粒细胞集落刺激因子对急性辐射损伤小鼠中枢及外周血淋巴细胞亚群重建的影响

目的 观察粒细胞集落刺激因子(G-CSF)对急性辐射损伤小鼠中枢及外周血淋巴细胞亚群重建的影响.方法 雌性BALB/c小鼠60只经6 Gy照射后随机分为照射组、G-CSF+照射组.G-CSF+照射组小鼠给与重组人G-CSF 100μg·kg-1·d-1皮下注射,连续14 d,照射组小鼠给与等体积磷酸盐缓冲液(PBS)皮下注射,连续14 d,另设空白对照组小鼠20只.照后7、14、21和28d颈部脱臼处死小鼠,取出胸腺制成单个核细胞悬液,使用流式细胞仪检测胸腺CD4+CD8+、CD4+CD8-、CD4-CD8+、CD4-CD8-细胞亚群的比例.使用全血细胞计数仪进行外周血白细胞计数及淋巴细胞绝对值测定,流式细胞仪检测照后14、28、60 d外周血淋巴细胞亚群比例,CCK-8法检测脂多糖(LPS)、刀豆蛋白A(ConA)刺激后脾脏淋巴细胞增殖指数.结果 照后7 d胸腺CD4+CD8+细胞比例降至最低,14 d出现反弹,21 d再次下降,以后逐渐恢复.照后28 d G-CSF+照射组CD4+CD8+细胞比例恢复正常并高于照射组(t=12.22,P＜0.05).照后21 d G-CSF+照射组CD4-CD8+细胞比例亦明显高于照射组(t=3.77,P＜0.05).照后7 d外周血白细胞及淋巴细胞绝对值降至最低,照后14和60 d,G-CSF+照射组CD3+CD8+T细胞比例明显高于照射组(t=4.31,5.78,P＜0.05),但两组间CD3+CD4+T细胞比例在各时间点无明显差异.G-CSF+照射组B淋巴细胞比例在照后14 d明显低于照射组(t=7.3,P＜0.05),但很快恢复,照后28和60 d两组B淋巴细胞比例差异无统计学意义.照后14 d,G-CSF+照射组脾脏淋巴细胞对LPS、ConA刺激的增殖指数分别为照射组的4.37和2.98倍.结论 G-CSF可促进照后胸腺细胞亚群的恢复,提高外周血淋巴细胞数量,调节外周血淋巴细胞亚群比例,提高淋巴细胞增殖功能,促进急性辐射损伤后中枢及外周免疫重建.

Abstract：

Objective To investigate the effects of recombinant human granulocyte colonystimulating factor(G-CSF) on central and peripheral lymphocyte subset reconstitution after a sublethal dose of irradiation. Methods Sixty female BALB/c mice were given a 6.0 Gy γ-ray total body irradiation (TBI) and randomly divided into 2 equal groups. The mice in G-CSF + TBI group were injected subcutaneously with recombinant human G-CSF 100 μg·kg-1·d-1 for 14 d and the mice in TBI group were injected subcutaneously with the same volume of phosphate buffered solution (PBS) once daily for 14 d. 7,14,21, and 28 d later the mice were killed and their thymus were taken out to prepare of the mononuclear cell suspension to analysis the percentage of thymic CD4 + CD8 + double positive, CD4 +CD8 - single positive, CD4 - CD8 + single positive and CD4 - CD8 - double negtive cells by flow cytometry. Peripheral blood samples were collected from the caudal vein twice a week, and the white blood cell(WBC) counts and absolute number of lymphocytes were assessed by automatic hemocyte analyzer. 14,28, and 60 d later blood samples were collected from angular vein to examine the peripheral lymphocyte subsets by flow cytometry. Cell counting kit-8 was used to detect lipopolysaccharide (LPS) or concanavalin A (ConA) stimulated splenic lymphocyte proliferation. Results The percentage of thymic CD4 + CD8 +double positive cells decreased 7 d after irradiation, rebounded at 14 d, decreased again at 21 d, and then got a permanent recovery. 28 d after irradiation the percentage of thymic CD4 + CD8 + double positive cells in the G-CSF + TBI group recovered to normal and was significantly higher than that of the TBI group (t =12. 22, P < 0. 05). 21d after irradiation the percentage of thymic CD4-CD8 + single positive cells of the G-CSF + TBI group was significantly higher than that of the TBI group (t = 3.77, P < 0. 05). The peripheral WBCs and lymphocytes decreased to the lowest levels 7 d after irradiation and then gradually increased, however, WBCs and lymphoeytes of the G-CSF + TBI group began to recover earlier and faster than the TBI group. The proportion of CD3 + CD8 + T cells of the G-CSF + TBI group was significantly higher than that of the TBI group 14 and 60 d after irradiation (t =4. 31,5.78, P <0.05). But there was no significant difference in the proportion of CD3 + CD4 + T cells between the two groups. The proportion of B lymphoeytes of the G-CSF + TBI group was significantly lower than that of the TBI group 14 d after irradiation(t =7.30, P <0.05), but it recovered quickly, and there were no significant differences in the proportion of B lymphoeytes between the two groups 28 and 60 d after irradiation. The proliferation indexes of splenic lymphocytes in response to LPS and ConA in the G-CSF + TBI group were 4. 37 and 2.98 times higher than those in the TBI group 14 d after irradiation. Conclusions G-CSF could accelerate the recovery of central and peripheral lymphocyte subsets, raise the absolute number of lymphocytes, and enhance their proliferative function, which contributes to the central and peripheral immune reconstitution after acute irradiation.     

HLA单倍型相合外周血干细胞联合间充质干细胞共移植治疗儿童重型再生障碍性贫血一例

患者,女,6岁.因发热伴咳嗽、咯痰8d,于2009年12月3日人院.查体:体温38.6℃,脉率120次/min,呼吸20次/min,血压70/40 mm Hg(1 mm Hg =0.133 kPa).神志清,精神可,重度贫血貌,全身皮肤散在出血点,巩膜无黄染,全身浅表淋巴结无明显肿大.咽部充血,扁桃体无肿大.心率120次/min,律齐.双肺呼吸音减低,右肺中下部可闻及湿性哕音.腹软,肝、脾肋缘下未触及.血常规:WBC 0.66×109/L,RBC 1.82×1012/L,HGB 56 g/L,PLT 0×109/L,网织红细胞占0.0022.胸骨、髂骨骨髓象:增生重度减低,粒系0,红系占0.06,淋巴细胞占0.855,巨核细胞未见,可见大量非造血细胞团.胸部CT:双肺弥漫性炎性病变阳性,以右肺为主.血培养:肺炎链球菌阳性,对万古霉素敏感.     

辐射生物剂量计的临床应用与研究现状

简述了现行的辐射生物剂量计在临床应用中的优点和局限性,以及目前的研究进展.对近年来较受关注的可能成为生物剂量计的新指标和技术方法作一简介,并对其可行性和潜在应用价值进行了探讨.     

山东济宁60Co辐射事故受照人员的心肌酶变化

山东省济宁市某辐照厂2名工作人员于2004年10月21日受到放射源60Co意外照射,病例A受照剂量为20～25Gy,病例B受照剂量为9～15Gy。分别诊断为轻度肠型急性放射病和极重度骨髓型急性放射病。借助心肌酶谱活性的动态变化,可以了解心肌损伤程度、心功能恢复情况及预后[1,2]。笔者对2     

原始骨髓间充质干细胞治疗慢性移植物抗宿主病临床研究方案

正慢性移植物抗宿主病(chronic graft versus host disease,c GVHD)是发生于异基因造血干细胞移植100 d后重要并发症,发生率为30%~60%。c GVHD是表现为类似自身免疫及其他免疫疾病的多系统损害的一种全身性疾病,可累及皮肤、肝脏、肾脏、周围神经等多脏器,造成患者生活质量严重下降,是移植晚期导致死亡的主要原因[1-3]。其发病机制尚     

过继性细胞免疫治疗白血病研究进展

细胞免疫治疗在消除白血病微小残留病和延长治疗后缓解期方面的作用正日益受到人们的重视.过继性回输免疫效应细胞是其中的重要方法之一.从20世纪80年代开始,人们一直在探索新的免疫活性细胞,期望得到更有效的治疗方法.本文综述了过继性细胞免疫治疗白血病的研究进展.     

异基因外周造血干细胞移植治疗极重度骨髓型急性放射病一例

目的探讨异基因外周血造血干细胞移植治疗极重度骨髓型急性放射病的效果，积累临床经验。方法1例患者意外受60^Co照射，受照射剂量为9～15Gy，诊断为极重度骨髓型急性放射病，在受照射后第4天患者开始接受预处理，3d后进行了HLA全相合的异基因外周造血干细胞移植。移植后采用环孢素A和霉酚酸酯预防移植物抗宿主病（GVHD）。结果造血功能于移植后第9天开始恢复，第11天WBC升至14．74×10^9L，随后降至正常范围，血小板升至50×10^9／L，Hb在80g／L以上。经短串联重复序列聚合酶链反应及血型动态检测，证实供者细胞稳定植入，原有染色体畸变和微核均消失，血型于移植后第27天完全转变为供者型。患者未发生GVHD，但放射性损伤持续加重，并发多重感染，于移植后第68天（受照射后第75天）死于多脏器功能衰竭。结论极重度骨髓型急性放射病可以通过异基因造血干细胞移植恢复造血功能，为患者存活创造机会，但仅有造血功能恢复，而未能解决好放射线对全身组织的损伤及免疫功能重建，患者仍难长期存活。