应用含标记基因K562细胞制备的小鼠白血病模型

背景:文献报道应用NOD/SCID及SCID小鼠可以建立移植性人白血病小鼠模型,但由于小鼠存在免疫缺陷,使得在自体干细胞移植及移植后相关过继免疫治疗方面的研究受到限制和影响。目的:探索用SPF级Balb/c小鼠和转染GFP及NeoR基因的K562细胞株制备白血病模型的方法。方法:实验分为5组,A、B组和C、D组分别经X射线照射2Gy和3Gy,24h后取对数生长期的K562(GFP+/Neo+)细胞,A、C组尾静脉注射2×106个/只;B、D组尾静脉注射5×106个/只;E组为正常对照组。观察小鼠生存时间,进行骨髓细胞及外周血白细胞分类,采用流式细胞仪测定GFP阳性细胞及PCR方法测定Neo基因。结果与结论:实验各组在接种5~7d时发病,分别于30,23,24,17d内全部死亡;生存天均显著短于正常对照组(P0.01)。体质量均较正常对照组显著下降(P0.05)。小鼠的白血病发病率为100%,无自发缓解。随接种细胞数量的增加和照射剂量的增加,小鼠的存活时间缩短,且外周血及骨髓中白血病细胞所占比例增加。流式细胞仪测定及PCR方法也证实了GFP+细胞和NeoR基因在肝、脾中的存在。结果证实Balb/c小鼠经照射后从尾静脉注射K562细胞株可以制备获得白血病小鼠模型。     

自体造血干细胞移植联合异体细胞因子诱导的杀伤细胞及白细胞介素2治疗高危急性髓系白血病2例*

背景：自体造血干细胞移植治疗高危急性髓系白血病的复发率极高,如何降低移植后复发率至今仍是难点。 目的：观察自体造血干细胞移植联合异体细胞因子活化杀伤(CIK)细胞及白细胞介素2治疗高危急性髓系白血病的临床效果。 方法：2例高危急性髓系白血病患者,经诱导化疗及巩固强化治疗后,在第1次缓解期行自体造血干细胞移植, 移植后1个月给予三四疗程异体CIK细胞输注,每隔半年1个疗程,每疗程分5次输注异体CIK细胞,每隔1日输注1次。每次输注CIK细胞前半小时内给予皮下注射白细胞介素2,第1次输注后隔日皮下注射白细胞介素2,半年后减为隔2日1次,1年后减为隔3日1次,1年半后减为1次/周,维持半年后结束,预防白血病复发。 结果与结论：2例患者自体造血干细胞移植后输注异体CIK细胞及皮下注射白细胞介素2无发热、寒战、皮疹等不良反应,无骨髓抑制及移植物抗宿主反应,治疗安全。2例患者持续缓解时间分别为20个月及2年,目前仍无复发。首次得出对于无合适供者的高危急性髓系白血病患者,在缓解后可行自体造血干细胞移植联合异体CIK细胞及白细胞介素2治疗,有机会获得长期无病生存。       

同基因造血干细胞移植免疫重建诱导小鼠皮肤移植耐受

背景：器官移植耐受的最佳效果是能够诱导对移植抗原的特异性免疫耐受。 目的：探讨小鼠异基因皮肤移植后,通过受体同基因造血干细胞移植重建免疫系统诱导移植皮肤免疫耐受的可行性。 方法：取BALB/c小鼠骨髓。以C57BL/6小鼠为供体,BALB/c小鼠为受体,进行异基因皮肤移植;32只受体鼠随机均分为4组：移植对照组、环孢素A组、照射组和骨髓移植组。 结果与结论：照射组小鼠10 d内全部死亡,外周血白细胞数呈持续性降低;而骨髓移植组小鼠长期存活,白细胞数全身照射后6 d降到最低,之后持续性增高,照射后21 d与环孢素A组比较差异无显著性意义(P > 0.05),移植皮肤存活时间显著长于其他各组(P < 0.01),其淋巴细胞浸润及组织结构破坏明显减少,小鼠脾细胞对供体小鼠脾细胞增殖反应显著降低。说明同基因骨髓细胞移植重建免疫系统可显著延长小鼠移植皮肤存活时间,可诱导供者特异性免疫耐受。     

髓腔内骨髓移植诱导异基因小鼠皮肤移植耐受的研究

探讨髓腔内骨髓移植(IBM-BMT)对异基因小鼠皮肤移植耐受的诱导效果。受鼠为雌性C57BL/6(H-2b,B6)小鼠,于第0天接受60Coγ射线全身照射(TBI),4 h内输注雄性BALB/c(H-2d)小鼠来源的骨髓细胞(BMC),2 d后腹腔注射环磷酰胺(CTX)。通过皮肤移植、混合淋巴细胞反应(MLR)检测耐受状态,并通过骨髓染色体分析了解BMC的植入程度。结果显示,接受IBM-BMT的B6小鼠对BALB/c小鼠的皮肤移植物平均存活时间(MST)超过300 d,显著长于其余两组(P<0.001);MLR结果证明,实验组B6小鼠获得供体特异性耐受,在耐受小鼠骨髓中可检测到一定比例的Y染色体存在。以上表明髓腔内骨髓移植可以保证异基因骨髓细胞的植入并形成混合嵌合状态,从而有效地诱导免疫耐受。     

MHC相合造血干细胞移植后cGVHD小鼠模型建立

目的建立主要组织相容性复合物（MHC）相合造血干细胞移植的慢性移植物抗宿主病（Chronic graft versehost disease,cGVHD）小鼠模型,为进一步研究cGVHD的发病机制及防治方法构建平台。方法以雄性DBA/2H-2d为供鼠,雌性BABL/CH-2d为受鼠,接受^60Co5.5Gy全身照射后分成3组,4～6h内完成移植。A组输注RPMI1640培养液、B组输注5×10^7个脾细胞/只、C组输注10×10^7个脾细胞/只。观察指标为：①造血重建时间;②移植物植入情况;③各组小鼠cGVHD的临床表现;④各组的病理学改变及成模率。结果①造血重建：C组较A、B组造血重建延迟,差异有统计学意义。②植入情况检测：移植组小鼠均为供、受者共存的混合嵌合体。③cGVHD的临床表现及评分：A组均未出现cGVHD的临床表现,B、C组小鼠出现cGVHD的临床表现,两组的临床评分有统计学差异（P=0.016）。④病理改变及成模率：A组病理学未见明显异常;B、C两组出现cGVHD的病理学改变,但是两组评分无统计学差异（P=0.410）;根据病理学评分判断各组小鼠的成模率分别为0%、60%、80%。结论 60Co5.5Gy全身照射后输注5×10^7个脾细胞可诱导出cGVHD小鼠模型,且配型方式与临床MHC相合造血干细胞移植类似,可以作为研究MHC相合造血干细胞移植后cGVHD的理想模型。     

雌性小鼠骨髓移植给雄性小鼠后内源性骨髓细胞残存状态的研究

 目的:以雌性小鼠骨髓移植给雄性小鼠的方法,通过检测雄性小鼠血细胞的Y染色体来明确内源性骨髓细胞的残存状态。方法:将雌性或雄性C57BL/6小鼠作为受体,实验组用［137Cs］照射,6 h后每只经尾静脉注射供体小鼠骨髓细胞1×107。统计骨髓移植后14 d动物的存活率,并通过眶静脉采血观察外周血白细胞数量的变化,检测受体雄性小鼠体内Y染色体基因水平的变化以明确骨髓移植的效果。结果:分别用1 000、950和900 rad的照射剂量对受体小鼠进行照射后将供体小鼠的骨髓移植到受体小鼠体内,1 000和950 rad剂量时雌性受体小鼠可迅速恢复造血功能,而雄性受体小鼠则仅有48%的存活率。900 rad照射剂量骨髓移植后,雄性受体小鼠迅速恢复了造血功能,13 d后外周血白细胞计数基本恢复正常。移植后的雄性受体小鼠在5周内已检测不到外周血细胞Y染色体基因,表明雄性受体小鼠的骨髓被完全破坏,雌性供体小鼠的骨髓可完全替代受体雄性小鼠的骨髓并且发挥造血功能。结论: 在照射剂量900 rad照射后,雄性小鼠可以作为骨髓移植受体,为将来应用雄性小鼠作为骨髓移植受体动物开展有关心血管疾病的研究奠定了实验基础。     

同基因造血干细胞移植延长同种异体小鼠皮肤移植物存活时间的实验研究

为了研究同基因造血干细胞移植诱导器官移植免疫耐受的可行性。建立小鼠异基因皮肤移植模型,术后2周给予FK506腹腔注射,3周起行全身照射及同基因骨髓移植,观察记录小鼠和移植物存活情况,以流式细胞检测受体GFP嵌合表达,混合淋巴细胞反应、迟发型超敏反应检测诱导耐受的特异性和效能。实验组小鼠移植物存活时间达(29.14±4.92)d,显著长于对照组(P<0.05);GFP在BMT后4周、6周嵌合程度达到82%、91%;实验组MLR、DTH结果与对照组差异显著,提示诱导耐受具有高度特异性和高效性。同基因造血干细胞移植联合免疫抑制剂治疗可以有效诱导小鼠皮肤移植的免疫耐受。     

NK细胞在异基因骨髓移植中作用的动物实验研究

本文探讨NK细胞在异基因小鼠骨髓移植GVHD和GVL反应中的作用。受体小鼠以60 Co照射 ,总剂量 8 0Gy;骨髓细胞经尾静脉注入受体小鼠中 ;GVHD反应指标包括 (1)弓背体位、大小便及脱毛等一般表现 ;(2 )嵌合体、体内混合淋巴细胞反应、 30d生存率为检测GVHD反应的定量指标 ;GVL作用指标包括外周血白细胞总数及白血病细胞比例 ,脾脏白血病细胞浸润比例 ,以及死亡率观察 ;病理学检查作为鉴别GVL实验中是否主要死于GVHD的指标。GVHD反应结果表明 ,异基因与后输NK细胞的异基因骨髓移植组均于 15d内死亡。而先输NK细胞组的GVHD反应明显减轻 ,30d死亡率为5 1%。同时输NK细胞组 30d内死于GVHD反应。移植后第 7天与 15d时 ,以先输NK细胞组的嵌合率高 ,第 30天时各组均未检出明显嵌合体存在。GVL作用实验方面 ,异基因、同时输NK细胞组及后输NK细胞的骨髓移植组虽然于 12d内表现出抗白血病作用 ,但在 30d内相继死于GVHD反应。先输NK细胞组于移植后 12d开始观察到较明显GVHD样反应 ,而外周血或脾细胞中L615细胞比例变化结果证实该移植途径具有较好的抗白血病作用。先输注供体NK细胞的异基因骨髓移植方式既能减轻GVHD反应 ,又能保留较好的GVL作用     

MHC半相合骨髓移植后慢性GVHD小鼠模型的建立

目的建立MHC半相合异基因骨髓移植(allo-BMT)后慢性GVHD小鼠模型。方法以(Balb/c×C57BL/6)F1H-2d/b(CB6F1)雌性小鼠为受者,预处理条件为不同剂量的全身照射(TBI,60Co照射),输注MHC半相合Balb/cH-2d雄性小鼠骨髓细胞与不同数量脾细胞,观察移植后小鼠体质量变化,靶器官病理变化,血清抗ssDNA抗体、抗dsDNA抗体。结果在照射剂量为8Gy,输注骨髓细胞数量为8×106、脾细胞数量为4.5×107的小鼠至实验结束(移植后100d)全部存活,体质量减轻与对照组和其它实验组相比差异显著(P<0.05)。血清抗ssDNA抗体、抗dsDNA抗体较对照组和其它实验组显著升高(P<0.05)。该组皮肤、肝脏等病理改变明显。结论在照射剂量为8Gy,输注骨髓细胞数量为8×106、脾细胞数量为4.5×107的小鼠成功诱导出半相合allo-BMT慢性GVHD,为进一步研究慢性GVHD的发病机理、生物学特性、干预因素等打下了重要基础。     

自体造血干细胞移植治疗恶性淋巴瘤后大剂量白细胞介素2过继免疫对长期生存的影响

背景：过继免疫治疗是目前肿瘤免疫治疗的热点,白细胞介素2是一种具有多种生物学活性的细胞因子,在机体的抗肿瘤免疫中起到重要作用。 目的：评价比较淋巴瘤自体造血干细胞移植治疗后应用与不应用大剂量白介素2行免疫治疗的临床疗效。 方法：回顾分析30例恶性淋巴瘤患者(治疗组)自体造血干细胞移植后行大剂量白细胞介素2 治疗,与随机挑选30例患者(对照组)自体造血干细胞移植后未行白细胞介素2治疗进行对比,检测两组患者外周血T淋巴细胞亚群,观察两组免疫功能的变化,并对所有患者进行随访观察。 结果与结论：自体造血干细胞移植后白细胞介素2治疗组外周血T淋巴细胞亚群CD3⁺、CD4⁺、CD8⁺、CD4⁺/CD8⁺水平明显提升。随访结束时统计复发率：治疗组13.3%,对照组26.7%;中位生存期：治疗组14~98 (42±2)个月,对照组8~78 (28±2)个月。提示恶性淋巴瘤自体造血干细胞移植后行大剂量白细胞介素2治疗能提高患者的免疫功能,减少移植后复发率,并有望延长生存期。     

Effects of intrabone marrow-bone marrow transplantation plus adult thymus transplantation on survival of mice bearing leukemia

We recently found that allogeneic intrabone marrow-bone marrow transplantation (IBM-BMT) plus adult thymus transplantation (ATT) from the same donor is effective in mice bearing solid tumors. In the current study, we examined the effects of this strategy on the survival of mice with leukemia. One week after intravenous injection of 1×10(6) leukemic cells (EL-4, H-2(b)) into 8-week-old B6 (H-2(b)) mice, the mice were 8 Gy irradiated and transplanted with 1×10(7) bone marrow cells (BMCs) from 8-week-old BALB/c mice (H-2(d)) by IBM-BMT with or without donor lymphocyte infusion (DLI) or ATT. All the mice without treatment died within 70 days after injection of EL-4. About 40% of those treated with IBM-BMT alone died within 100 days due to tumor relapse. In contrast, those treated with IBM-BMT+DLI or ATT showed the longest survival rate without relapse of leukemia. In addition, the former showed less graft versus host disease (GVHD) than the latter. The mice treated with IBM-BMT+ATT also showed an intermediate percentage of effector memory (EM) and central memory (CM) cells between those treated with BMT alone and those treated with IBM-BMT+DLI. The numbers and functions of T cells increased in those treated with IBM-BMT+ATT with interleukin-2 and interferon-γ production. These results suggest that IBM-BMT+ATT is effective in the treatment of leukemia with strong graft versus leukemia without increased risk of GVHD.     

Effect of NK cells on GVHD in H-2 haploidentical bone marrow transplantation in mice

Objective: To study the effect of natural killer (NK) cells on graft-versus-host disease (GVHD) after H-2 haploidentical bone marrow transplantation (BMT) in mice. Methods :Murine model of H-2 haploidentical BMT was established by using Balb/c (H-2d) mouse as recipient, and Balb/c(H-2d)×C57BL/6 (H-2b) (H-2d/b) mouse as donor. Lethally irradiated Balb/c (H-2d) mice were transplanted with the bone marrow cells from Balb/c(H-2d)×C57BL/6(H-2b) (H-2d/b) mice containing donor spleen cells and/or NK cells. GVHD and survival rates were studied by observation of clinical manifestations and pathological changes. Results:In the group of bone marrow +spleen cells, GVHD was induced in 90% mice; but in the group plus with low amount of NK cells,GVHD was induced in 20% mice; and in the group transplanted with high amount of NK cells, GVHD was induced only in 10% mice. Compared to the group transplanted only with BM plus spleen cells, the incidences of GVHD in the latter two groups decreased significantly (P＜0.01) and the survival rates at different periods of 15, 30, 45 and 60 days increased obviously (P＜0.01 ). Conclusion: In mouse H-2 haploidentical BMT, alloreactive NK cells can reduce the incidence of GVHD and increase the survival rate.     

在小鼠异基因骨髓细胞移植中超抗原SEB诱导的耐受特征

目的:研究在异基因骨髓细胞移植中葡萄球菌肠毒素B(SEB)诱导的耐受强度和细胞学特征。方法:选用C57BL/J小鼠作为受体,BALB/c小鼠作为供体。所有受体小鼠在接受6×107骨髓细胞前均接受6.0Gy60Coγ射线的照射,随机分成3组:组1为只接受6.0Gy60Coγ射线照射的对照组(照射对照组,RI);组2为照射后注射生理盐水(移植对照组,Tran.);组3为照射后注射60μgSEB(SEB组)。180d后由组2和组3实验鼠获得两组C57BL/L-BALB/c嵌合体小鼠。用流式细胞术分析移植后30~180d受体小鼠体内CD4 T、CD8 T、CD3 /NK1.1 NKT淋巴细胞亚群的数量和MHCH-2Kb、H-2Kd抗原表达的百分率,用MLR方法测定嵌合体小鼠淋巴细胞对ConA和异源性抗原的反应性。结果:(1)接受大剂量骨髓细胞移植后,注射SEB和注射生理盐水的两组小鼠均可存活180d以上,SEB组小鼠呈现出BALB/c供体小鼠的颜色特征(白色);Tran.组小鼠呈现出其灰白色。(2)SEB组嵌合体小鼠对ConA的反应性明显低于RI组和Tran.组;对异源性抗原的应答高于Tran.组。(3)SEB组小鼠外周血中CD4 T细胞的数量在移植后30~60d明显下降,随后增加;CD8 T细胞的数量不变。CD3 /NK1.1 NKT细胞的数量,从接受移植后30d开始增加,随着时间的延长而递增,到180d达到5.71%。存活180d的嵌合体小鼠,体内供体小鼠特有的MHCH-2Kd抗原的表达率高达80.95%,自体MHCH-2Kb抗原表达的百分率只占1.45%。(4)与SEB组相反,Tran.组小鼠CD8 T细胞的数量持续下降;CD3 /NK1.1 NKT细胞的数量的增加只在移植后180d上升达到5.07%。结论:在异基因骨髓细胞移植中,SEB诱导的耐受性比单纯骨髓移植诱导的耐受性强。SEB诱导的耐受性与移植早期特异性CD4 T细胞数量的减少和NKT细胞数的持续增加有关。反应性的降低表现为T细胞对ConA反应性的下降。     

Enhancement of allogeneic hematopoietic stem cell engraftment and prevention of GVHD by intra-bone marrow bone marrow transplantation plus donor lymphocyte infusion

We examined the effect of intra-bone marrow (IBM)-bone marrow transplantation (BMT) in conjunction with donor lymphocyte infusion (DLI) on the engraftment of allogeneic bone marrow cells (BMCs) in mice. Recipients that had received 6 Gy of radiation completely rejected donor BMCs, even when IBM-BMT was carried out. However, when BMCs were IBM injected and donor peripheral blood mononuclear cells (PBMNCs) were simultaneously injected intravenously (DLI), donor cell engraftment was observed 7 days after BMT and complete donor chimerism continued thereafter. It is of interest that the cells of recipient origin did not recover, and that the hematolymphoid cells, including progenitor cells (Lin-/c-kit+ cells) in the recipients, were fully reconstituted with cells of donor origin. The cells in the PBMNCs responsible for the donor BMC engraftment were CD8+. Recipients that had received 6 Gy of radiation, IBM-BMT, and DLI showed only a slight loss of body weight, due to radiation side effects, and had no macroscopic or microscopic symptoms of graft-versus-host disease. These findings suggest that IBM-BMT in conjunction with DLI will be a valuable strategy for allogeneic BMT in humans.     

NK细胞在小鼠异基因骨髓移植中的作用

目的:研究自然杀伤(NK)细胞在异基因骨髓移植中对移植物抗宿主病(GVHD)、移植排斥、骨髓植入及造血重建的影响。方法:以近交系小鼠C57/6j(H-2^b)为供鼠、BALB/c(H-2^d)为受鼠,在移植物中增加供者的外周T细胞和/或NK细胞进行异基因骨髓移植,用流式细胞仪检测受鼠的CD₃₄^＋细胞计数和H-2K^b＋细胞表达水平,血细胞自动分析仪检测外周血白细胞计数,并结合临床表现和病理检查,比较不同移植组的存活率、GVHD、植入水平及造血重建等。结果:增加NK细胞组的小鼠存活率显著大于不增加NK细胞组,小鼠出现GVHD的数量少、程度轻,外周血白细胞及骨髓CD₃₄^＋细胞恢复快、H-2K^b＋细胞表达水平高。结论:NK细胞抑制小鼠异基因骨髓移植中的GVHD和移植排斥,促进骨髓植入及造血重建。     

Antileukemic effect of interleukin-2-transduced murine bone marrow after autologous transplantation.

Myeloablative chemotherapy or radiation therapy supported by autologous stem cell transplantation (SCT) for the treatment of hematologic malignancies such as acute leukemia, lymphoma, and myeloma is associated with high rates of relapse. The reasons for this are 1) autologous transplantation lacks the in vivo graft-vs.-tumor (GVT) effect associated with allogeneic SCT, which is effective in controlling or eliminating residual malignant cells remaining in the body after high-dose therapy, and 2) contaminating malignant cells in the autologous graft are reinfused into the body. Some researchers have attempted to administer immunomodulatory cytokines to simulate a GVT effect, and although this has shown some efficacy, it has several disadvantages. These include high toxicity associated with systemic administration, a short in vivo half-life, and insufficient levels reaching the site of residual disease. As an alternative, we investigated whether delivery of the cytokine interleukin (IL)-2 to the bone marrow can exert an antileukemic effect while avoiding the problems associated with systemic administration. We describe the delivery of IL-2 to the bone marrow by transplantation of syngeneic bone marrow, retrovirally transduced with the gene for IL-2, into lethally irradiated mice. We were able to efficiently transduce murine bone marrow with the IL-2 gene without adversely affecting clonogenic output from hematopoietic progenitors, and we were able to achieve expression of the transgene in transplanted animals. However, IL-2 transduction inhibited hematopoietic reconstitution in lethally irradiated mice. Marrow transduced with high-titer, high-expressing IL-2 retrovirus failed to engraft, and a low-titer, low-expressing IL-2 retrovirus also demonstrated reduced engraftment, although engraftment was sufficient to support survival of transplanted mice. Long-term, low-level expression of the IL-2 transgene was detectable in these mice and was effective in exerting an antileukemic effect. Mice transplanted with control marrow and challenged with leukemic cells suffered 100% mortality within 70 days, whereas mice transplanted with IL-2-transduced marrow exhibited 50% survival over the 175-day duration of this study. The work shows that delivery of immunomodulatory cytokines to the bone marrow can be achieved by transplantation of genetically modified hematopoietic cells. Furthermore, low-level IL-2 expression can exert an antileukemic effect. These data suggest that this may be an effective immunotherapeutic strategy to reduce relapse after autologous transplantation, but the selection and expression of the cytokine must be carefully considered to minimize adverse effects on hematopoiesis.     

A new strategy for treatment of malignant tumor: intra-bone marrow-bone marrow transplantation plus CD4- donor lymphocyte infusion

Donor lymphocyte infusion (DLI) is clinically used for the treatment of malignant tumors. We have found recently that intra-bone marrow-bone marrow transplantation (IBM-BMT) can be used to treat various autoimmune diseases, even when radiation doses are reduced. In addition, recently we have found that IBM-BMT can prevent not only graft failure but also graft-versus-host disease (GvHD). Based on these findings, we attempted to prevent and treat the progression of a tumor (Meth-A cell line: BALB/c-derived fibrosarcoma) by DLI plus IBM-BMT. When the tumors had grown to approximately 10 x 10 mm, the tumor-bearing BALB/c (H-2(d)) mice were irradiated with 5 Gy, and whole spleen cells from C57BL/6J (B6) (H-2(b)) mice (as DLI) were then intravenously injected into the BALB/c mice. Simultaneously, bone marrow cells (BMCs) from B6 mice were injected directly into the bone marrow cavity of the BALB/c mice (IBM-BMT). The tumors decreased in size, but the mice died of GvHD. However, when CD4(+) T-cell-depleted spleen cells were used for DLI, the recipients showed only mild GvHD and survived longer, due to the slow growth of the tumor. In contrast, when CD8(+) T-cell-depleted spleen cells were used for DLI, the recipients showed more severe GvHD than those injected with whole spleen cells. These results suggest that IBM-BMT plus DLI (the depletion or reduction of a certain cell population like CD4(+) T cells) could be helpful to suppress both GvHD and tumor growth.     

异基因骨髓移植小鼠免疫功能缺损机制的探讨

异基因骨髓移植(Allogeneic Bone Marrow Transplantation,ABMT)后,受体的免疫功能长期缺损,是患者术后极易感染死亡的重要原因之一.本文对ABMT小鼠(C57BL/6→BALB/c)免疫功能缺损的机制进行了探讨,发现ABMT小鼠IL-2产生明显受损;其脾细胞与(C57BL/6小鼠脾细胞一起过继转移到致死量照射的BALB/C小鼠体内,能抑制移植物抗宿主病(GVHD)的发生.去除其脾T细胞后,这种抑制作用丧失,ABMT小鼠脾细胞上清中发现一种非特异的抑制因子,能抑制正常小鼠脾细胞产生混合淋巴细胞反应的能力;能抑制正常小鼠的脾细胞产生IL-2;抑制正常小鼠脾细胞毒T淋巴细胞(CTL)的杀伤活性.用抗Thy-1.2单抗和补体去除ABMT小鼠脾T细胞后,其脾细胞培养上(?)的上述抑制活性丧失.这说明ABMT小鼠脾T(?)细胞活性增强是其免疫功能缺损的重要原因之一,它通过释放非特异的抑制因子执行其免疫抑制功能.     

Spermine dialdehyde,a novel ex vivo purging agent for both allogeneic and autologous bone marrow transplantations

The use of ex vivo purging agents has shown to be beneficial for both allogeneic and autologous bone marrow transplantations. We have shown previously that spermine dialdehyde (SDA), an oxidized product of spermine, when used in phosphate-buffered saline (PBS) preferentially inhibits T-cell proliferation while sparing myeloid cells. Lethally irradiated mice were rescued by reconstitution with SDA-treated allogeneic marrow and showed no sign of graft-vs-host disease (GVHD). In this paper, we show by HPLC analysis that SDA degraded rapidly in PBS but remained intact in saline. Administered in saline, SDA was more inhibitory on leukemic cell lines than normal myeloid or T-cells. Lethally irradiated mice receiving a syngeneic bone marrow leukemic cell mixture treated ex vivo with SDA in saline did not manifest leukemia. Thus, the preferential inhibitory effect of SDA and its degraded products in different buffers suggest that SDA could act as a novel purging agent for both allogeneic and autologous bone marrow transplantations.     

Combination of intra-bone marrow-bone marrow transplantation and subcutaneous donor splenocyte injection diminishes risk of graft-versus-host disease and enhances survival rate

The combination of allogeneic bone marrow transplantation (allo-BMT) and donor lymphocyte infusion (DLI) is a useful method for establishing donor chimerism and preventing a relapse of leukemia/lymphoma. However, there is a risk of inducing uncontrollable fatal graft-versus-host disease (GVHD). In fact, allo-BMT plus intravenous (IV)-DLI using donor splenocytes induces fatal GVHD in recipient mice. In this study, we examined the effects of the combination of intra-bone marrow (IBM)-BMT and the subcutaneous injection of donor splenocytes (SC-DLI) on the allo-BMT system. Recipient BALB/c mice were conditioned by sublethal irradiation (5 Gy), followed by IBM-BMT plus IV-DLI or SC-DLI in C57BL/6 mice. The IV-DLI group showed better engraftment of donor hemopoietic cells than the control group (without DLI) but showed fatal GVHD. The SC-DLI group, however, showed good reconstitution and mild GVHD. These results suggest that the combination of SC-DLI and IBM-BMT promotes the reconstitution of hemopoiesis and helps reduce the risk of GVHD.