CAJ | 학술논문

目的通过建立我国范可尼贫血(Fanconi anemia,FA)患者原始B-淋巴细胞系进行FA分型。方法将我国3例FA患者外周血细胞用EB病毒活化法建立原始B-淋巴细胞系，采用丝裂霉素(MMC)生长抑制试验和细胞融合后MMC抑制试验的互补试验进行分型；利用单克隆抗体免疫沉淀法和Western blot法进行分型。结果成功建立了3个FA原始B-淋巴细胞系，其中2个细胞系仍然保持对MMC的敏感性，IC50均小于10nmol/L；用1例嵌合体患者细胞所建立的原始B-淋巴细胞系对MMC不敏感，IC50大于10mol/L；经MMC抑制试验的互补试验和Western blot检测，3例FA患者均为FA-A亚型。结论FA两种分型方法各有利弊，细胞融合后MMC生长抑制试验分型可靠，适用于所有未知FA亚型的患者，但该法依赖于建立MMC敏感的原始B-淋巴细胞系，无法用于FA-A嵌合体患者，方法耗时、耗力，出结果慢；而利用单克隆抗体免疫沉淀法和Westem blot法分型不依赖于建立MMC敏感的原始B-淋巴细胞系，可用于FA嵌合体患者，但仅适用于基因已知和已有特异性抗体的亚型，方法简单，出结果快。
목적통과건립아국범가니빈혈(Fanconi anemia,FA)환자원시B-림파세포계진행FA분형。방법장아국3례FA환자외주혈세포용EB병독활화법건립원시B-림파세포계，채용사렬매소(MMC)생장억제시험화세포융합후MMC억제시험적호보시험진행분형；이용단극륭항체면역침정법화Western blot법진행분형。결과성공건립료3개FA원시B-림파세포계，기중2개세포계잉연보지대MMC적민감성，IC50균소우10nmol/L；용1례감합체환자세포소건립적원시B-림파세포계대MMC불민감，IC50대우10mol/L；경MMC억제시험적호보시험화Western blot검측，3례FA환자균위FA-A아형。결론FA량충분형방법각유리폐，세포융합후MMC생장억제시험분형가고，괄용우소유미지FA아형적환자，단해법의뢰우건립MMC민감적원시B-림파세포계，무법용우FA-A감합체환자，방법모시、모력，출결과만；이이용단극륭항체면역침정법화Westem blot법분형불의뢰우건립MMC민감적원시B-림파세포계，가용우FA감합체환자，단부괄용우기인이지화이유특이성항체적아형，방법간단，출결과쾌。
Objective To Establish EBV-immortalized lymphoblast cell lines from Chinese Fanconi anemia(FA) patients. Methods The stablishment of lymphoblast cell lines was by EBV-immortalization and subtyping of the cell lines by cell fusion, mytomycin C (MMC) inhibition analysis and/or immunoprecipitation and Westemn blot. Result ①Three lymphoblast cell lines were established, two of them were sensitive to MMC with a IC50 ＜ 10 nmol/L. ②The subtypes of all the three patients belong to FA-A group. Conclusion Complementation analysis was suit-able for all FA patients with MMC-sensitive lymphoblastic cell lines, but this method is time consuming. Immuno-precipitation and Westemn blot are much faster for subtyping but could only be used for FA patients whose related genes had been cloned and specific antibodies been developed.