摘要
背景:预输注供者凋亡细胞能够诱导受者体内产生针对供者抗原发生的特异性免疫耐受,但其具体途径尚不清楚。目的:观察供体凋亡淋巴细胞输注后受体外周血细胞因子的变化。设计、时间及地点:细胞观察性实验,于2007-11/2008-05在南方医科大学附属珠江医院移植免疫实验室完成。材料:供体C57BL雄性小鼠14只,8～10周龄,受体Balb/c雄性小鼠50只,6～8周龄。方法:分离供体小鼠脾脏细胞,放置于紫外线灯距离20cm处的摇床上照射1h,紫外线诱导C57BL小鼠脾脏细胞凋亡。将受体Balb/c小鼠随机分为凋亡细胞组和活细胞组,分别将107个供体C57小鼠脾脏凋亡细胞、活细胞经尾静脉输注于受体小鼠。主要观察指标:输注后0,1,3,6,12h经心脏采血,Luminex技术液态芯片法检测外周血白细胞介素1β、白细胞介素2、白细胞介素4、白细胞介素5、白细胞介素6、白细胞介素10、白细胞介素12、粒细胞-巨细胞集落刺激因子、干扰素γ、肿瘤坏死因子变化。结果:只检测到白细胞介素6、白细胞介素10、白细胞介素12、粒细胞-巨细胞集落刺激因子、干扰素γ5种细胞因子,其他细胞因子未测出。经紫外线诱导后小鼠脾脏细胞凋亡率达到93.86%。与0h相比,外周血中干扰素γ和白细胞介素10分别在输注活细胞后6h和3,6h明显升高,其他细胞因子无明显变化。与0h相比,输注凋亡细胞1,3,6,12h后,外周血中各细胞因子无明显变化。活细胞组输注后1h白细胞介素6和输注后3,6h白细胞介素10浓度高于凋亡细胞组,其他细胞因子差异不显著。结论:同种异源供者凋亡淋巴细胞输注对外周血细胞因子浓度无变化,提示其对受体外周免疫功能可能无明显影响,其诱导产生的免疫耐受可能主要在局部发挥作用。
BACKGROUND： The pre-infusion of allogeneic donor apoptotic cell can induce special immunological tolerance aimed at donor antigens, but the concrete ways are unclear.
OBJECTIVE： To observe the changes of cytokines in recipient＇s peripheral blood after donor apoptotic lymphocyte infusion. DESIGN, TIME AND SETTING： A cellular observational experiment was performed at Transplantation Immunity Laboratory in Affiliated Zhujiang Hospital of Southern Medical University from November 2007 to May 2008.
MATERIALS： Fourteen C57BL male mice of 8 to 10weeks old were assigned as donors, and fifty Balb/C male mice of 6 to 8 weeks old were assigned as recipients.
METHODS： Spleen cells were separated from donor rats, and then placed on the table concentrator 20 cm away from an ultraviolet lamp for 1 hour in order to induce apoptosis of spleen cells. Balb/c mice were randomly divided into 2 groups： apoptotic cell and living cell groups, with 25 rats in each group. Each mouse in the apoptotic cell and living cell groups was infused with 107 apoptotic cells or living cells, respectively.
MAIN OUTCOME MEASURES： Blood was collected via heart puncture at 0, 1,3, 6, and 12 hours after infusion. The changes of interleukin-1 β, interleukin-2, interleukin-4, interleukin-5, interleukin-6, interleukin-10, interleukin-12, granulocyte macrophage colony stimulating factor, interferon-γ, and tumor necrosis factor in peripheral blood were observed by Luminex. RESULTS： The interleukin-6, interleukin-10, interleukin-12, granulocyte macrophage colony stimulating factor, and interferon-y were detected, but other cytokines could not detect. The apoptotic rate of spleen cells after UV treatment was 93%. In the living cell group, the concentration of interferon-γ at 6 hours after transfusion and concentration of interleukin-10 at 3 and 6 hours significantly increased compared to those at 0 hour, respectively. And other cytokines had no obvious change. In the apoptotic cell group, all cytokines had no obvious change at 1,3, 6
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第18期3442-3446,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家教育部器官移植重点实验室开放基金
国家自然科学基金面上项目资助(30571757)
广东省自然科学基金重点项目资助(037055)~~
