Th1/Th17在小鼠异种周围神经移植急性排斥反应中作用的研究
摘要
周围神经损伤是临床的常见病、多发病，尤以神经缺损多见，如果不能及时修复缺损，恢复神经的连续性及完整性，会严重影响患者的感觉及运动功能。在临床大量的病例中，长段的周围神经缺损的神经断端不能直接端端缝合，通常需要自体神经移植或同种异体神经移植来桥接神经缺损。虽然自体神经移植被认为是修复神经缺损的最有效方法，但是却存在着一定的缺陷，如供体来源有限、供区的副损伤、瘢痕形成及神经瘤的形成等。同种异体神经移植也因其来源有限且存在一定的伦理问题，使其临床应用受到限制。为寻找修复周围神经缺损的有效方法，异种周围神经移植越来越受到人们的重视。
与心脏、肾脏等大型实体器官移植不同，异种神经移植体作为受体自身轴突生长的暂时性引导通路，仅在移植后早期发挥作用。移植后，如果受体自身的神经已经完成再生，达到支配效应器官的目的，则移植体存在与否已无意义。因此，异种神经移植早期排斥反应对手术成功与否具有重要意义。目前研究认为，1型辅助性T淋巴细胞相关的细胞因子是移植排斥反应的主要细胞因子，可通过促进移植体抗原特异性的细胞毒性T细胞（CTL）和迟发型超敏性T细胞（TDTH）启动排斥反应，而Th2细胞则可抑制Th1细胞的分化及功能，抑制排斥反应。Th1/Th2轴作为移植排斥机制的核心已被大多数人接受。近年研究已经发现，Th17及其细胞因子IL-17在肾脏、心脏、肺及肝脏等器官的移植中也发挥了重要的作用。
在异种周围神经移植中，Th17和Th1分别是通过何种途径产生移植排斥反应的？它们中到底哪一种在起主导作用呢？它们之间是否具有协同或者拮抗作用？通过中和抗体的特异性阻断，能否帮助移植体逃脱受体的免疫监控，从而避免排斥反应的发生呢？本研究应用大鼠到小鼠的异种周围神经移植动物模型210例，得到以下研究结果:
1.异种周围神经移植后早期，神经移植物周围的单核细胞浸润显著增加，且IFN-γ+、IL-17+细胞浸润增加；血清中的IFN-γ、IL-17含量明显升高；脾脏淋巴细胞胞浆内的IFN-γ、IL-17升高；而IL-4在血清、脾脏淋巴细胞及神经移植物周围的变化均无统计学意义。
2.在应用IFN-γ中和抗体之后，虽然脾脏中的IFN-γ、IL-17表达较未应用中和抗体的异种移植无明显差异，但神经移植物周围的单核细胞、IFN-γ+、IL-17+细胞浸润均明显减少。
3.应用IL-17中和抗体之后，血清、脾脏及神经移植物周围的IFN-γ、IL-17均减少，且单核细胞浸润也减少。
4.联合应用IFN-γ、IL-17中和抗体的结果与单独应用IL-17中和抗体的结果相似。
5.IFN-γ、IL-17在异种周围神经移植早期的急性排斥反应中发挥重要作用，且Th17可不依赖于Th1而单独发挥作用；阻断IFN-γ和IL-17的效应后，可以有效地抑制异种周围神经移植早期急性排斥反应。
本研究初步证实了Th1/Th17细胞介导了异种周围神经移植早期急性排斥反应，且Th17可不依赖于Th1而单独发挥作用；应用抗IFN-γ、IL-17中和抗体，可有效抑制异种周围神经移植早期急性排斥反应，从而为临床有效干预提供了可行性依据。虽然可以确定Th1及Th17在异种周围神经移植排斥反应早期发挥重要作用，但是两者之间的调节机制，以及与CD8+T淋巴细胞之间的关系、与固有免疫细胞之间是否存在其它的调节作用，尚有待进一步研究。
Peripheral nerve injuries are very commonly seen in the clinic. If they cannot be immediately repaired to recover continuity and integrity, they often result in motor and sensory deficits for patients. In many instances these injuries result in long nerve segment loss preventing the nerve ends from being directly sutured due to increased tension. In these cases grafts such as autografts and allografts must be used to bridge the long nerve gaps. Although autografts are recognized as the gold standard for nerve grafting, they have several limitations including limited donor source, functional loss in the distribution of the donor nerve, scarring, and painful neuroma formation. Allografts also have defects in limitations of source. In order to find a means of reconstructing injured nerves, peripheral nerve xenotransplantation has become an increasingly promising alternative.
Compared with heart, renal and other organs transplantation, peripheral nerve xenografts are just as the tube for nerve regeneration and play the role at the primary period. If the injuried nerve have regenerated and controlled the muscle, whether the xenografts exist will not be important for recipients. The cytokines secreted by thl cells are recognized as the major factors, and can promote the transplant rejection induced by the specific CTL and TDTH. Th2cells can inhibit the function of Th1. Th1/Th2paradigm is the center of the rejection mechanism and has been accepted by researchers. Recent studies demonstrate that Thl7and IL-17take part in the rejection of renal, heart, lung and liver transplantation.
During peripheral nerve xenotransplantation, what is the regulatory mechanism between Thl and Thl7; what kind of cells plays the major role; whether the rejection can be prevented by blocking the role of these two kinds of cells? I performed the peripheral nerve xenotransplantation from rats to mouse, and get the conclusion:
1Compared with peripheral nerve autotransplantation, mononuclear cells invasion increased during peripheral never xenotransplantation acute rejection; IFN-γ+、IL-17+cells invasion also increased around the surrounding tissues of xenografts; the levels of IFN-γ、IL-17increased in serum; the expression of IFN-γ、IL-17in lymphocytes from spleen increased. The changes of IL-4had no significant differences.
2After using IFN-y neutralizing antibodies, although there is no significant difference of IFN-γ、IL-17expression in serum and lymphocytes from spleen compared to the xenografts recipients without injecting neutralizing antibodies, IFN-γ+、IL-17+cells invasion decreased around the surrounding tissues of xenografts.
3After using IL-17neutralizing antibodies,IFN-γ、IL-17decreased in serum, spleen and surrounding tissues of xenografts.
4The results in xenografts recipients of IFN-γ and IL-17neutralizing antibodies mixed used were similar as the recipients with IL-17neutralizing antibodies single used.
5IFN-γ and IL-17play the crucial role during the acute rejection of peripheral nerve xenotransplantation; Thl7can play the role respectively, and are not dependent Thl; the rejection of peripheral nerve xenotransplantation by blocking the function of IFN-γ and IL-17.
This study is the primary research about the role of Thl/Thl7during acute rejection of peripheral nerve xenotranspantation, and about the mechanism of rejection after using IFN-γ and IL-17neutralizing antibodies. The conclusion can provide the data and method for preventing the rejection and clinical treatment. Although Thl and Thl7play the role during the rejection, the regulatory mechanism between these two kinds of cells and the relationship between innate and adaptive immune cells need to be further studied. I hope do my best for clinical use of peripheral nerve xeontransplantation.
与心脏、肾脏等大型实体器官移植不同，异种神经移植体作为受体自身轴突生长的暂时性引导通路，仅在移植后早期发挥作用。移植后，如果受体自身的神经已经完成再生，达到支配效应器官的目的，则移植体存在与否已无意义。因此，异种神经移植早期排斥反应对手术成功与否具有重要意义。目前研究认为，1型辅助性T淋巴细胞相关的细胞因子是移植排斥反应的主要细胞因子，可通过促进移植体抗原特异性的细胞毒性T细胞（CTL）和迟发型超敏性T细胞（TDTH）启动排斥反应，而Th2细胞则可抑制Th1细胞的分化及功能，抑制排斥反应。Th1/Th2轴作为移植排斥机制的核心已被大多数人接受。近年研究已经发现，Th17及其细胞因子IL-17在肾脏、心脏、肺及肝脏等器官的移植中也发挥了重要的作用。
在异种周围神经移植中，Th17和Th1分别是通过何种途径产生移植排斥反应的？它们中到底哪一种在起主导作用呢？它们之间是否具有协同或者拮抗作用？通过中和抗体的特异性阻断，能否帮助移植体逃脱受体的免疫监控，从而避免排斥反应的发生呢？本研究应用大鼠到小鼠的异种周围神经移植动物模型210例，得到以下研究结果:
1.异种周围神经移植后早期，神经移植物周围的单核细胞浸润显著增加，且IFN-γ+、IL-17+细胞浸润增加；血清中的IFN-γ、IL-17含量明显升高；脾脏淋巴细胞胞浆内的IFN-γ、IL-17升高；而IL-4在血清、脾脏淋巴细胞及神经移植物周围的变化均无统计学意义。
2.在应用IFN-γ中和抗体之后，虽然脾脏中的IFN-γ、IL-17表达较未应用中和抗体的异种移植无明显差异，但神经移植物周围的单核细胞、IFN-γ+、IL-17+细胞浸润均明显减少。
3.应用IL-17中和抗体之后，血清、脾脏及神经移植物周围的IFN-γ、IL-17均减少，且单核细胞浸润也减少。
4.联合应用IFN-γ、IL-17中和抗体的结果与单独应用IL-17中和抗体的结果相似。
5.IFN-γ、IL-17在异种周围神经移植早期的急性排斥反应中发挥重要作用，且Th17可不依赖于Th1而单独发挥作用；阻断IFN-γ和IL-17的效应后，可以有效地抑制异种周围神经移植早期急性排斥反应。
本研究初步证实了Th1/Th17细胞介导了异种周围神经移植早期急性排斥反应，且Th17可不依赖于Th1而单独发挥作用；应用抗IFN-γ、IL-17中和抗体，可有效抑制异种周围神经移植早期急性排斥反应，从而为临床有效干预提供了可行性依据。虽然可以确定Th1及Th17在异种周围神经移植排斥反应早期发挥重要作用，但是两者之间的调节机制，以及与CD8+T淋巴细胞之间的关系、与固有免疫细胞之间是否存在其它的调节作用，尚有待进一步研究。
Peripheral nerve injuries are very commonly seen in the clinic. If they cannot be immediately repaired to recover continuity and integrity, they often result in motor and sensory deficits for patients. In many instances these injuries result in long nerve segment loss preventing the nerve ends from being directly sutured due to increased tension. In these cases grafts such as autografts and allografts must be used to bridge the long nerve gaps. Although autografts are recognized as the gold standard for nerve grafting, they have several limitations including limited donor source, functional loss in the distribution of the donor nerve, scarring, and painful neuroma formation. Allografts also have defects in limitations of source. In order to find a means of reconstructing injured nerves, peripheral nerve xenotransplantation has become an increasingly promising alternative.
Compared with heart, renal and other organs transplantation, peripheral nerve xenografts are just as the tube for nerve regeneration and play the role at the primary period. If the injuried nerve have regenerated and controlled the muscle, whether the xenografts exist will not be important for recipients. The cytokines secreted by thl cells are recognized as the major factors, and can promote the transplant rejection induced by the specific CTL and TDTH. Th2cells can inhibit the function of Th1. Th1/Th2paradigm is the center of the rejection mechanism and has been accepted by researchers. Recent studies demonstrate that Thl7and IL-17take part in the rejection of renal, heart, lung and liver transplantation.
During peripheral nerve xenotransplantation, what is the regulatory mechanism between Thl and Thl7; what kind of cells plays the major role; whether the rejection can be prevented by blocking the role of these two kinds of cells? I performed the peripheral nerve xenotransplantation from rats to mouse, and get the conclusion:
1Compared with peripheral nerve autotransplantation, mononuclear cells invasion increased during peripheral never xenotransplantation acute rejection; IFN-γ+、IL-17+cells invasion also increased around the surrounding tissues of xenografts; the levels of IFN-γ、IL-17increased in serum; the expression of IFN-γ、IL-17in lymphocytes from spleen increased. The changes of IL-4had no significant differences.
2After using IFN-y neutralizing antibodies, although there is no significant difference of IFN-γ、IL-17expression in serum and lymphocytes from spleen compared to the xenografts recipients without injecting neutralizing antibodies, IFN-γ+、IL-17+cells invasion decreased around the surrounding tissues of xenografts.
3After using IL-17neutralizing antibodies,IFN-γ、IL-17decreased in serum, spleen and surrounding tissues of xenografts.
4The results in xenografts recipients of IFN-γ and IL-17neutralizing antibodies mixed used were similar as the recipients with IL-17neutralizing antibodies single used.
5IFN-γ and IL-17play the crucial role during the acute rejection of peripheral nerve xenotransplantation; Thl7can play the role respectively, and are not dependent Thl; the rejection of peripheral nerve xenotransplantation by blocking the function of IFN-γ and IL-17.
This study is the primary research about the role of Thl/Thl7during acute rejection of peripheral nerve xenotranspantation, and about the mechanism of rejection after using IFN-γ and IL-17neutralizing antibodies. The conclusion can provide the data and method for preventing the rejection and clinical treatment. Although Thl and Thl7play the role during the rejection, the regulatory mechanism between these two kinds of cells and the relationship between innate and adaptive immune cells need to be further studied. I hope do my best for clinical use of peripheral nerve xeontransplantation.
引文
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