高剂量化疗联合自体外周血造血干细胞移植时骨髓及外周血采集物中CD34+细胞黏附分子的表达

本研究的目的是应用免疫荧光直接三色标记和流式细胞术(FCM)，观测高剂量化疗(HDC)联合自体外周血造血干细胞移植(APBSCT)时不同阶段的骨髓及外周血CD34^ 细胞，表达CD54、CD49d和CD62L的情况，探讨不同来源的CD34^ 细胞表达黏附分子的差异及其临床意义。将动员前、干细胞采集后和移植结束而骨髓重建后的骨髓样本及每次外周血单个核细胞样本，以CD34-PE和CD45-PerCP标记的同时，分别加CD54-FITC、CD49d—FITC、CD62L—FITC直接三色荧光标记．应用FACS测定CD34^ 细胞及各黏附分子表达情况，并比较不同时段骨髓中各类黏附分子表达的差异，以及外周血造血干细胞采集物与动员前骨髓中各类黏附分子表达的差异。结果表明．动员前、采集后及移植重建后骨髓中CD34^ 细胞对CD54、CD49d和CD62L表达的变化无统计学差异；造血干细胞第1和第2次采集物之间CD34^ 细胞对CD54、CD49d和CD62L的表达变化也无统计学差异；而造血干细胞采集物中CD34^ 、CD49^ 细胞较之动员前骨髓中CD34^ CD49d^ 细胞明显减少(P=0．001)。结论：化疗联合粒细胞集落刺激因子(G-CSF)的动员方法，可使骨髓中CD34^ 细胞的CD49d表达下调，并使之动员而进入外周血，移植重建后骨髓中CD34^ 细胞的CD49d表达趋于正常，其进一步的临床意义有待更多的病例积累予以阐明。

Expression of Adhesion Molecules on CD34 + Cells of BM and PB Stem Cell Samples During High-Dose Chemotherapy Combined with Transplantation of Autologous PB Stem Cells

This study was aimed to investigate the expressions of adhesion molecules such as CD54, CD49d and CD62L by CD34 cells sampled from different stages of bone moarrow (BM) and peripheral blood (PB) before/after G CSF mobilization and after transplantation through the direct labeling with three colour immunofluorescence and flow cytometry, and to explore the differences in expression of adhesion molecules on CD34 cells from different origins and their clinical significance. Mononuclear cells collected from BM and PB before mobilization, after collection of stem cells and hematopoietic recostruction of BM at the end of transplantation were marked with CD54 FITC, CD49d FITC and CD62L FITC separately, as well as CD34 PE and CD45PerCE. 3 color fluorescene analysis was carried out by FACS. The expression differences of CD34 and adhesion molecules between BM and APBSC were compared. The results showed that expression differences of CD54, CD49d and cd62L on CD34 cells belore mobilization, after collection and reconstraction of transplantation were not statiscally significant, the difference of CD54, CD49d and CD62L on CD34 between 1st and 2nd collections of hematopoietic stem cells also were not statiscally significant. In the collected APBSC,the expression level of CD34 CD49d was significantly lower than those in BM before mobilization( P =0.001).It is concluded that the method of chemotherapy combined with G CSF mobilization can down regulate CD49d expression in BM CD34 cells,thus can mobilize and move theirs into peripheral blood.After the reconstitution by transplantation,the expression of CD49d on CD34 cells tends to normal,the clinical significance needs to be elucidated by accumulation of much more cases.