不同免疫状态对小鼠脊髓损伤后修复的影响
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摘要
免疫炎症反应是机体免疫系统抵御外界环境和维护自身的正常生理过程。而当组织损伤时,它也是重要参与者,既可抵抗损伤处感染,又可为损伤组织提供细胞因子利于其再生修复。但免疫炎症反应在脊髓损伤后的作用备受争议:一方面,脊髓损伤后激活的免疫炎症反应造成脊髓组织继发性损伤恶化;另一方面,免疫炎症反应清除细胞碎片并分泌对神经元起保护作用的细胞因子如转化生长因子-β(TGF-β)、脑源性神经营养因子(BDNF)、神经营养因子-3(NT-3)和神经生长因子(NGF)等,利于损伤脊髓组织的修复和再生。
但是笼统的说脊髓损伤后的自身免疫炎症反应好与坏是不科学的,因为免疫炎症细胞需要相互联系和影响,形成行使作用的免疫细胞网络从而发挥免疫功能。即形成特定的免疫状态,发挥特殊作用。不同免疫状态下,自身免疫反应有很大的差别。
我们前期实验发现,脊髓损伤后的免疫炎症反应具有双面性,在脊髓损伤早期,它们主要表现为破坏性作用,加重脊髓损伤;而时间进程的推进,又逐渐转化为神经保护性作用,促进损伤组织的修复。这使我们想到是不是损伤区的免疫状态发生了转变,致使免疫炎症反应的作用也发生了转型呢?如果的确存在转变,那么又是那些类型的细胞起了关键性作用呢?
为了证实我们的假设,我们试图通过分析不同免疫状态对小鼠脊髓损伤修复的影响,寻找更利于脊髓损伤修复的免疫状态、不同免疫状态下损伤区的差异和发挥关键作用的细胞类型。为探索通过干预免疫反应过程来治疗脊髓损伤的治疗策略提供更有力的理论基础。
实验中用到三种免疫状态下小鼠,即BALB/c、DO11.10小鼠和无胸腺裸鼠。其中BALB/c小鼠为正常免疫状态小鼠,DO11.10小鼠和无胸腺裸鼠均为BALB/c遗传背景下外周分别缺失正常功能CD4+ T淋巴细胞和T淋巴
细胞的小鼠。实验分为两部分:
1.不同免疫状态小鼠脊髓损伤后运动功能恢复状况的差异
通过SCI后11 w的BBB评分和足迹分析两种行为学实验来评价三种免疫状态小鼠运动功能恢复状况。
2.不同免疫状态小鼠脊髓损伤区的细胞学和形态学差异
通过SCI后1 w、2 w和3 w三个时间点的H&E染色、巨噬细胞免疫荧光三标记实验和T淋巴细胞的三标记实验,来分析三种免疫状态小鼠脊髓损伤区免疫细胞的差异。损伤区域残存神经纤维160(NF160)和胶质原纤维酸性蛋白(GFAP)的免疫荧光实验,观察三种小鼠脊髓损伤区局部形态学差异。通过以上两部分实验,我们得到了如下的实验结果:
1.三种免疫状态对小鼠的脊髓损伤修复有明显的影响
首先,从BBB评分来分析运动功能恢复情况:无胸腺裸鼠一直明显好于BALB/c和DO11.10小鼠(p<0.01);DO11.10小鼠在损伤后4 w内要好于持续低迷的BALB/c小鼠(p<0.05),但4 w后BALB/c小鼠恢复较快,到6 w时几乎追平了DO11.10小鼠,而到7 w后已经远好于DO11.10小鼠(p<0.01)。
足迹分析实验也证实了三种免疫状态小鼠脊髓损伤恢复的差异。SCI后2 w,BALB/c小鼠后肢状况仍很差,基本无运动能恢复;DO11.10小鼠稍好,已有明显的后足运动的趋势和成一定频率的左右侧互换;裸鼠要好于前两者,首先裸鼠足底可以着地,尽管不那么恒定,其次,可以看到不太稳定的左右协调运动。到3 w时,三组小鼠都出现了相应的进步。BALB/c组可看到一定的步履,尽管主要靠足背侧产生,偶尔会出现足底着地;而DO11.10小鼠则出现了一侧足底着地和一定的协调性的现象;裸鼠依然是最稳健的,基本全是足底着地,协调性也好于2 w时的状况。6 w和11 w的步长和步间距统计分析结果基本与BBB评分结果相一致,裸鼠状态明显好于BALB/c和DO11.10组(p<0.01)。11 w时BALB/c小鼠稍好于DO11.10小鼠,但统计分析无显著差异。
2.不同免疫状态小鼠脊髓损伤区免疫细胞的存在明显差异
损伤1 w的H&E染色显示DO11.10小鼠和裸鼠损伤区免疫炎性细胞稍多于BALB/c小鼠。巨噬细胞免疫荧光三标记的实验显示,DO11.10小鼠和裸鼠损伤区M2型巨噬细胞比BALB/c小鼠多,裸鼠损伤区及周边CD11b阳性巨噬/小胶质细胞要比BALB/c和DO11.10小鼠多。M1型巨噬细胞无明显差异。T淋巴细胞三标记的免疫荧光实验结果则证实,三种小鼠免疫状态有很大的差别。3 w时,BALB/c小鼠脊髓损伤区已出现大量活化的CD4+、CD8+ T淋巴细胞;DO11.10小鼠损伤区仅存在个别CD4+ T淋巴细胞,但处于无活性状态;裸鼠则基本无T淋巴细胞出现在损伤区及周边。
2 w时, NF160和GFAP免疫荧光实验也证实,三种免疫状态小鼠脊髓损伤区局部存在形态学差异。GFAP免疫荧光实验显示DO11.10小鼠和裸鼠损伤边界较清晰。而NF160阳性神经纤维则在裸鼠脊髓损伤区存在较多。
通过以上两部分实验,我们认为BALB/c、DO11.10小鼠和无胸腺裸鼠三种不同的免疫状态的确对其自身脊髓损伤恢复状态产生了很大的影响。裸鼠的免疫状态更利于其脊髓损伤修复。分析三种免疫状态下脊髓损伤区免疫细胞我们发现,精氨酸酶Ⅰ、CD11b阳性巨噬细胞的较高表达和T淋巴细胞缺失的免疫状态可能更较利于脊髓损伤后的修复。而BALB/c小鼠特殊的恢复状况则提示我们,T淋巴细胞功能的复杂性与特定时期的免疫状态有相关性。
The role of immunoreactions is against the external environment and maintaining its normal physiological function. When tissue is injured, it not only takes part in resisting infection, but also helping the regeneration and repair of the injured tissue through secreting cytokines. But the immunoreactions have controversial role after spinal cord injury: On the one hand, inflammatory response activated by spinal cord injury(SCI) may worsen the secondary injury; On the other hand, the immune response can clear the cell debris and secret several cytokines such as transforming growth factor-β(TGF-β), brain-derived neurotrophic factor (BDNF), neurotrophin -3 (NT-3) and nerve growth factor (NGF), etc, that can protect neurons from injury and help spinal cord injury repairing and regenerating.
It is not suitable to that the autoimmunity inflammation after spinal cord injury is good or bad. Because immune and inflammatory cells can contact mutually and influence the formation of networks, and then exert the role of immune cells. And then formatting those special immune statuses, that plays a distinctive role. Different immune states may have very different immune responses.
Our previous experiments showed that the immune inflammatory response may have double faces in spinal cord injury. Early after spinal cord injury, they mainly display their destructive role and worsen spinal cord injury; and at latter time, course of the advance, they gradually transformed into neural protective role and promoting repair of injured tissues. This leads us to wonder whether the injured area has changed its immune state, resulting in the role of immune and inflammatory response transformed into another condition. If the change does exist, and then which types of cells play a key role?
To confirm the assumption that we put forward, we tried to analysis the mice’s locomotion recovery with different immune status after spinal cord injury, looking for advanced understanding of the various immune status’influence on spinal cord injury repair. And then we can distinguish their different and hunt for the better immune status for spinal cord injury repair and the crucial cells or molecular. We are aim to explore the intervention strategies by tune the immune response for a better treatment after spinal cord injury.
In our experiment, we used three kinds of immune status mice. Those are BALB/c, DO11.10 and nude mice. As we known, BALB/c mice have normal immune, while DO11.10 mice and nude mice are transgenic mice of BALB/c genetic background. DO11.10 mice are missing the normal function of peripheral CD4+ T lymphocytes and nude mice are athymic and lack of T lymphocytes. Our experiment was divided into two parts:
1. The different functional recovery between the mice with different immune situation after spinal cord injury. We evaluate the recovery of locomotion function behavior of mice with different immune state in mice by the BBB score and footprint experiments through a long time as 11 w after spinal cord injury.
2. The cellular and morphological differences between the mice with different immune situation after spinal cord injury.
First, at the time points of 1 w, 2 w and 3 w after SCI, we use H & E staining, immunofluorescence staining of macrophages and T lymphocytes in three labeling experiments to analyze the immune cells difference of the spinal cord injury areas in the different immune status mice.
And then we used residual nerve fiber 160 (NF160) and glial fibrillary acidic protein (GFAP) immunofluorescence experiments to compare the three kinds of mice’morphological differences of focal spinal cord damage region.
We get the following results by the above two parts experiments:
1. Three kinds of immune status have significant impact on locomotions recovery after spinal cord injury in mice.
First of all, from the BBB scoreing result, we can see the three different mice’recovery of motor function as follow: Nude mice are significantly better than BALB/c and DO11.10 mice (p <0.01); Before 4 weeks after injury, DO11.10 mice are better than the of BALB/c mice (p <0.05), but after 4 weeks BALB / c mice recovered rapidly. What’s more at 6 w BALB/c and DO11.10 mice are almost tied and at the 7 w BALB/c mice already much better than DO11. 10 mice(p <0.01).
Footprint experiments also confirmed that the three different immune statuses play a important role on spinal cord injury recovery of mice. 2 w after SCI, BALB / c mice’hindlimb position is still very poor, and almost no movement of any jonit; DO11.10 mice were slightly better, and there are obvious trends to move and a certain frequency of the leftside and rightside exchanges; Although Nude mice are less constant, but they are much more better than BALB/c and DO11.10. What’s more, the nude mice already have leftside and rightside coordinated motion unsteadily. At the 3 w time point, those three groups of mice have shown their corresponding improvements. BALB/c group has some certain of walking, even though it is mainly produced by the dorsal foot with occasional foot landing; DO11.10 mice appears one side plantar landing and some certain of coordination; Nude mice remains the most robust, with basically using all the foot landing, and its coordination are much better than 2 w’s situation. The statistical analysis of the step and step spacing at 6 w and 11 w is basically consistent with the BBB score. Nude mice was obviously better than the BALB/c and DO11.10 group (p <0.01). At 11 w time point, BALB/c mice are slightly better than DO11.10 mice, but no significant difference in statistical analysis.
2. The mice with different immune status have significantly different immune cells in the spinal cord injury areas.
1 w after injury, H & E staining showed that DO11.10 mice and nude mice have more inflammatory cells than BALB/c mice in the damage zone and approaching area. Macrophages immunofluorescence experiments showed that there are more M2 type macrophages in DO11.10 mice and nude mice damage zone than in BALB/c mice(P<0.01). And nude mice have much more CD11b-positive macrophages / microglia in the damage zone and surrounding than BALB/c and DO11.10 mice(P<0.01). M1 type macrophages are no significant difference in BALB/c and DO11.10 mice. T lymphocytes in the three labeled immunofluorescence experiments confirmed that the mice with different immune status are quite different in the T lymphocyte number. At 3 w time point, BALB / c mice have a large number of activated CD4+, CD8+ T lymphocytes in spinal cord injury area; DO11.10 mice’damage zone exists only individual CD4+ T lymphocytes, but no activity in the state; Nude mice are almost no T lymphocytes appearing in the damage zone and the surrounding.
2 w after SCI, NF160 and GFAP immunofluorescence experiments confirmed that those three groups with immune status exist morphological differences in the areas of local spinal cord injury. GFAP immunofluorescence experiments showed that DO11.10 mice and Nude mice damage boundaries clearer. And NF160 positive nerve fibers in the spinal cord injury area of nude mice are much larger.
With the above two experiments, we supose that those three different immune statuses of BALB/c, DO11.10 and athymic nude mice have a great impact on the locomotion recovery from their spinal cord injury. The immune status of nude mice is more conducive to their recovery from spinal cord injury. We found that the high level of arginaseⅠ, CD11b positive macrophages and T lymphocytes expression in immune deficiency state may be more beneficial for the repair of injuried spinal cord through analysis of three kinds of immune immune cells in the spinal cord injury area. We conclude that the complex function of T lymphocyte and the particular immune status is very relevant with the special recovery from spinal cord injury of BALB/c mice.
但是笼统的说脊髓损伤后的自身免疫炎症反应好与坏是不科学的,因为免疫炎症细胞需要相互联系和影响,形成行使作用的免疫细胞网络从而发挥免疫功能。即形成特定的免疫状态,发挥特殊作用。不同免疫状态下,自身免疫反应有很大的差别。
我们前期实验发现,脊髓损伤后的免疫炎症反应具有双面性,在脊髓损伤早期,它们主要表现为破坏性作用,加重脊髓损伤;而时间进程的推进,又逐渐转化为神经保护性作用,促进损伤组织的修复。这使我们想到是不是损伤区的免疫状态发生了转变,致使免疫炎症反应的作用也发生了转型呢?如果的确存在转变,那么又是那些类型的细胞起了关键性作用呢?
为了证实我们的假设,我们试图通过分析不同免疫状态对小鼠脊髓损伤修复的影响,寻找更利于脊髓损伤修复的免疫状态、不同免疫状态下损伤区的差异和发挥关键作用的细胞类型。为探索通过干预免疫反应过程来治疗脊髓损伤的治疗策略提供更有力的理论基础。
实验中用到三种免疫状态下小鼠,即BALB/c、DO11.10小鼠和无胸腺裸鼠。其中BALB/c小鼠为正常免疫状态小鼠,DO11.10小鼠和无胸腺裸鼠均为BALB/c遗传背景下外周分别缺失正常功能CD4+ T淋巴细胞和T淋巴
细胞的小鼠。实验分为两部分:
1.不同免疫状态小鼠脊髓损伤后运动功能恢复状况的差异
通过SCI后11 w的BBB评分和足迹分析两种行为学实验来评价三种免疫状态小鼠运动功能恢复状况。
2.不同免疫状态小鼠脊髓损伤区的细胞学和形态学差异
通过SCI后1 w、2 w和3 w三个时间点的H&E染色、巨噬细胞免疫荧光三标记实验和T淋巴细胞的三标记实验,来分析三种免疫状态小鼠脊髓损伤区免疫细胞的差异。损伤区域残存神经纤维160(NF160)和胶质原纤维酸性蛋白(GFAP)的免疫荧光实验,观察三种小鼠脊髓损伤区局部形态学差异。通过以上两部分实验,我们得到了如下的实验结果:
1.三种免疫状态对小鼠的脊髓损伤修复有明显的影响
首先,从BBB评分来分析运动功能恢复情况:无胸腺裸鼠一直明显好于BALB/c和DO11.10小鼠(p<0.01);DO11.10小鼠在损伤后4 w内要好于持续低迷的BALB/c小鼠(p<0.05),但4 w后BALB/c小鼠恢复较快,到6 w时几乎追平了DO11.10小鼠,而到7 w后已经远好于DO11.10小鼠(p<0.01)。
足迹分析实验也证实了三种免疫状态小鼠脊髓损伤恢复的差异。SCI后2 w,BALB/c小鼠后肢状况仍很差,基本无运动能恢复;DO11.10小鼠稍好,已有明显的后足运动的趋势和成一定频率的左右侧互换;裸鼠要好于前两者,首先裸鼠足底可以着地,尽管不那么恒定,其次,可以看到不太稳定的左右协调运动。到3 w时,三组小鼠都出现了相应的进步。BALB/c组可看到一定的步履,尽管主要靠足背侧产生,偶尔会出现足底着地;而DO11.10小鼠则出现了一侧足底着地和一定的协调性的现象;裸鼠依然是最稳健的,基本全是足底着地,协调性也好于2 w时的状况。6 w和11 w的步长和步间距统计分析结果基本与BBB评分结果相一致,裸鼠状态明显好于BALB/c和DO11.10组(p<0.01)。11 w时BALB/c小鼠稍好于DO11.10小鼠,但统计分析无显著差异。
2.不同免疫状态小鼠脊髓损伤区免疫细胞的存在明显差异
损伤1 w的H&E染色显示DO11.10小鼠和裸鼠损伤区免疫炎性细胞稍多于BALB/c小鼠。巨噬细胞免疫荧光三标记的实验显示,DO11.10小鼠和裸鼠损伤区M2型巨噬细胞比BALB/c小鼠多,裸鼠损伤区及周边CD11b阳性巨噬/小胶质细胞要比BALB/c和DO11.10小鼠多。M1型巨噬细胞无明显差异。T淋巴细胞三标记的免疫荧光实验结果则证实,三种小鼠免疫状态有很大的差别。3 w时,BALB/c小鼠脊髓损伤区已出现大量活化的CD4+、CD8+ T淋巴细胞;DO11.10小鼠损伤区仅存在个别CD4+ T淋巴细胞,但处于无活性状态;裸鼠则基本无T淋巴细胞出现在损伤区及周边。
2 w时, NF160和GFAP免疫荧光实验也证实,三种免疫状态小鼠脊髓损伤区局部存在形态学差异。GFAP免疫荧光实验显示DO11.10小鼠和裸鼠损伤边界较清晰。而NF160阳性神经纤维则在裸鼠脊髓损伤区存在较多。
通过以上两部分实验,我们认为BALB/c、DO11.10小鼠和无胸腺裸鼠三种不同的免疫状态的确对其自身脊髓损伤恢复状态产生了很大的影响。裸鼠的免疫状态更利于其脊髓损伤修复。分析三种免疫状态下脊髓损伤区免疫细胞我们发现,精氨酸酶Ⅰ、CD11b阳性巨噬细胞的较高表达和T淋巴细胞缺失的免疫状态可能更较利于脊髓损伤后的修复。而BALB/c小鼠特殊的恢复状况则提示我们,T淋巴细胞功能的复杂性与特定时期的免疫状态有相关性。
The role of immunoreactions is against the external environment and maintaining its normal physiological function. When tissue is injured, it not only takes part in resisting infection, but also helping the regeneration and repair of the injured tissue through secreting cytokines. But the immunoreactions have controversial role after spinal cord injury: On the one hand, inflammatory response activated by spinal cord injury(SCI) may worsen the secondary injury; On the other hand, the immune response can clear the cell debris and secret several cytokines such as transforming growth factor-β(TGF-β), brain-derived neurotrophic factor (BDNF), neurotrophin -3 (NT-3) and nerve growth factor (NGF), etc, that can protect neurons from injury and help spinal cord injury repairing and regenerating.
It is not suitable to that the autoimmunity inflammation after spinal cord injury is good or bad. Because immune and inflammatory cells can contact mutually and influence the formation of networks, and then exert the role of immune cells. And then formatting those special immune statuses, that plays a distinctive role. Different immune states may have very different immune responses.
Our previous experiments showed that the immune inflammatory response may have double faces in spinal cord injury. Early after spinal cord injury, they mainly display their destructive role and worsen spinal cord injury; and at latter time, course of the advance, they gradually transformed into neural protective role and promoting repair of injured tissues. This leads us to wonder whether the injured area has changed its immune state, resulting in the role of immune and inflammatory response transformed into another condition. If the change does exist, and then which types of cells play a key role?
To confirm the assumption that we put forward, we tried to analysis the mice’s locomotion recovery with different immune status after spinal cord injury, looking for advanced understanding of the various immune status’influence on spinal cord injury repair. And then we can distinguish their different and hunt for the better immune status for spinal cord injury repair and the crucial cells or molecular. We are aim to explore the intervention strategies by tune the immune response for a better treatment after spinal cord injury.
In our experiment, we used three kinds of immune status mice. Those are BALB/c, DO11.10 and nude mice. As we known, BALB/c mice have normal immune, while DO11.10 mice and nude mice are transgenic mice of BALB/c genetic background. DO11.10 mice are missing the normal function of peripheral CD4+ T lymphocytes and nude mice are athymic and lack of T lymphocytes. Our experiment was divided into two parts:
1. The different functional recovery between the mice with different immune situation after spinal cord injury. We evaluate the recovery of locomotion function behavior of mice with different immune state in mice by the BBB score and footprint experiments through a long time as 11 w after spinal cord injury.
2. The cellular and morphological differences between the mice with different immune situation after spinal cord injury.
First, at the time points of 1 w, 2 w and 3 w after SCI, we use H & E staining, immunofluorescence staining of macrophages and T lymphocytes in three labeling experiments to analyze the immune cells difference of the spinal cord injury areas in the different immune status mice.
And then we used residual nerve fiber 160 (NF160) and glial fibrillary acidic protein (GFAP) immunofluorescence experiments to compare the three kinds of mice’morphological differences of focal spinal cord damage region.
We get the following results by the above two parts experiments:
1. Three kinds of immune status have significant impact on locomotions recovery after spinal cord injury in mice.
First of all, from the BBB scoreing result, we can see the three different mice’recovery of motor function as follow: Nude mice are significantly better than BALB/c and DO11.10 mice (p <0.01); Before 4 weeks after injury, DO11.10 mice are better than the of BALB/c mice (p <0.05), but after 4 weeks BALB / c mice recovered rapidly. What’s more at 6 w BALB/c and DO11.10 mice are almost tied and at the 7 w BALB/c mice already much better than DO11. 10 mice(p <0.01).
Footprint experiments also confirmed that the three different immune statuses play a important role on spinal cord injury recovery of mice. 2 w after SCI, BALB / c mice’hindlimb position is still very poor, and almost no movement of any jonit; DO11.10 mice were slightly better, and there are obvious trends to move and a certain frequency of the leftside and rightside exchanges; Although Nude mice are less constant, but they are much more better than BALB/c and DO11.10. What’s more, the nude mice already have leftside and rightside coordinated motion unsteadily. At the 3 w time point, those three groups of mice have shown their corresponding improvements. BALB/c group has some certain of walking, even though it is mainly produced by the dorsal foot with occasional foot landing; DO11.10 mice appears one side plantar landing and some certain of coordination; Nude mice remains the most robust, with basically using all the foot landing, and its coordination are much better than 2 w’s situation. The statistical analysis of the step and step spacing at 6 w and 11 w is basically consistent with the BBB score. Nude mice was obviously better than the BALB/c and DO11.10 group (p <0.01). At 11 w time point, BALB/c mice are slightly better than DO11.10 mice, but no significant difference in statistical analysis.
2. The mice with different immune status have significantly different immune cells in the spinal cord injury areas.
1 w after injury, H & E staining showed that DO11.10 mice and nude mice have more inflammatory cells than BALB/c mice in the damage zone and approaching area. Macrophages immunofluorescence experiments showed that there are more M2 type macrophages in DO11.10 mice and nude mice damage zone than in BALB/c mice(P<0.01). And nude mice have much more CD11b-positive macrophages / microglia in the damage zone and surrounding than BALB/c and DO11.10 mice(P<0.01). M1 type macrophages are no significant difference in BALB/c and DO11.10 mice. T lymphocytes in the three labeled immunofluorescence experiments confirmed that the mice with different immune status are quite different in the T lymphocyte number. At 3 w time point, BALB / c mice have a large number of activated CD4+, CD8+ T lymphocytes in spinal cord injury area; DO11.10 mice’damage zone exists only individual CD4+ T lymphocytes, but no activity in the state; Nude mice are almost no T lymphocytes appearing in the damage zone and the surrounding.
2 w after SCI, NF160 and GFAP immunofluorescence experiments confirmed that those three groups with immune status exist morphological differences in the areas of local spinal cord injury. GFAP immunofluorescence experiments showed that DO11.10 mice and Nude mice damage boundaries clearer. And NF160 positive nerve fibers in the spinal cord injury area of nude mice are much larger.
With the above two experiments, we supose that those three different immune statuses of BALB/c, DO11.10 and athymic nude mice have a great impact on the locomotion recovery from their spinal cord injury. The immune status of nude mice is more conducive to their recovery from spinal cord injury. We found that the high level of arginaseⅠ, CD11b positive macrophages and T lymphocytes expression in immune deficiency state may be more beneficial for the repair of injuried spinal cord through analysis of three kinds of immune immune cells in the spinal cord injury area. We conclude that the complex function of T lymphocyte and the particular immune status is very relevant with the special recovery from spinal cord injury of BALB/c mice.
引文
吴俊芳,刘忞，等. M,《现代神经科学研究方法》.2005,723-725.
Anderson AJ, Robert S, et al. Activation of complement pathways after contusion-induced spinal cord injury. J. Neurotrauma. 2004; 21(12): 1831–1846.
Ankeny DP, Popovich PG, et al. Mechanisms and implications of adaptive immune responses after traumatic spinal cord injury. J. Neuroscience, 2009; 158(3): 1112–21.
Arumugam T, Woodruff TM, et al. Neuroprotection in stroke by complement inhibition and immunoglobulin therapy. J. Neuroscience. 2009; 158(3): 1074–1089.
Bakalash S, Shlomo GB, et al. T-cell based vaccination for morphological and functional neuroprotection in a rat model of chronically elevated intraocular pressure. J. Mol Med. 2005; 83(11): 904–916.
Barnum SR. Inhibition of complement as a therapeutic approach in inflammatory central nervous system (CNS) disease. J. Mol Med. 1999; 5(9): 569–582.
Basso DM, Beattie MS, Bresnahan JC. MASCIS evaluation of open field locomotor scores: Effects of Experience and Teamwork on Reliability. J. Neurotroma, 1996; 13(7): 343-59.
Bilgen M, Dogan B, Narayana PA. In vivo assessment of bloodspinal cord barrier permeability: serial dynamic contrast enhanced MRI of spinal cord injury. J. Magn Reson Imaging. 2002; 20(4): 337–341.
Chan CM. Inflammation: beneficial or detrimental after spinal cord injury?J.CNS drug discovery. 2008; 3(3): 189-199.
Dailey AT, Avellino AM, et al. Complement depletion reduces macrophage infiltration and activation during wallerian degeneration and axonal regeneration. J. Neurosci. 1998; 18(17): 6713–6722.
Daniel P A, Zhen Guan and Popovich P G. B cell produce pathogenic antibodies and impair recovery after spinal cord injury in mice. J. Clinical Ivestigation. 2009; 119(10): 2990-2999.
Donnelly DJ, Popovich PG. Inflammation and its role in neuroprotection, axonal regeneration and function recovery after spinal cord injury. J. Exp Neurol, 2008; 209(2): 378-88.
Chen A, Xu XM, et al. Methylprednisolone administration improves exonal regeneration into Schwann cell grafts in transected adult rat thoratic spinal cord. J. Exp neurol. 1996; 138(2): 261-76.
Fitch MT, Silver J. CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. J. Exp Neurol, 2008; 209(2): 294-301.
Frenkel D, Maron R, et al. Nasal vaccination with a proteosome-based adjuvant and glatiramer acetate clears beta-amyloid in a mouse model of Alzheimer disease. J. Clin Invest.2005; 115(9): 2423–2433.
Ghirnikar RS, Lee YL, and Eng LF. Chemokine antagonist infusion promotes axonal sparing after spinal cord contusion injury in rat. J. Neurosci. Res. 2002; 64(6): 582–589.
Gordon S. Alternative activation of macrophages. J. Nat. Rev Immunol. 2003; 3(3): 23–35.
Gris D, Marsh DR, Oatway MA and et al. Transient blockade of the CD11d/CD18 integrin reduce secondary damage after spinal cord injury,improving sensory, autonomic, and motor function. J. Neurosci. 2004; 24(15): 4043-4051.
Hamann D, Roos M, Lier RAW., Faces phases of human CD8+ T-cell development. J. Immunol Today. 1999; 20(4): 177-179.
Hsieh CS, Heimberger AB,Gold JS, et al. Differential regulation of T helper phenotype development by interleukins 4 and 10 in an alpha beta T-cell-receptor transgenic system. J. Immunology, 1992; 89(13): 6065-69.
Jasonr P, Yu Zheng, et al. Augmented Locomotor Recovery after Spinal Cord Injury in the Athymic Nude Rat. J. Neurotrauma, 2006; 23, (5): 667-673.
Jones TB, Basso DM, Sodhi A, et al. Pathological CNS autoimmune disease triggered by traumatic spinal cord injury: implications for autoimmune vaccine therapy. J. Neurosci, 2002; 22(7): 2690-700.
Keith AC, Howard EG, et al. Debate:―Is Increasing Neuroinflammation Beneficial for Neural Repair?‖J. Neuroimmune Pharmacol. 2006; 1(3): 195–211.
Kenneth M, Murphy AB, Heimberger, et al. Induction by antigen of intrathymic apoptosis of CD4+ CD8+ TCR thymocytes in vivo. J. Science, 1990; 250(4988):1720-23.
Kigerl KA, Mcgaughy VM, Popovich PG. Comparative analysis of lesion development and intraspinal inflammation in four strains of mice following spinal contusion injury. J. comp Neurol, 2006; 494(4): 578-94.
Kim HJ, Krenn V, Steinhauser G, Berek C. Plasma cell development in synovial germinal centers in patients with rheumatoid and reactive arthritis. J. Immunol. 1999; 162(5): 3053–3062.
Kipnis J, Schwartz M. Dual action of glatiramer acetate (Cop-1) in the treatment of CNS autoimmune and neuro-degenerative disorders. J. Trends Mol Med.2002; 8(7): 319–323.
Kipnis J, Yoles E, et al. T cell immunity to copolymer 1 confers neuroprotection on the damaged optic nerve: possible therapy for optic neuropathies. Proc Natl Acad Sci USA. 2000; 97(13): 7446–7451.
Knopf PM, Harling-Berg CJ, et al. Antigen-dependent intrathecal antibody synthesis in the normal rat brain: tissue entry and local retention of antigen-specific B cells. J. Immunol. 1998; 161(2): 692–701.
Krogsgaard M, Li QJ, et al. Agonist/endogenous peptide-MHC heterodimers drive T cell activation and sensitivity. J. Nature. 2005; 434(7030): 238–243..
Liu X, Xu XM, et al. Neuronal and glial apoptosis after traumatic spinal cord injury. J. Neurosci. 1997; 17(14): 5395-5406.
Lee YL, Shih K, et al. Cytokine chemokine expression in contused rat spinal cord. J. Neurochem Int. 2000; 36(4-5): 417–425.
Ling C, Sandor M, Fabry Z. In situ processing and distribution of intracerebrally injected OVA in the CNS. J. Neuroimmunol. 2003; 141(1-2): 90–98.
Ma M, Wei P, Wei T, et al. Enhanced axonal growth into a spinal cord contusion injury site in a strain of mouse (129X1/SvJ) with a diminished inflammatory response. J. Comp. Neurol. 2004; 474(4): 469–486.
Mantovani A, et al. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. J. Trends Immunol. 2002; 23(11): 549–555.
Martin P, Souza D, Martin J, et al. Wound healing in the PU.1 null mouse-tissue repair is not dependent on inflammatory cells. J. Curr Biol. 2003; 13(13): 1122–1128.
McTigue DM, Tani M, et al. Selective chemokine mRNA accumulation in the rat spinal cord after contusion injury. J. Neurosci Res. 1998; 53(3): 368–376.
Mecklenbrauker I, Kalled SL, et al. Regulation of B-cell survival by BAFF-dependent PKC deltamediated nuclear signalling. J. Nature. 2004; 431(7007): 456–461.
Moalem G, Monsonego A, et al. Differential T cell response in central and peripheral nerve injury: connection with immune privilege. J. FASEB. 1999a; 13(10): 1207–1217.
Moalem G, Leibowitz-Amit R, Schwartz M, et al. Autoimmune T cells protect neurons from secondary degeneration after central nervous system axotomy. J. Nat Med. 1999b; 5(1): 49–55.
Moriya T, Hassan AZ, et al. Dynamics of extracellular calcium activity following contusion of the rat spinal cord. 1994, 11(3): 255-263.
Mosser DM. The many faces of macrophage activation. J. Leukoc. Biol. 2003; 73(2): 209–212.
Park E, Velumian, AA, and Fehlings MG, et al. The role of excitotoxicity in secondary mechanisms of spinal cord injury: a review with an emphasis on the im plications for white matter degeneration. J. Neurotrauma., 2004; 21(6): 754–74.
Plemel JR, Duncan G, et al .A graded forceps crush spinal cord injury model in mice. J. neurotrauma, 2008; 25(4): 350-70.
Popovich PG, Guan Z, et al. The neuropathological and behavioral consequences of intraspinal microglial/macrophage activation. J. Neuropathol. Exp. Neurol. 2002; 61(7): 623–633.
Profyris C, Cheema SS, et al. Degenerative and regenerative mechanisms governing spinal cord injury. J. Neurobiol. Dis. 2004; 15(3): 415–436.
Rolls A, Shechter R, Schwartz M. The bright side of the glial scar in CNS repair. J .Nat Rev Neurosci, 2009; 10(3): 235-41.
Samira Saadoun, B. Anthony Bell, et al. Greatly improved neurological outcome after spinal cord compression injury in AQP4-deficient mice. J. Brain, 2008; 131(pt4): 1087-1098.
Schmitt AB, Buss A, et al. Major histocompatibility complex class II expression by activated microglia caudal to lesions of descending tracts in the human spinal cord is not associated with a T cell response. J. Acta Neuropathol (Berl) 2000; 100(5): 528–536.
Schnell L, Schneider R, et al. Lymphocyte recruitment following spinal cord injury in mice is altered by prior viral exposure. J. Neurosci. 1997; 9(5): 1000–1007.
Schnell L, Fearn S, et al. Acute inflammatory responses to mechanical lesions in the CNS: differences between brain and spinal cord. J. Eur J Neurosci. 1999; 11(10): 3648–3658.
Schroder AE, Greiner A, et al. Differentiation of B cells in the nonlymphoid tissue of the synovial membrane of patients with rheumatoid arthritis. J. Proc Natl Acad Sci U S A. 1996; 93(1): 221–225.
Schwab JM, Brechtel K, et al. Experimental strategies to promote spinal cord regeneration-an integrative perspective. J. Prog Neurobiol. 2006; 78(2): 91-116.
Schwartz M. Beneficial autoimmune T cells and posttraumatic neuroprotection. J. Ann N Y Acad Sci 2000a; 917: 341–347.
Schwartz M. Autoimmune involvement in CNS trauma is beneficial if well controlled. J. Prog Brain Res. 2000b; 128: 259–263.
Schwartz M, Kipnis J. Therapeutic T cell-based vaccination for neurodegenerative disorders: the role of CD4+CD25+ regulatory T cells. J. Ann N Y Acad Sci. 2005; 1051:701–708.
Schwartz M, Yoles E. Immune-based therapy for spinal cord repair: autologous macrophages and beyond. J. Neurotrauma, 2006; 23(3-4): 360-70.
Springer JE, Leon A, et al. Activition of the caspase-3 apoptosis cascade in traumatic spinal cord injury. J. Nat Med.1999; 5(8): 943-946.
Serafini B, Rosicarelli B, et al. Detection of ectopic B-cell follicles with germinal centers in the meninges of patients with secondary progressive multiple sclerosis. J. Brain Pathol. 2004; 14(2): 164–174.
Sroga JM, Jones TB. Rats and mice exhibit distinct inflammatory reactions after spinalcord injury. J. Comp Neurol. 2003; 462(2): 223–240.
Stefanova I, Dorfman JR, Germain RN. Self-recognition promotes the foreign antigen sensitivity of naive T lymphocytes. J. Nature. 2002; 420(6914): 429–434.
Taoka Y, Naruo M, et al. Superoxide radicals play important role in the pathogenesis of spinal cord injury. J. Paraplegia. 1995; 33(8): 450-453.
Tornqvist E, Liu L, et al. Complement and clusterin in the injured nervous system. J. Neurobiol Aging. 1996; 17(5): 695–705.
Whetstone WD, Hsu JY, et al. Blood-spinal cord barrier after spinal cord injury: relation to revascularization and wound healing. J. Neurosci Res 2003; 74(2): 227–239.
Young W. Role of calcium in central nervous system injuries. J. Neurotrauma. 1992; 8: S9-25.
Anderson AJ, Robert S, et al. Activation of complement pathways after contusion-induced spinal cord injury. J. Neurotrauma. 2004; 21(12): 1831–1846.
Ankeny DP, Popovich PG, et al. Mechanisms and implications of adaptive immune responses after traumatic spinal cord injury. J. Neuroscience, 2009; 158(3): 1112–21.
Arumugam T, Woodruff TM, et al. Neuroprotection in stroke by complement inhibition and immunoglobulin therapy. J. Neuroscience. 2009; 158(3): 1074–1089.
Bakalash S, Shlomo GB, et al. T-cell based vaccination for morphological and functional neuroprotection in a rat model of chronically elevated intraocular pressure. J. Mol Med. 2005; 83(11): 904–916.
Barnum SR. Inhibition of complement as a therapeutic approach in inflammatory central nervous system (CNS) disease. J. Mol Med. 1999; 5(9): 569–582.
Basso DM, Beattie MS, Bresnahan JC. MASCIS evaluation of open field locomotor scores: Effects of Experience and Teamwork on Reliability. J. Neurotroma, 1996; 13(7): 343-59.
Bilgen M, Dogan B, Narayana PA. In vivo assessment of bloodspinal cord barrier permeability: serial dynamic contrast enhanced MRI of spinal cord injury. J. Magn Reson Imaging. 2002; 20(4): 337–341.
Chan CM. Inflammation: beneficial or detrimental after spinal cord injury?J.CNS drug discovery. 2008; 3(3): 189-199.
Dailey AT, Avellino AM, et al. Complement depletion reduces macrophage infiltration and activation during wallerian degeneration and axonal regeneration. J. Neurosci. 1998; 18(17): 6713–6722.
Daniel P A, Zhen Guan and Popovich P G. B cell produce pathogenic antibodies and impair recovery after spinal cord injury in mice. J. Clinical Ivestigation. 2009; 119(10): 2990-2999.
Donnelly DJ, Popovich PG. Inflammation and its role in neuroprotection, axonal regeneration and function recovery after spinal cord injury. J. Exp Neurol, 2008; 209(2): 378-88.
Chen A, Xu XM, et al. Methylprednisolone administration improves exonal regeneration into Schwann cell grafts in transected adult rat thoratic spinal cord. J. Exp neurol. 1996; 138(2): 261-76.
Fitch MT, Silver J. CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. J. Exp Neurol, 2008; 209(2): 294-301.
Frenkel D, Maron R, et al. Nasal vaccination with a proteosome-based adjuvant and glatiramer acetate clears beta-amyloid in a mouse model of Alzheimer disease. J. Clin Invest.2005; 115(9): 2423–2433.
Ghirnikar RS, Lee YL, and Eng LF. Chemokine antagonist infusion promotes axonal sparing after spinal cord contusion injury in rat. J. Neurosci. Res. 2002; 64(6): 582–589.
Gordon S. Alternative activation of macrophages. J. Nat. Rev Immunol. 2003; 3(3): 23–35.
Gris D, Marsh DR, Oatway MA and et al. Transient blockade of the CD11d/CD18 integrin reduce secondary damage after spinal cord injury,improving sensory, autonomic, and motor function. J. Neurosci. 2004; 24(15): 4043-4051.
Hamann D, Roos M, Lier RAW., Faces phases of human CD8+ T-cell development. J. Immunol Today. 1999; 20(4): 177-179.
Hsieh CS, Heimberger AB,Gold JS, et al. Differential regulation of T helper phenotype development by interleukins 4 and 10 in an alpha beta T-cell-receptor transgenic system. J. Immunology, 1992; 89(13): 6065-69.
Jasonr P, Yu Zheng, et al. Augmented Locomotor Recovery after Spinal Cord Injury in the Athymic Nude Rat. J. Neurotrauma, 2006; 23, (5): 667-673.
Jones TB, Basso DM, Sodhi A, et al. Pathological CNS autoimmune disease triggered by traumatic spinal cord injury: implications for autoimmune vaccine therapy. J. Neurosci, 2002; 22(7): 2690-700.
Keith AC, Howard EG, et al. Debate:―Is Increasing Neuroinflammation Beneficial for Neural Repair?‖J. Neuroimmune Pharmacol. 2006; 1(3): 195–211.
Kenneth M, Murphy AB, Heimberger, et al. Induction by antigen of intrathymic apoptosis of CD4+ CD8+ TCR thymocytes in vivo. J. Science, 1990; 250(4988):1720-23.
Kigerl KA, Mcgaughy VM, Popovich PG. Comparative analysis of lesion development and intraspinal inflammation in four strains of mice following spinal contusion injury. J. comp Neurol, 2006; 494(4): 578-94.
Kim HJ, Krenn V, Steinhauser G, Berek C. Plasma cell development in synovial germinal centers in patients with rheumatoid and reactive arthritis. J. Immunol. 1999; 162(5): 3053–3062.
Kipnis J, Schwartz M. Dual action of glatiramer acetate (Cop-1) in the treatment of CNS autoimmune and neuro-degenerative disorders. J. Trends Mol Med.2002; 8(7): 319–323.
Kipnis J, Yoles E, et al. T cell immunity to copolymer 1 confers neuroprotection on the damaged optic nerve: possible therapy for optic neuropathies. Proc Natl Acad Sci USA. 2000; 97(13): 7446–7451.
Knopf PM, Harling-Berg CJ, et al. Antigen-dependent intrathecal antibody synthesis in the normal rat brain: tissue entry and local retention of antigen-specific B cells. J. Immunol. 1998; 161(2): 692–701.
Krogsgaard M, Li QJ, et al. Agonist/endogenous peptide-MHC heterodimers drive T cell activation and sensitivity. J. Nature. 2005; 434(7030): 238–243..
Liu X, Xu XM, et al. Neuronal and glial apoptosis after traumatic spinal cord injury. J. Neurosci. 1997; 17(14): 5395-5406.
Lee YL, Shih K, et al. Cytokine chemokine expression in contused rat spinal cord. J. Neurochem Int. 2000; 36(4-5): 417–425.
Ling C, Sandor M, Fabry Z. In situ processing and distribution of intracerebrally injected OVA in the CNS. J. Neuroimmunol. 2003; 141(1-2): 90–98.
Ma M, Wei P, Wei T, et al. Enhanced axonal growth into a spinal cord contusion injury site in a strain of mouse (129X1/SvJ) with a diminished inflammatory response. J. Comp. Neurol. 2004; 474(4): 469–486.
Mantovani A, et al. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. J. Trends Immunol. 2002; 23(11): 549–555.
Martin P, Souza D, Martin J, et al. Wound healing in the PU.1 null mouse-tissue repair is not dependent on inflammatory cells. J. Curr Biol. 2003; 13(13): 1122–1128.
McTigue DM, Tani M, et al. Selective chemokine mRNA accumulation in the rat spinal cord after contusion injury. J. Neurosci Res. 1998; 53(3): 368–376.
Mecklenbrauker I, Kalled SL, et al. Regulation of B-cell survival by BAFF-dependent PKC deltamediated nuclear signalling. J. Nature. 2004; 431(7007): 456–461.
Moalem G, Monsonego A, et al. Differential T cell response in central and peripheral nerve injury: connection with immune privilege. J. FASEB. 1999a; 13(10): 1207–1217.
Moalem G, Leibowitz-Amit R, Schwartz M, et al. Autoimmune T cells protect neurons from secondary degeneration after central nervous system axotomy. J. Nat Med. 1999b; 5(1): 49–55.
Moriya T, Hassan AZ, et al. Dynamics of extracellular calcium activity following contusion of the rat spinal cord. 1994, 11(3): 255-263.
Mosser DM. The many faces of macrophage activation. J. Leukoc. Biol. 2003; 73(2): 209–212.
Park E, Velumian, AA, and Fehlings MG, et al. The role of excitotoxicity in secondary mechanisms of spinal cord injury: a review with an emphasis on the im plications for white matter degeneration. J. Neurotrauma., 2004; 21(6): 754–74.
Plemel JR, Duncan G, et al .A graded forceps crush spinal cord injury model in mice. J. neurotrauma, 2008; 25(4): 350-70.
Popovich PG, Guan Z, et al. The neuropathological and behavioral consequences of intraspinal microglial/macrophage activation. J. Neuropathol. Exp. Neurol. 2002; 61(7): 623–633.
Profyris C, Cheema SS, et al. Degenerative and regenerative mechanisms governing spinal cord injury. J. Neurobiol. Dis. 2004; 15(3): 415–436.
Rolls A, Shechter R, Schwartz M. The bright side of the glial scar in CNS repair. J .Nat Rev Neurosci, 2009; 10(3): 235-41.
Samira Saadoun, B. Anthony Bell, et al. Greatly improved neurological outcome after spinal cord compression injury in AQP4-deficient mice. J. Brain, 2008; 131(pt4): 1087-1098.
Schmitt AB, Buss A, et al. Major histocompatibility complex class II expression by activated microglia caudal to lesions of descending tracts in the human spinal cord is not associated with a T cell response. J. Acta Neuropathol (Berl) 2000; 100(5): 528–536.
Schnell L, Schneider R, et al. Lymphocyte recruitment following spinal cord injury in mice is altered by prior viral exposure. J. Neurosci. 1997; 9(5): 1000–1007.
Schnell L, Fearn S, et al. Acute inflammatory responses to mechanical lesions in the CNS: differences between brain and spinal cord. J. Eur J Neurosci. 1999; 11(10): 3648–3658.
Schroder AE, Greiner A, et al. Differentiation of B cells in the nonlymphoid tissue of the synovial membrane of patients with rheumatoid arthritis. J. Proc Natl Acad Sci U S A. 1996; 93(1): 221–225.
Schwab JM, Brechtel K, et al. Experimental strategies to promote spinal cord regeneration-an integrative perspective. J. Prog Neurobiol. 2006; 78(2): 91-116.
Schwartz M. Beneficial autoimmune T cells and posttraumatic neuroprotection. J. Ann N Y Acad Sci 2000a; 917: 341–347.
Schwartz M. Autoimmune involvement in CNS trauma is beneficial if well controlled. J. Prog Brain Res. 2000b; 128: 259–263.
Schwartz M, Kipnis J. Therapeutic T cell-based vaccination for neurodegenerative disorders: the role of CD4+CD25+ regulatory T cells. J. Ann N Y Acad Sci. 2005; 1051:701–708.
Schwartz M, Yoles E. Immune-based therapy for spinal cord repair: autologous macrophages and beyond. J. Neurotrauma, 2006; 23(3-4): 360-70.
Springer JE, Leon A, et al. Activition of the caspase-3 apoptosis cascade in traumatic spinal cord injury. J. Nat Med.1999; 5(8): 943-946.
Serafini B, Rosicarelli B, et al. Detection of ectopic B-cell follicles with germinal centers in the meninges of patients with secondary progressive multiple sclerosis. J. Brain Pathol. 2004; 14(2): 164–174.
Sroga JM, Jones TB. Rats and mice exhibit distinct inflammatory reactions after spinalcord injury. J. Comp Neurol. 2003; 462(2): 223–240.
Stefanova I, Dorfman JR, Germain RN. Self-recognition promotes the foreign antigen sensitivity of naive T lymphocytes. J. Nature. 2002; 420(6914): 429–434.
Taoka Y, Naruo M, et al. Superoxide radicals play important role in the pathogenesis of spinal cord injury. J. Paraplegia. 1995; 33(8): 450-453.
Tornqvist E, Liu L, et al. Complement and clusterin in the injured nervous system. J. Neurobiol Aging. 1996; 17(5): 695–705.
Whetstone WD, Hsu JY, et al. Blood-spinal cord barrier after spinal cord injury: relation to revascularization and wound healing. J. Neurosci Res 2003; 74(2): 227–239.
Young W. Role of calcium in central nervous system injuries. J. Neurotrauma. 1992; 8: S9-25.