严重创伤对小鼠骨髓来源的树突状细胞诱导T细胞应答能力的影响
发布时间：2018-08-01 11:49
【摘要】： 目的:创伤是对人类生命健康与社会生产的严重威胁之一,在我国已成为继心脑血管疾病和恶性肿瘤之外最大的死亡原因。严重创伤后机体免疫功能紊乱可导致创伤后感染和器官功能障碍等并发症,深入研究有助于我们进一步了解创伤及其并发症的机制,并指导临床治疗。长期以来,针对创伤后免疫功能紊乱的研究涉及了中性粒细胞、单核-巨噬细胞和T、B淋巴细胞等诸多方面,但作为免疫系统重要的成员之一——树突状细胞(dendritic cell, DC)在创伤后免疫功能紊乱中的作用和地位迄今未被阐明。DC是联系先天性免疫和适应性免疫的桥梁,作为专职的抗原提呈细胞(APC),DC在启动和调节先天性及适应性免疫应答中具有关键性的作用。已有研究证实,机体遭遇创伤后,DC在其分化发育过程、抗原提呈功能和细胞因子产生等方面均发生了障碍。但作为中枢免疫器官的骨髓,在创伤后向DC分化的过程及骨髓来源DC(BMDC)的功能是否会发生改变,尚未见报道。本研究着重探讨了创伤对小鼠BMDC刺激T细胞增殖能力的影响。 方法:①制备动物创伤模型,致伤后24 h分离小鼠骨髓细胞,体外应用重组小鼠粒细胞/巨噬细胞—集落刺激因子(rmGM-CSF)诱导树突状细胞;②通过混合淋巴细胞反应(MLR)检测未成熟、成熟DC诱导异源T细胞的应答能力;③应用流式细胞术检测BMDC表面主要组织相容性复合物II类分子(MHC II)及共刺激分子CD40、CD80、CD86表达;④应用酶联免疫吸附试验(ELISA)检测脂多糖(LPS)刺激的BMDC培养上清中白细胞介素-12(IL-12)p40、IL-12 p70以及白细胞介素-10(IL-10)水平的变化。 结果:①在同等诱导条件下,创伤组小鼠骨髓细胞在体外诱导的DC(CD11c阳性细胞)百分率与正常对照组相比无明显差异,但其总数量明显低于正常对照组(骨髓细胞诱导DC的扩增倍数分别为3.9±0.5 vs 5.4±0.6,P 0.05);②创伤组小鼠BMDC,在经过LPS诱导成熟前后,其介导的MLR值均明显低于相应的对照组值(P 0.05);③以CD11c阳性细胞圈门,创伤小鼠BMDC在LPS刺激前后的CD40表达均明显低于正常对照组(4±1.0% vs 22±3.5%,P 0.01;56±7.5% vs 91±8.0%,P 0.01),但MHC II、CD80和CD86表达在两组间无统计学差异(P 0.05);④创伤组小鼠BMDC在体外经LPS诱导成熟后,其IL-12 p40、IL-12 p70分泌水平均明显低于正常对照组(45±6.5 ng/L vs 78±6.8 ng/L,P 0.05;9±1.0 ng/L vs 18±1.9 ng/L,P 0.05),但分泌IL-10的能力与正常对照组相比无统计学差异(P 0.05)。两组未成熟DC分泌上述细胞因子水平均无统计学差异(P 0.05) 结论:①创伤小鼠骨髓细胞对GM-CSF刺激增殖的敏感性降低;②创伤小鼠的BMDC功能发生障碍,其介导的混合淋巴细胞反应显著降低,对T细胞增殖的激发能力受到抑制;③创伤小鼠BMDC诱导T细胞应答的能力降低可能与其CD40表达及IL-12分泌减少有关。
[Abstract]:Objective: trauma is one of the serious threats to human life, health and social production. In our country, it has become the biggest cause of death outside the cardiovascular and cerebrovascular diseases and malignant tumors. The immune dysfunction of the body after severe trauma can lead to complications such as post traumatic infection and organ dysfunction. Further research will help us to further understand the creation of the disease. The mechanism of injury and its complications and guiding clinical treatment. For a long time, the study of post-traumatic immune dysfunction involves neutrophils, mononuclear macrophages, T, B lymphocytes and many other aspects, but as one of the important members of the immune system, the dendritic cell (DC) is in the post-traumatic immune dysfunction. The role and status have not been elucidated so far that.DC is a bridge linking innate and adaptive immunity. As a full-time antigen presenting cell (APC), DC plays a key role in the initiation and regulation of congenital and adaptive immune responses. It has been proved that after the body suffers injury, the DC is in its differentiation and development and the antigen presentation function However, there have been obstacles in the production of cytokine, but it has not been reported whether the bone marrow, as a central immune organ, can differentiate into DC after trauma and the function of bone marrow DC (BMDC). This study focuses on the effect of trauma on the proliferation of T cells stimulated by BMDC in mice.
Methods: (1) the animal model of trauma was prepared. The bone marrow cells of mice were separated by 24 h after injury. The recombinant mouse granulocyte / macrophage colony stimulating factor (rmGM-CSF) was used to induce dendritic cells in vitro. (2) the response ability of immature T cells induced by mature DC was detected by mixed lymphocyte reaction (MLR); (3) flow cytometry was used to detect the cells. The main histocompatibility complex II molecules (MHC II) and co stimulator CD40, CD80, CD86 expression on the BMDC surface; (4) the enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin -12 (IL-12) and interleukin (IL) in the culture supernatant of BMDC culture of LPS (LPS).
Results: (1) there was no significant difference in the percentage of DC (CD11c positive cells) induced by bone marrow cells in the trauma group in vitro compared with the normal control group, but the total number of bone marrow cells was significantly lower than that of the normal control group (3.9 + 0.5 vs 5.4 + 0.6, P 0.05) of the bone marrow cells induced by bone marrow cells (P 0.05). (2) the BMDC in the trauma group was in L The MLR value of PS was significantly lower than that of the corresponding control group (P 0.05) before and after the induction of maturation. (3) the expression of CD40 in the traumatic mice BMDC before and after LPS stimulation was significantly lower than that of the normal control group (4 + 1% vs 22 + 3.5%, P 0.01, 56 + 7.5% vs 91 + 8%, P 0.01), but there was no statistical difference between the two groups. Difference (P 0.05); (4) the level of IL-12 P40 and IL-12 p70 secretion in the IL-12 P40 and IL-12 p70 in the trauma group was significantly lower than that in the normal control group (45 + 6.5 ng/L vs 78 + 6.8 ng/L, P 0.05, 9 + 1 ng/L 18 + 1.9), but there was no statistical difference between the normal control group and the normal control group. There was no significant difference in secretion of cytokines mentioned above (P 0.05).
Conclusion: (1) the sensitivity of bone marrow cells in traumatized mice decreased to the proliferation of GM-CSF, and the BMDC function in traumatic mice was impaired, the mixed lymphocyte reaction was significantly reduced and the ability to stimulate the proliferation of T cells was inhibited. The ability to respond to BMDC induced T cells in traumatized mice may be associated with the expression of CD40 and the secretion of IL-12. The reduction is related.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
本文编号：2157482
[Abstract]:Objective: trauma is one of the serious threats to human life, health and social production. In our country, it has become the biggest cause of death outside the cardiovascular and cerebrovascular diseases and malignant tumors. The immune dysfunction of the body after severe trauma can lead to complications such as post traumatic infection and organ dysfunction. Further research will help us to further understand the creation of the disease. The mechanism of injury and its complications and guiding clinical treatment. For a long time, the study of post-traumatic immune dysfunction involves neutrophils, mononuclear macrophages, T, B lymphocytes and many other aspects, but as one of the important members of the immune system, the dendritic cell (DC) is in the post-traumatic immune dysfunction. The role and status have not been elucidated so far that.DC is a bridge linking innate and adaptive immunity. As a full-time antigen presenting cell (APC), DC plays a key role in the initiation and regulation of congenital and adaptive immune responses. It has been proved that after the body suffers injury, the DC is in its differentiation and development and the antigen presentation function However, there have been obstacles in the production of cytokine, but it has not been reported whether the bone marrow, as a central immune organ, can differentiate into DC after trauma and the function of bone marrow DC (BMDC). This study focuses on the effect of trauma on the proliferation of T cells stimulated by BMDC in mice.
Methods: (1) the animal model of trauma was prepared. The bone marrow cells of mice were separated by 24 h after injury. The recombinant mouse granulocyte / macrophage colony stimulating factor (rmGM-CSF) was used to induce dendritic cells in vitro. (2) the response ability of immature T cells induced by mature DC was detected by mixed lymphocyte reaction (MLR); (3) flow cytometry was used to detect the cells. The main histocompatibility complex II molecules (MHC II) and co stimulator CD40, CD80, CD86 expression on the BMDC surface; (4) the enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin -12 (IL-12) and interleukin (IL) in the culture supernatant of BMDC culture of LPS (LPS).
Results: (1) there was no significant difference in the percentage of DC (CD11c positive cells) induced by bone marrow cells in the trauma group in vitro compared with the normal control group, but the total number of bone marrow cells was significantly lower than that of the normal control group (3.9 + 0.5 vs 5.4 + 0.6, P 0.05) of the bone marrow cells induced by bone marrow cells (P 0.05). (2) the BMDC in the trauma group was in L The MLR value of PS was significantly lower than that of the corresponding control group (P 0.05) before and after the induction of maturation. (3) the expression of CD40 in the traumatic mice BMDC before and after LPS stimulation was significantly lower than that of the normal control group (4 + 1% vs 22 + 3.5%, P 0.01, 56 + 7.5% vs 91 + 8%, P 0.01), but there was no statistical difference between the two groups. Difference (P 0.05); (4) the level of IL-12 P40 and IL-12 p70 secretion in the IL-12 P40 and IL-12 p70 in the trauma group was significantly lower than that in the normal control group (45 + 6.5 ng/L vs 78 + 6.8 ng/L, P 0.05, 9 + 1 ng/L 18 + 1.9), but there was no statistical difference between the normal control group and the normal control group. There was no significant difference in secretion of cytokines mentioned above (P 0.05).
Conclusion: (1) the sensitivity of bone marrow cells in traumatized mice decreased to the proliferation of GM-CSF, and the BMDC function in traumatic mice was impaired, the mixed lymphocyte reaction was significantly reduced and the ability to stimulate the proliferation of T cells was inhibited. The ability to respond to BMDC induced T cells in traumatized mice may be associated with the expression of CD40 and the secretion of IL-12. The reduction is related.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
【参考文献】
相关期刊论文 前4条
1 韩健;梁华平;;严重创伤对小鼠体内单核细胞向树突状细胞分化及迁移过程的影响[J];创伤外科杂志;2007年01期
2 郭靖;王震平;梁华平;谭立志;;失血合并闭合性骨折致小鼠脾脏树突状细胞抗原递呈功能下降[J];创伤外科杂志;2007年03期
3 白祥军,唐朝晖,邹声泉;严重多发伤患者外周血树突状细胞的变化[J];中华急诊医学杂志;2004年04期
4 王震平;梁华平;;树突状细胞在介导创伤后迟发型超敏反应受抑中的作用[J];中国医学科学院学报;2007年04期
，本文编号：2157482
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2157482.html
最近更新
教材专著
