背景:外周血干细胞移植成功的首要条件是干细胞的有效动员和采集,选择高效低毒的动员方案,掌握动员和采集时机与动员效果密切相关.目的:探讨米托蒽醌-大剂量阿糖胞苷方案化疗后,单用粒细胞集落刺激因子或粒细胞集落刺激因子与粒-巨噬细胞集落刺激因子合用对恶性血液病和实体瘤患者自体外周血干细胞的动员效果.设计:观察对比实验.单位:徐州医学院附属医院血液科.对象:选择1998-09/2006-12在徐州医学院附属医院血液科收治的42例恶性血液病和实体瘤患者,诊断符合国际白血病分型及世界卫生组织新分类标准.男25例,女17例,年龄7～54岁,平均29岁,体质量(52±18)kg.其中急性髓细胞白血病12例,急性淋巴细胞白血病6例,慢性粒细胞白血病慢性期1例,非霍奇金淋巴瘤15例,霍奇金淋巴瘤4例,多发性骨髓瘤2例,晚期乳癌2例.患者均经常规化疗达到或接近完全缓解,骨髓细胞学检查无肿瘤细胞浸润.心、肺、肝、肾等主要脏器功能正常.动员前化疗疗程平均8次,所有患者均对治疗项目知情同意.方法:患者均采用米托蒽醌10 mg/(m2·d)静脉滴注第2～3 d后,阿糖胞苷2 g/m2静脉滴注第1～2 d,1次/12 h.当白细胞计数下降至最低点开始回升时,20例患者使用粒细胞集落刺激因子5～7.5 μg/(kg·d),连用3～5 d,22例患者早6:00给予粒细胞集落刺激因子5～7.5 μg/(kg·d),晚6:00给予粒-巨噬细胞集落刺激因子5～7 μg/(kg·d).白细胞计数＞2.5×109 L-1,CD34+细胞≥1%时,用CS 3 000 plus血细胞分离机连续2 d采集自体外周血干细胞,检测CD34+细胞含量和T淋巴细胞亚群.①单个核细胞与FITC标记的CD34+、CD3和CD8单抗及与CD4PE标记的CD4单抗4 ℃混合30 min,采用流式细胞仪检测CD34+细胞和T细胞亚群,分析5×105个细胞,得出CD3、CD34+细胞含量及CD4/CD8比值.用甲基纤维素法测定粒-巨噬细胞集落形成单位.②观察术后相关不良反应.③针对不同类型疾病给予相应预处理36～48 h后回输自体外周血干细胞,并行单个核细胞计数及台盼蓝染色,解冻后检测粒-巨噬细胞集落形成单位和CD34+细胞.主要观察指标:①动员前后CD34+细胞和T细胞亚群变化.②术后相关不良反应.③自体外周血干细胞回输量(单个核细胞计数、粒-巨噬细胞集落形成单位和CD34+细胞数).结果:纳入患者42例,均进入结果分析.①动员前后CD34+细胞和T细胞亚群变化:患者应用粒细胞集落刺激因子/粒-巨噬细胞集落刺激因子动员后外周血CD34+细胞明显增加[(0.054±0.032)%,(1.82±0.76)%,P＜0.01].22例联合应用粒细胞集落刺激因子和粒-巨噬细胞集落刺激因子动员患者CD34+细胞和粒-巨噬细胞集落形成单位分别为(8.76±3.39)×106/kg,(3.52±1.33)×105/kg,明显高于单用粒细胞集落刺激因子的20例患者[(6.12±2.11)×106/kg,(2.03±1.07)×105/kg,P＜0.05].单独应用粒细胞集落刺激因子及粒细胞集落刺激因子与粒-巨噬细胞集落刺激因子合用后随CD34+细胞增加,T淋巴细胞亚群变化不明显(P＞0.05).②外周血干细胞动员相关不良反应:全部病例出现Ⅱ～Ⅲ度脱发,血小板均有不同程度的下降,为(54.43±26.14)×109L-1,21例患者出现感染性发热(37.8～41.0 ℃),经抗生素治疗感染均在短期内得到控制.13例患者在白细胞快速上升时出现骨骼疼痛(腰骶部为主).③自体外周血干细胞回输量:自体外周血干细胞非程控冷冻-80 ℃保存2.0～6.5个月,细胞回收率(88.7±7.4)%,台盼蓝拒染率(92.1±5.5)%,回输的单个核细胞(5.21±2.44)×108/kg,CD34+细胞(6.89±3.55)×106/kg,粒-巨噬细胞集落形成单位(2.58±2.33)×105/g.④循环血量每次10～16 L(终点分血量均在3个TBV上).无严重毒副反应.26例接受自体外周血干细胞移植者造血功能均获得满意重建.结论:米托蒽醌-大剂量阿糖胞苷方案化疗后单用粒细胞集落刺激因子及粒细胞集落刺激因子与粒-巨噬细胞集落刺激因子合用均能安全、有效动员自体外周血干细胞,但以合用更为有效.大容量采集是提高干细胞产率,减少采集次数的重要手段.
揹景:外週血榦細胞移植成功的首要條件是榦細胞的有效動員和採集,選擇高效低毒的動員方案,掌握動員和採集時機與動員效果密切相關.目的:探討米託蒽醌-大劑量阿糖胞苷方案化療後,單用粒細胞集落刺激因子或粒細胞集落刺激因子與粒-巨噬細胞集落刺激因子閤用對噁性血液病和實體瘤患者自體外週血榦細胞的動員效果.設計:觀察對比實驗.單位:徐州醫學院附屬醫院血液科.對象:選擇1998-09/2006-12在徐州醫學院附屬醫院血液科收治的42例噁性血液病和實體瘤患者,診斷符閤國際白血病分型及世界衛生組織新分類標準.男25例,女17例,年齡7～54歲,平均29歲,體質量(52±18)kg.其中急性髓細胞白血病12例,急性淋巴細胞白血病6例,慢性粒細胞白血病慢性期1例,非霍奇金淋巴瘤15例,霍奇金淋巴瘤4例,多髮性骨髓瘤2例,晚期乳癌2例.患者均經常規化療達到或接近完全緩解,骨髓細胞學檢查無腫瘤細胞浸潤.心、肺、肝、腎等主要髒器功能正常.動員前化療療程平均8次,所有患者均對治療項目知情同意.方法:患者均採用米託蒽醌10 mg/(m2·d)靜脈滴註第2～3 d後,阿糖胞苷2 g/m2靜脈滴註第1～2 d,1次/12 h.噹白細胞計數下降至最低點開始迴升時,20例患者使用粒細胞集落刺激因子5～7.5 μg/(kg·d),連用3～5 d,22例患者早6:00給予粒細胞集落刺激因子5～7.5 μg/(kg·d),晚6:00給予粒-巨噬細胞集落刺激因子5～7 μg/(kg·d).白細胞計數＞2.5×109 L-1,CD34+細胞≥1%時,用CS 3 000 plus血細胞分離機連續2 d採集自體外週血榦細胞,檢測CD34+細胞含量和T淋巴細胞亞群.①單箇覈細胞與FITC標記的CD34+、CD3和CD8單抗及與CD4PE標記的CD4單抗4 ℃混閤30 min,採用流式細胞儀檢測CD34+細胞和T細胞亞群,分析5×105箇細胞,得齣CD3、CD34+細胞含量及CD4/CD8比值.用甲基纖維素法測定粒-巨噬細胞集落形成單位.②觀察術後相關不良反應.③針對不同類型疾病給予相應預處理36～48 h後迴輸自體外週血榦細胞,併行單箇覈細胞計數及檯盼藍染色,解凍後檢測粒-巨噬細胞集落形成單位和CD34+細胞.主要觀察指標:①動員前後CD34+細胞和T細胞亞群變化.②術後相關不良反應.③自體外週血榦細胞迴輸量(單箇覈細胞計數、粒-巨噬細胞集落形成單位和CD34+細胞數).結果:納入患者42例,均進入結果分析.①動員前後CD34+細胞和T細胞亞群變化:患者應用粒細胞集落刺激因子/粒-巨噬細胞集落刺激因子動員後外週血CD34+細胞明顯增加[(0.054±0.032)%,(1.82±0.76)%,P＜0.01].22例聯閤應用粒細胞集落刺激因子和粒-巨噬細胞集落刺激因子動員患者CD34+細胞和粒-巨噬細胞集落形成單位分彆為(8.76±3.39)×106/kg,(3.52±1.33)×105/kg,明顯高于單用粒細胞集落刺激因子的20例患者[(6.12±2.11)×106/kg,(2.03±1.07)×105/kg,P＜0.05].單獨應用粒細胞集落刺激因子及粒細胞集落刺激因子與粒-巨噬細胞集落刺激因子閤用後隨CD34+細胞增加,T淋巴細胞亞群變化不明顯(P＞0.05).②外週血榦細胞動員相關不良反應:全部病例齣現Ⅱ～Ⅲ度脫髮,血小闆均有不同程度的下降,為(54.43±26.14)×109L-1,21例患者齣現感染性髮熱(37.8～41.0 ℃),經抗生素治療感染均在短期內得到控製.13例患者在白細胞快速上升時齣現骨骼疼痛(腰骶部為主).③自體外週血榦細胞迴輸量:自體外週血榦細胞非程控冷凍-80 ℃保存2.0～6.5箇月,細胞迴收率(88.7±7.4)%,檯盼藍拒染率(92.1±5.5)%,迴輸的單箇覈細胞(5.21±2.44)×108/kg,CD34+細胞(6.89±3.55)×106/kg,粒-巨噬細胞集落形成單位(2.58±2.33)×105/g.④循環血量每次10～16 L(終點分血量均在3箇TBV上).無嚴重毒副反應.26例接受自體外週血榦細胞移植者造血功能均穫得滿意重建.結論:米託蒽醌-大劑量阿糖胞苷方案化療後單用粒細胞集落刺激因子及粒細胞集落刺激因子與粒-巨噬細胞集落刺激因子閤用均能安全、有效動員自體外週血榦細胞,但以閤用更為有效.大容量採集是提高榦細胞產率,減少採集次數的重要手段.
배경:외주혈간세포이식성공적수요조건시간세포적유효동원화채집,선택고효저독적동원방안,장악동원화채집시궤여동원효과밀절상관.목적:탐토미탁은곤-대제량아당포감방안화료후,단용립세포집락자격인자혹립세포집락자격인자여립-거서세포집락자격인자합용대악성혈액병화실체류환자자체외주혈간세포적동원효과.설계:관찰대비실험.단위:서주의학원부속의원혈액과.대상:선택1998-09/2006-12재서주의학원부속의원혈액과수치적42례악성혈액병화실체류환자,진단부합국제백혈병분형급세계위생조직신분류표준.남25례,녀17례,년령7～54세,평균29세,체질량(52±18)kg.기중급성수세포백혈병12례,급성림파세포백혈병6례,만성립세포백혈병만성기1례,비곽기금림파류15례,곽기금림파류4례,다발성골수류2례,만기유암2례.환자균경상규화료체도혹접근완전완해,골수세포학검사무종류세포침윤.심、폐、간、신등주요장기공능정상.동원전화료료정평균8차,소유환자균대치료항목지정동의.방법:환자균채용미탁은곤10 mg/(m2·d)정맥적주제2～3 d후,아당포감2 g/m2정맥적주제1～2 d,1차/12 h.당백세포계수하강지최저점개시회승시,20례환자사용립세포집락자격인자5～7.5 μg/(kg·d),련용3～5 d,22례환자조6:00급여립세포집락자격인자5～7.5 μg/(kg·d),만6:00급여립-거서세포집락자격인자5～7 μg/(kg·d).백세포계수＞2.5×109 L-1,CD34+세포≥1%시,용CS 3 000 plus혈세포분리궤련속2 d채집자체외주혈간세포,검측CD34+세포함량화T림파세포아군.①단개핵세포여FITC표기적CD34+、CD3화CD8단항급여CD4PE표기적CD4단항4 ℃혼합30 min,채용류식세포의검측CD34+세포화T세포아군,분석5×105개세포,득출CD3、CD34+세포함량급CD4/CD8비치.용갑기섬유소법측정립-거서세포집락형성단위.②관찰술후상관불량반응.③침대불동류형질병급여상응예처리36～48 h후회수자체외주혈간세포,병행단개핵세포계수급태반람염색,해동후검측립-거서세포집락형성단위화CD34+세포.주요관찰지표:①동원전후CD34+세포화T세포아군변화.②술후상관불량반응.③자체외주혈간세포회수량(단개핵세포계수、립-거서세포집락형성단위화CD34+세포수).결과:납입환자42례,균진입결과분석.①동원전후CD34+세포화T세포아군변화:환자응용립세포집락자격인자/립-거서세포집락자격인자동원후외주혈CD34+세포명현증가[(0.054±0.032)%,(1.82±0.76)%,P＜0.01].22례연합응용립세포집락자격인자화립-거서세포집락자격인자동원환자CD34+세포화립-거서세포집락형성단위분별위(8.76±3.39)×106/kg,(3.52±1.33)×105/kg,명현고우단용립세포집락자격인자적20례환자[(6.12±2.11)×106/kg,(2.03±1.07)×105/kg,P＜0.05].단독응용립세포집락자격인자급립세포집락자격인자여립-거서세포집락자격인자합용후수CD34+세포증가,T림파세포아군변화불명현(P＞0.05).②외주혈간세포동원상관불량반응:전부병례출현Ⅱ～Ⅲ도탈발,혈소판균유불동정도적하강,위(54.43±26.14)×109L-1,21례환자출현감염성발열(37.8～41.0 ℃),경항생소치료감염균재단기내득도공제.13례환자재백세포쾌속상승시출현골격동통(요저부위주).③자체외주혈간세포회수량:자체외주혈간세포비정공냉동-80 ℃보존2.0～6.5개월,세포회수솔(88.7±7.4)%,태반람거염솔(92.1±5.5)%,회수적단개핵세포(5.21±2.44)×108/kg,CD34+세포(6.89±3.55)×106/kg,립-거서세포집락형성단위(2.58±2.33)×105/g.④순배혈량매차10～16 L(종점분혈량균재3개TBV상).무엄중독부반응.26례접수자체외주혈간세포이식자조혈공능균획득만의중건.결론:미탁은곤-대제량아당포감방안화료후단용립세포집락자격인자급립세포집락자격인자여립-거서세포집락자격인자합용균능안전、유효동원자체외주혈간세포,단이합용경위유효.대용량채집시제고간세포산솔,감소채집차수적중요수단.
BACKGROUND: The primary qualification of peripheral blood stem cell transplantation (PBSCT) is the effective mobilization and harvesting of hematopoietic stem cells. The mobilization efficacy is closely related to the selection of high-efficacy low-toxicity regimen, the timing of mobilization and harvesting as well.OBJECTIVE: To investigate the efficacy of mitoxantone (MIT) combined with high-dose arabinosylcytosin (Ara-C),followed by granulocyte colony-stimulating factor (G-CSF) alone or combination of G-CSF and granulocyte-macrophage colony-stimulating factor (GM-CSF) on mobilizing PBSCs in patients with hematological malignancies and solid tumors.DESIGN: Controlled study with observation.SETTTNG: Department of Hematology, the Affiliated Hospital of Xuzhou Medical College.PARTICIPANTS: Forty-two patients with hematological malignancies and solid tumors admitted to Department of Hematology, Xuzhou Medical College from September 1998 to December 2006 were involved in this study. They were diagnosed according to FAB classification criteria and new WHO proposals. The involved patients, 25 male and 17 female, averaged 29 years ranging from 7 to 54 years and weighted (52±18) kg. Among them, 12 were patients with acute myeloblastic leukemia, 6 were patients with acute lymphoblastic leukemia (ALL), 1 was patient with chronic granulocytic leukemia (CGL) at chronic phase, 15 were patients with non-Hodgkin lymphoma (NHL), 4 were patients with Hodgkin lymphoma (HL), 2 was patient with multiple myeloma (MM), 2 were patients with advanced breast cancer. All the patients apprcached to or got complete remission after conventional chemotherapy. No tumor cell infiltration was observed in bone marrow cytological examination. The functions of the main organs such as heart, lung, liver and kidney,and so on, were normal. The patients underwent an average of 8-course chemotherapy before the mobilization. Informed consents of all the patients were obtained.METHODS: MIT was intravenously injected at 10 mg/(m2·d)for 2 to 3 days, then Ara-C was also intravenously injected at 2 g/m2 every 12 hours for 1 to 2 days. When white blood cell (WBC) count recovered from the lowest value, 5 to 7.5 μg/ (kg·d)G-CSF was applied in 20 patients for 3 to 5 days successively. And 5 to 7.5 μg/ (kg·d)G-CSF and 5 to 7 μ g/(kg·d)GM-CSF were applied in another 22 patients at 6:00 in the morning and in the evening, respectively. PBSCs harvesting started when WBC ＞ 2.5×109 L-1, especially when CD34+ cells≥ 1%,WBC was doubly increased. Autologous peripheral blood mononuclear cells (MNCs) were collected with CS3000 plus blood cell separator for detecting the level of CD34+ cells and T lymphocyte subsets. CFU-GM assays were performed in a methyl-cellulose-based clonogenic assay.① MNCs mixed with FITC-labeled CD34+, CD3 and CD8 monoclonal antibodies as well as CD4 PE-labeled CD monoclonal antibody at 4 ℃ for 30 minutes. 5×105 cells were determined, and CD3 and CD34+ levels, CD4/CD8 were determined by flow cytometer.Colony forming unit-granulocyte macrophage (CFU-GM) was determined with methyl cellulose. ② Related adverse reactions were observed after operation. ③ Aiming to different types of diseases,autologous PBSCs were back infused 36 to 48 hours after pre-disposal treatment. MNCs count and trypan-blue drying were done. Levels of CFU-GM and CD34+ cells were determined after unfreezing.MATN OUTCOME MEASURES: ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization. ② Postoperative related adverse reactions. ③ Back perfusion volume of autologous PBSCs (MNCs count, the number of CFU-GM and CD34+ cells).RESULTS: Forty-two involved patients participated in the final analysis. ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization: Without using hematopoietic growth factors (HGF), the percentage of CD34+ cells in peripheral blood of the patients was (0.054±0.032)%. After using G-CSF/GM-CSF treatment, it was (1.82±0.76)%,which was obviously increased compared with that of without using HGF (P ＜ 0.001). The CD34+ cells and CFU-GM yields of 22 patients in C-CSF plus GM-CSF combination group [(8.76±3.39)×106/kg, (3.52±1.33)×105/kg, respectively]were significantly higher than those of 20 patients in G-CSF alone group [(6.12±2.11)×106/kg, (2.03±1.07)×105/kg,respectively (P ＜ 0.05)]. There were no obvious changes of T lymphocyte subsets in the patients when using G-CSF/GM-CSF for some days except that CD34+ cells increased gradually (P ＞ 0.05). ② Postoperative related adverse reactions: Ⅱ to Ⅲ degree hair-loss was seen in all the patients. Blood platelets dropped to (54.43±26.14)×109 L-1 at different degrees. Infective fevers (37.8 ℃ to 41.0 ℃) occurred in 21 patients. But they were controlled in short term after antibiotics treatment. All the side effects of G-CSF and GM-CSF were mild and reversible, easily controlled with paracetamol or steroids. Bone pain (mainly in lumbosacral region) occurred in 13 patients when WBC went up quickly. ③ Back perfusion volume of autologous PBSCs: PBSCs were cryopreserved at -80 ℃ without program control for 2.0 to 6.5 months. The cell recovery rate was (88.7±7.4) %. Trypan blue exclusion rate was (92.1±5.5) %. The back perfusion volume of MNCs, CD34+ cells and CFU-GM yields were (5.21±2.44)×108/kg, (6.89±3.55)×106/kg, (2.58±2.33)×105/kg,respectively. ④Circulation blood volume were 10 to 16 L (end-point separation blood volume were all above trebling TBV). Hematopoiesis was well reconstituted in 40 patients received autologous PBSCT.CONCLUSTON: MIT and high-dose Ara-C chemotherapy combined with both G-CSF alone and G-CSF plus GM-CSF can safely and effectively mobilize autologous PBSCs, while G-CSF plus GM-CSF is superior to G-CSF alone.Large-volume leukapheresis is an important method to enhance the productive rate of stem cells and decrease the times of harvesting.
