异基因外周血干细胞移植患者人类白细胞抗原B_(27)的表达

Expression of homologous leucocytic antigen B_(27) after allogeneic peripheral blood stem cell transplantation

目的:分析白血病患者行异基因外周血干细胞移植后人类白细胞抗原B27的表达特点,及其与慢性移植物抗宿主病的关系。方法:选择2003-09/2006-06在广西医科大学第一附属医院行异基因外周血干细胞移植术的45例白血病患者,按《血液病诊断和疗效标准》进行疾病的诊断和分型。全部患者均知情同意并签署知情同意书。男27例,女18例,22～51岁。其中急性髓细胞白血病13例、急性淋巴细胞白血病17例、慢性粒细胞白血病15例。供者均为白细胞抗原配型6个位点全相合的同胞兄妹。急性髓细胞白血病和慢性粒细胞白血病患者采用BU/CY方案:马利兰4mg/(kg·d)×4d,环磷酰胺60mg/(kg·d)×2d;急性淋巴细胞白血病患者预处理采用TBI/CY方案:全身照射5.0Gy/d×2d,环磷酰胺60mg/(kg·d)×2d。供者采用粒细胞集落刺激因子皮下注射动员干细胞。采用美国Baxter CS-3000plus血细胞分离机采集外周血干细胞,每位供者采集0,+1,+2d3次。回输单个核细胞、粒-单核系集落形成单位、CD34+细胞。采用流式细胞术动态检测45例白血病患者异基因外周血干细胞移植前后人类白细胞抗原B27表达的变化。结果:45例患者全部进入结果分析。①45例患者均获得造血重建,经荧光标记PCR短串联重复序列DNA位点分析,全部患者移植后血细胞的DNA与供者血细胞DNA完全一致。发生急性移植物抗宿主病4例,慢性移植物抗宿主病14例,出血性膀胱炎3例,肝静脉闭塞病4例。②移植后发生慢性移植物抗宿主病患者人类白细胞抗原B27表达水平明显低于无移植物抗宿主病患者[移植后180d:(30.13±10.24)%,(39.61±10.89)%;移植后峰值:(33.53±10.36)%,(45.76±12.09)%,P<0.05]。③移植前未缓解白血病患者人类白细胞抗原B27表达水平则明显高于移植前达到完全缓解患者[(27.39±10.31)%,(16.62±8.58)%,P=0.005],移植后疾病复发患者复发时人类白细胞抗原B27表达水平明显高于其移植后完全缓解时[(27.76±10.09)%,(15.69±6.98)%,P=0.015]。移植前未缓解和移植后复发患者人类白细胞抗原B27表达水平与肿瘤细胞占骨髓非红系细胞的百分比呈正相关关系(r=0.756,P=0.000)。结论:移植前未缓解和移植后复发白血病患者人类白细胞抗原B27表达与肿瘤细胞占骨髓非红系细胞的百分比呈正相关,人类白细胞抗原B27表达水平随着肿瘤细胞百分比的增高而升高。

AIM： To analyze the law of homologous leucocytic antigen （HLA）-B27 expression and its relation with chronic graft-versus-host disease （cGVHD） after allogeneic peripheral blood stem cell transplantation （Alio-PBSCT） in leukemia patients. METHODS： Totally 45 leukemia patients who had undergone Allo-PBSCT in First Affiliated Hospital, Guangxi Medical University between September 2003 and June 2006 were selected. Diagnosis and typing of disease was done according to Diagnosis and Curative Criteria of Hematonosis. All patients knew the trial and signed the informed consent, including 27 males and 18 females, aged from 22-51 years. There were 13 cases of acute myeloid leukemia （AML）, 17 cases of acute lymphoblastic leukemia （ALL） and＇15 cases of chronic myeloid leukemia （CML）. All donors were HLA-identical sibling. Conditioning regimens were BU/CY [busulfan 4 mg/kg per day×4 d,cyclophosphamide 60 mg/kg per day×2 d] of AML and CML, or TBI/CY [total body irradiation 5.0 Gy/dx2 d, cyclophosphamide 60 mg/kg per day×2 d] of ALL. Stem cells were mobilized with granulocyte colony-stimulating factor from HLA-idenUcal sibling donors. Three leukapheresis procedures （0, ＋1, ＋2 days） were performed by Baxter CS 3 000 plus blood cell separator to collect peripheral blood stem cells. Mononuclear cells, CFU-GM and CD34^＋ cells were transfused. Flow cytometry was used to detect the expression level of HLA-B27 in peripheral blood of 45 cases with leukemia after Allo-PBSCT. RESULTS： Totally 45 patients were involved in the result anlaysis. ①All 45 patients were successfully engrafted and short tandem repeats with fluorescent labeling polymerase chain reaction were analyzed, all patients with full donor chimeras. Acute GVHD occurred in 4 cases and cGVHD in 14 cases, hemorrhagic cystitis occurred in 3 cases and vertous occlusive disease in 4 cases.②The expressions of HLA-B27 ware markedly lower in the patients with cGVHD than that without cGVHD [180 days after Allo-PBSCT： （30.13±10.24）%, （39.61±10.89）%;peak value after Allo-PBSCT： （33.53±10.36）%, （45.76±12.09）%,P 〈 0.05].③ The level of HLA-B27 expression was obviously higher in non-remission before transplantation group than that in achieved complete remission before transplantation group [（27.39±10.31 ）%, （16.62±8.58）% ,P =0.005]. The expressions of HLA-B27 ware significantly higher in the patients with relapsed after transplantation than those with complete remission after transplantation [（27.76±10.09）%, （15.69±6.98）%,P=0.015]. There was a significantly positive correlation between the expressions of HLA-B27 and the percentage of leukemic blast cells in bone marrow （r=0.756, P =0.000）. CONCLUSION： There is a significantly positive correlation between the expression of HLA-B27 and the percentage of leukemic blast cells in bone marrow in the patients with non-remission before transplantation and relapsed after transplantation. The expression of HLA-B27 increases with the increase of percentage of leukemic blast cells in bone marrow.