receptor (TCR) gene rearrangement, Southern blot. The clonal analysis of peripheral lymphocytes was carried out. Results: in 5 cases of EBV-LPD, only 1 case of TCRV 尾 was detected in FCM, TCRV 尾 was detected in all cases of IM by RT-PCR, and 4 cases were positive in 5 cases of EBV-LPD. Monoclonal bands and oligoclonal bands were detected in 1 of 5 patients with EBV LPD. Conclusion: the sensitivity of phenotypic analysis by flow cytometry is poor, and TCRV 尾 rearrangement can not distinguish reactive clone from transformed clone. The clone identification method of terminal repeat sequence of EB virus can more objectively and specifically reflect the clonal transformation of lymphocyte proliferation associated with EB virus, which is helpful to improve the early diagnosis, classification and prediction of clinical outcome of EB virus.
【作者單位】： 北京大學(xué)深圳醫(yī)院血液內(nèi)科;
【分類號(hào)】：R55

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