白细胞介素11联合粒细胞集落刺激因子对小鼠异基因骨髓移植后移植物抗宿主病影响的研究  

Significance of donors treatment with combination of recombinant human interleukin-11 and recombinant human granulocyte-colony stimulating factor in reduction of incidence of lethal grafts versus host disease after allogeneic bone marrow transplantation

作　　者：赵杰[1] 赵翔宇[1] 黄晓军[1] 

机构地区：[1]北京大学人民医院血液病研究所,100044

出　　处：《中华医学杂志》2005年第35期2497-2502,共6页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30370591);卫生部科研基金资助项目(200363);教育部"211"工程基金资助项目;教育部高等学校博士学科点专项科研基金资助项目(20020001086);新世纪优秀人才支持计划基金资助项目(NCET-04-0011)

摘　　要：目的探讨重组白细胞介素11(rhIL-11)联合重组粒细胞集落刺激因子(rhG-CSF)用于供者对小鼠异基因骨髓移植后移植物抗宿主病(GVHD)的作用及其机制。方法建立同种异基因骨髓移植急性GVHD模型,以C57BL/6H-2b为供鼠,rhIL-11、rhG-CSF或二者联合应用作为动员剂,BALB/CH-2d受鼠经致死剂量(8·0Gy)照射后随机分为5组,A组为单纯照射组(10只),B组为GVHD对照组(18只),C组为G-CSF移植组(18只),D组为IL-11移植组(18只),E组为联合移植组(18只),在照射后4h移植供鼠骨髓细胞和脾细胞。观察移植后小鼠的生存期和病死率,应用流式细胞仪测定嵌合体形成情况,用四甲基偶氮唑盐(MTT)法检测混合淋巴细胞反应,酶联免疫吸附实验(ELISA)检测白细胞介素4(IL-4)、γ干扰素(IFN-γ)等细胞因子的分泌情况,并进行组间比较。结果E组小鼠生存时间明显长于其他各组(B、C、D),差异有统计学意义(P<0·01),30d存活率B、C、D、E组分别是:0、27·3%(3/11)、36·4%(4/11)、63·6%(7/11)。B组小鼠均出现GVHD病理改变,其程度与发生GVHD的时间密切相关,GVHD发生越早,病理改变越重;E组有3只小鼠在移植后20d内发生GVHD,病理程度改变较其他各组轻。与B、C、D组相比,E组移植后14d脾细胞培养上清中IFN-γ水平较低(92pg/ml±14pg/mlvs172pg/ml±31pg/ml、132pg/ml±23pg/ml、117pg/ml±28pg/ml,均P<0·01),IL-4的水平较高(36pg/ml±7pg/mlvs16pg/ml±4pg/ml、26pg/ml±4pg/ml、24pg/ml±4pg/ml,均P<0·05),D、E组肿瘤坏死因子α(TNF-α)水平相近,但明显低于B、C组(均P<0·01),并且E组对宿主抗原的淋巴细胞增殖均低于B、C、D组(0·22±0·08vs0·87±0·14、0·54±0·21、0·49±0·18,均P<0·01)。结论rhIL-11联合rhG-CSF用于供者能产生协同作用,减轻GVHD,其机制可能与受者体内T细胞向Th2细胞因子极化、炎症介质如TNF-α降低有关。Objective To investigate the effects of donor treatment with combination of recombinant human interleukin-11 （rhIL-11 ） and recombinant human granulocyte-colony stimulating factor （rhG-CSF） on incidence of lethal graft versus host disease （GVHD） after allogeneie bone marrow transplantation （BMT）. Methods Forty C57BL/6 mice, as donors, were treated with phosphate buffered saline （PBS） , or optimal doses of rhG-CSF, rhIL-11, or rhG-CSF ＋ rhIL-11 for five consecutive days. Fifty BALB/C mice, as recipients, received a lethal dose of 8. 0 Gy total body irradiation （TBI） 4 -6 h before the transplantation. Ten C57BL/6 mice without transplantation were used as pure radiation control group （group A）. The other 40 C57BL/6 mice received the allogeneic bone marrow from the donors treated with PBS （group B）, rhG-CSF （group C）, rhIL-11 （group D）, or rhG-CSF ＋ rgIL-11 （group E） 14 and 30 days after the transplantation, suspensions of bone marrow cells wgroupere prepared from the 4 groups. Monoclone antibody of PE-H-2D^b was added. The percentage of the H-2D^b positive cells was observed by flow cytometry to detect the chimerism to determine the proportion of the cells of donor origin. The liver, spleen, intestine, and skin tissues of the mice presenting symptoms of GVHD were taken out before the diseased mice died. The long-surviving mice were killed 60 days after the transplantation and the above-mentioned tissues were taken out. Light microscopy was used to observe the pathological changes. 14 days after transplantation, the spleen cells of the different groups were obtained to undergo mixed leucocyte reaction （ MLR）, and 14 days after transplantation another spleen cells of groups B, C, D, and E were obtained to prepare suspension of mononuclear cells to test the levels of IL-4, interferon （ IFN）-γ and tumor necrosis factor （ TNF）-α by ELISA. Result The survival time of group E was 38 d ± 19 d, significantly longer than the other groups （all P 〈0.01 ）. The

关 键 词：白细胞介素1 1 移植物抗宿主病 粒细胞集落刺激因子 骨髓移植 重组粒细胞集落刺激因子 同种异基因骨髓移植 白细胞介素11 联合移植 移植后 

分 类 号：R392[医药卫生—免疫学]