腎移植受者血清多反應(yīng)性抗體與HLA抗體的相關(guān)性及對(duì)移植腎長(zhǎng)期存活的影響
發(fā)布時(shí)間：2018-07-28 12:46
【摘要】：抗體介導(dǎo)的移植物排異反應(yīng)是導(dǎo)致移植物失功的主要原因，也是影響移植腎長(zhǎng)期存活的關(guān)鍵性因素。目前廣泛認(rèn)為，器官移植后移植物發(fā)生抗體介導(dǎo)排異反應(yīng)及移植物失功主要與受者DSA的產(chǎn)生密切相關(guān)，而NDSA及非HLA抗體對(duì)移植物存活的影響尚未得到足夠重視。有文獻(xiàn)報(bào)道，器官移植受者血清中未產(chǎn)生DSA的情況下，仍然存在針對(duì)移植物的抗體介導(dǎo)的排異反應(yīng)[4,5]。另有研究報(bào)道，器官移植患者體內(nèi)諸如血管緊張素受體、抗波形蛋白抗體、抗過(guò)氧化脂類抗體等諸多自身抗體與移植物的排異反應(yīng)密切相關(guān)并影響移植物長(zhǎng)期存活[7-10]。綜上所述，除了供體特異性抗體之外，非供體特異性HLA抗體及非HLA抗體也是影響移植物存活的重要因素。 器官移植致敏后產(chǎn)生的抗體主要為特異性抗供者HLA的抗體，然而在研究中發(fā)現(xiàn)，腎移植受者血清中亦可檢測(cè)出NDSA的出現(xiàn)[6,11-13]，在無(wú)特異性免疫致敏的前提下，NDSA產(chǎn)生的原因尚不完全明確。目前普遍接受的解釋為交叉反應(yīng)原理[14-17]，即由于部分HLA之間存在共用的抗原決定簇，故腎移植術(shù)后患者血清中產(chǎn)生特異性抗供者HLA抗體的同時(shí)，也可以與供者表達(dá)的HLA相似的抗原發(fā)生反應(yīng)，從而表現(xiàn)為NDSA的出現(xiàn)。然而有研究發(fā)現(xiàn)，腎移植患者在沒(méi)有產(chǎn)生DSA的情況下也可檢測(cè)出NDSA，腎移植術(shù)后NDSA出現(xiàn)的平均中位時(shí)間甚至早于DSA[14]。此種現(xiàn)象顯然不能用現(xiàn)有的交叉反應(yīng)理論來(lái)解釋。 自然抗體（Natural antibodies）是一類未經(jīng)免疫致敏而廣泛存在于人類血清中的抗體[18]。自然抗體由于能夠與抗原決定簇毫不相關(guān)的多種抗原均發(fā)生反應(yīng)，故表現(xiàn)出多反應(yīng)性的特點(diǎn)，從而在人類先天免疫中發(fā)揮著重要的作用。自然抗體的重要特性是能夠與凋亡細(xì)胞結(jié)合[19-22]。 長(zhǎng)期以來(lái)，對(duì)多反應(yīng)性抗體的研究主要集中于人類自身免疫性疾病領(lǐng)域，而針對(duì)多反應(yīng)性抗體在器官移植領(lǐng)域的研究卻罕有報(bào)道。鑒于多反應(yīng)性抗體的特性，在本研究中，我們分別從單克隆B細(xì)胞、抗體水平以及腎移植患者血清學(xué)水平探尋多反應(yīng)性抗體與HLA抗體產(chǎn)生的相關(guān)性。同時(shí)研究腎移植受者血清內(nèi)預(yù)存的多反應(yīng)性抗體與抗體介導(dǎo)移植腎排異反應(yīng)的相關(guān)性及對(duì)移植腎長(zhǎng)期存活的影響。結(jié)果如下： 1、多反應(yīng)性抗體與HLA抗體產(chǎn)生的相關(guān)性。 本研究中，我們將B淋巴細(xì)胞從腎移植受者外周血及手術(shù)切除的移植腎中分選出來(lái)并在體外永生化，成功培育出單克隆B淋巴細(xì)胞克隆。通過(guò)檢測(cè)發(fā)現(xiàn)所有B淋巴細(xì)胞克隆均分泌抗體，部分單克隆抗體可以與多種抗原諸如LPS、dsDNA、胰島素、細(xì)胞裂解產(chǎn)物等發(fā)生抗原抗體反應(yīng)，從而揭示此類單克隆抗體具有多反應(yīng)性的特點(diǎn)，有別于“一把鑰匙開一把鎖”的特異性抗原抗體反應(yīng)。此外，我們通過(guò)研究發(fā)現(xiàn)，，多反應(yīng)性抗體能夠與凋亡細(xì)胞結(jié)合。值得關(guān)注的是，我們發(fā)現(xiàn)部分多反應(yīng)性的單克隆抗體能夠與多種HLA抗原發(fā)生反應(yīng)，從而在單克隆抗體水平發(fā)現(xiàn)多反應(yīng)性抗體與HLA抗體產(chǎn)生的相關(guān)性。當(dāng)將研究擴(kuò)展至患者血清學(xué)水平，我們發(fā)現(xiàn)，與HLA抗體陰性的患者相比，HLA抗體陽(yáng)性的患者血清內(nèi)多反應(yīng)性抗體活性顯著升高（p=0.009，p0.001，p0.001）。應(yīng)用Spearman相關(guān)性分析發(fā)現(xiàn)300例腎移植患者血清中多反應(yīng)性抗體活性與HLA抗體活性之間存在顯著相關(guān)性（p=0.004，p0.001，p=0.019）。在HLAI類抗體陽(yáng)性的患者中，與血清多反應(yīng)性抗體活性低的患者相比，血清多反應(yīng)性抗體活性高的患者能夠與更多的HLAI類抗原發(fā)生反應(yīng)（p=0.030），因而從血清學(xué)水平揭示了多反應(yīng)性抗體與HLA抗體的相關(guān)性。 2、腎移植受者術(shù)前血清多反應(yīng)性抗體與移植腎抗體介導(dǎo)排異反應(yīng)及移植腎長(zhǎng)期存活的相關(guān)性。 我們通過(guò)對(duì)300例腎移植患者血清標(biāo)本研究發(fā)現(xiàn)，腎移植術(shù)前患者血清多反應(yīng)性抗體IgG的活性較正常人群顯著性升高（p=0.011），而這種差異并非由于腎移植患者及正常人群血清中IgG抗體濃度的不同所致。在此項(xiàng)回顧性研究中，300例患者隨訪時(shí)間為81.2±35.3個(gè)月。46例患者在觀察期內(nèi)出現(xiàn)移植腎失功，我們通過(guò)研究發(fā)現(xiàn)移植腎失功的患者術(shù)前血清多反應(yīng)性IgG活性顯著高于移植腎功能正常的患者（p0.001）。而且在46例移植腎失功的患者中，因抗體介導(dǎo)排異反應(yīng)導(dǎo)致移植腎失功的患者血清多反應(yīng)性IgG活性顯著高于因其他原因?qū)е乱浦材I失功的患者（p=0.033)。Kaplan-Meier生存曲線顯示，移植前血清中多反應(yīng)性抗體活性高的患者移植腎存活率顯著降低（p0.001），而多反應(yīng)性IgG獨(dú)立于HLA抗體之外對(duì)移植腎存活產(chǎn)生影響。進(jìn)一步行Cox比例風(fēng)險(xiǎn)模型分析顯示，血清多反應(yīng)性抗體IgG活性是移植腎失功的獨(dú)立危險(xiǎn)因素（Hazard ratio=2.271，p0.001）。 本研究發(fā)現(xiàn)，腎移植患者血清多反應(yīng)性IgG抗體對(duì)凋亡細(xì)胞的反應(yīng)主要由IgG1亞型及IgG3亞型介導(dǎo)。多反應(yīng)性抗體與凋亡細(xì)胞反應(yīng)后可激活補(bǔ)體途徑，致使C4d沉積于靶細(xì)胞表面。進(jìn)一步分析顯示，IgG1及IgG3亞型對(duì)凋亡細(xì)胞的反應(yīng)性與C4d在靶細(xì)胞表面沉積強(qiáng)度呈顯著相關(guān)性（p0.001，p=0.005）。 本研究分別從單克隆抗體水平及腎移植受者血清學(xué)水平研究發(fā)現(xiàn)多反應(yīng)性抗體與HLA抗體的產(chǎn)生具有顯著相關(guān)性，在交叉反應(yīng)理論之外，為器官移植患者HLA抗體尤其是NDSA的產(chǎn)生原因提供新的依據(jù)；并發(fā)現(xiàn)多反應(yīng)性抗體IgG與腎移植術(shù)后抗體介導(dǎo)的排異反應(yīng)顯著相關(guān)，并成為移植物失功的獨(dú)立危險(xiǎn)因素。本研究為腎移植患者術(shù)前風(fēng)險(xiǎn)評(píng)估提供了新的依據(jù)。在器官移植術(shù)前除常規(guī)檢測(cè)HLA抗體之外，對(duì)患者血清多反應(yīng)性抗體的監(jiān)測(cè)有利于提高器官移植后移植物失功風(fēng)險(xiǎn)評(píng)估的準(zhǔn)確性，并為器官移植術(shù)前高風(fēng)險(xiǎn)患者的去抗體治療提供了新的理論及實(shí)驗(yàn)依據(jù)。
[Abstract]:Antibody mediated graft rejection is the main cause of graft dysfunction and is also a key factor affecting the long-term survival of the transplanted kidney. It is widely believed that antibody mediated rejection and graft dysfunction after organ transplantation are closely related to the production of DSA, while NDSA and non HLA antibodies are responsible for graft survival. It has not been paid much attention to. It is reported that there is still another study on antibody mediated rejection mediated by antibodies against graft in the absence of DSA in the serum of organ transplant recipients. [4,5]., such as angiotensin receptor, anti wave protein antibody, anti oxidative lipid antibody, and so on in the body transplant patients. Antibodies are closely related to graft rejection and affect the long-term survival of the grafts in [7-10].. In addition to donor specific antibodies, non donor specific HLA antibodies and non HLA antibodies are also important factors affecting graft survival.
The antibody produced by organ transplant sensitization is mainly a specific anti donor HLA antibody. However, in the study, it is found that the serum of NDSA can also detect the emergence of [6,11-13] in the serum of the renal transplant recipients. The reasons for the production of NDSA are not completely clear on the premise of no specific immuno sensitization. The commonly accepted interpretation is the principle of cross reaction [14-17], That is, due to the existence of a common antigenic determinant among some of the HLA, the specific donor HLA antibody in the serum of the patients after renal transplantation can also be reacted with the HLA similar antigen expressed by the donor, thus showing the appearance of NDSA. However, some studies have found that the renal transplant patients can also be detected without DSA. Out of NDSA, the average median time of NDSA after renal transplantation was even earlier than that of DSA[14].. This phenomenon obviously can not be explained by the existing cross reaction theory.
Natural antibody (Natural antibodies) is a kind of unimmunized and widely existed antibody [18]. natural antibody response to various antigens that are not related to antigenic determinants. Therefore, it is characterized by multi reactivity and plays an important role in human innate immunity. The feature is to be able to combine [19-22]. with apoptotic cells
For a long time, the study of multi reactive antibodies is mainly concentrated in the field of human autoimmune diseases. However, the study of multi reactive antibody in the field of organ transplantation is rare. In view of the characteristics of the multi reactive antibody, we have explored the level of monoclonal B, the level of antibody and the serological level of renal transplant patients. The correlation between multiple reactive antibodies and the production of HLA antibodies. The correlation between the prestored multiple reactive antibodies in the renal transplant recipients and the antibody mediated rejection of the renal allograft and the long-term survival of the renal allograft were investigated.
1. Correlation between polyreactive antibody and HLA antibody production.
In this study, we selected and immortalized B lymphocytes from peripheral blood and surgical excised kidneys of renal transplant recipients and successfully produced monoclonal B lymphocyte clones. Through detection, we found that all B lymphocyte clones secreted antibodies, and some monoclonal anti bodies could be associated with a variety of antigens such as LPS, dsDNA, insulin, The antigen antibody reaction of the cell lysis product, which reveals the multi reactivity characteristic of the monoclonal antibody, is different from the specific antigen antibody reaction of "a key opening a lock". In addition, we have found that the multi reactive antibody can combine with the apoptotic cells. The specific monoclonal antibodies can react with a variety of HLA antigens, thus finding the correlation between the multi reactive antibody and the HLA antibody at the monoclonal antibody level. When the study was extended to the serological level of the patient, we found that the activity of the serum multi reactive antibody in the patients with HLA antibody positive was significantly higher than that of the patients with the negative HLA antibody. Increase (p=0.009, p0.001, p0.001). Application of Spearman correlation analysis found that there was a significant correlation between the activity of multi reactive antibody and the activity of HLA antibody in 300 patients with renal transplantation (p=0.004, p0.001, p=0.019). In patients with positive HLAI antibody, serum multi reactivity resistance was compared with those with low serum reactivity antibody. Patients with high body activity can react with more HLAI class antigens (p=0.030), thus revealing the correlation between reactive antibodies and HLA antibodies from serological level.
2, the correlation between preoperative serum reactivity antibody and renal transplant antibody rejection and long term survival of renal transplant recipients.
In 300 cases of renal transplant patients, we found that the activity of serum multi reactive antibody IgG in patients with renal transplantation was significantly higher than that in the normal population (p=0.011), and this difference was not due to the difference in the serum level of IgG in the renal transplant patients and the normal population. In this retrospective study, 300 patients followed. The time of visits was 81.2 + 35.3 months after the observation period of.46 patients with renal allograft dysfunction. We found that the serum reactivity IgG activity in patients with renal allograft dysfunction was significantly higher than that of normal renal transplantation (p0.001). And in 46 patients with renal allograft dysfunction, antibody mediated rejection resulted in renal allograft dysfunction. The patients' serum reactivity IgG activity was significantly higher than that of patients with other causes of graft dysfunction (p=0.033).Kaplan-Meier survival curve. The survival rate of renal allograft in patients with high reactive antibody activity before transplantation was significantly lower (p0.001), while multi reactive IgG was independent of HLA antibody to the survival of transplanted kidney. Further analysis of Cox proportional hazards model showed that IgG activity of serum reactive reactivity was an independent risk factor for renal allograft dysfunction (Hazard ratio=2.271, p0.001).
This study found that the reaction of serum multi reactive IgG antibody to apoptotic cells in renal transplantation patients is mainly mediated by IgG1 subtype and IgG3 subtype. The reaction of multi reactive antibody and apoptotic cells can activate complement pathway and cause C4d to be deposited on the surface of target cells. Further analysis shows that the responsiveness of IgG1 and IgG3 subtypes to the apoptotic cells and C4d are in the target. There was a significant correlation between cell surface deposition intensity (p0.001, p=0.005).
In this study, we found a significant correlation between the production of multiple reactive antibodies and the production of HLA antibodies from the level of monoclonal antibodies and the serological level of renal transplant recipients. In addition to the theory of cross reaction, a new basis for the cause of HLA antibody, especially NDSA, in organ transplant patients was provided, and the multiple reactive antibody IgG and renal transplantation were found. This study provides a new basis for the preoperative risk assessment of renal transplant recipients. In addition to the routine detection of HLA antibodies before organ transplantation, the monitoring of serum multi reactive antibodies is beneficial to the improvement of graft dysfunction after organ transplantation. The accuracy of the risk assessment provides a new theoretical and experimental basis for anti antibody therapy in high-risk patients before organ transplantation.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R699.2
[Abstract]:Antibody mediated graft rejection is the main cause of graft dysfunction and is also a key factor affecting the long-term survival of the transplanted kidney. It is widely believed that antibody mediated rejection and graft dysfunction after organ transplantation are closely related to the production of DSA, while NDSA and non HLA antibodies are responsible for graft survival. It has not been paid much attention to. It is reported that there is still another study on antibody mediated rejection mediated by antibodies against graft in the absence of DSA in the serum of organ transplant recipients. [4,5]., such as angiotensin receptor, anti wave protein antibody, anti oxidative lipid antibody, and so on in the body transplant patients. Antibodies are closely related to graft rejection and affect the long-term survival of the grafts in [7-10].. In addition to donor specific antibodies, non donor specific HLA antibodies and non HLA antibodies are also important factors affecting graft survival.
The antibody produced by organ transplant sensitization is mainly a specific anti donor HLA antibody. However, in the study, it is found that the serum of NDSA can also detect the emergence of [6,11-13] in the serum of the renal transplant recipients. The reasons for the production of NDSA are not completely clear on the premise of no specific immuno sensitization. The commonly accepted interpretation is the principle of cross reaction [14-17], That is, due to the existence of a common antigenic determinant among some of the HLA, the specific donor HLA antibody in the serum of the patients after renal transplantation can also be reacted with the HLA similar antigen expressed by the donor, thus showing the appearance of NDSA. However, some studies have found that the renal transplant patients can also be detected without DSA. Out of NDSA, the average median time of NDSA after renal transplantation was even earlier than that of DSA[14].. This phenomenon obviously can not be explained by the existing cross reaction theory.
Natural antibody (Natural antibodies) is a kind of unimmunized and widely existed antibody [18]. natural antibody response to various antigens that are not related to antigenic determinants. Therefore, it is characterized by multi reactivity and plays an important role in human innate immunity. The feature is to be able to combine [19-22]. with apoptotic cells
For a long time, the study of multi reactive antibodies is mainly concentrated in the field of human autoimmune diseases. However, the study of multi reactive antibody in the field of organ transplantation is rare. In view of the characteristics of the multi reactive antibody, we have explored the level of monoclonal B, the level of antibody and the serological level of renal transplant patients. The correlation between multiple reactive antibodies and the production of HLA antibodies. The correlation between the prestored multiple reactive antibodies in the renal transplant recipients and the antibody mediated rejection of the renal allograft and the long-term survival of the renal allograft were investigated.
1. Correlation between polyreactive antibody and HLA antibody production.
In this study, we selected and immortalized B lymphocytes from peripheral blood and surgical excised kidneys of renal transplant recipients and successfully produced monoclonal B lymphocyte clones. Through detection, we found that all B lymphocyte clones secreted antibodies, and some monoclonal anti bodies could be associated with a variety of antigens such as LPS, dsDNA, insulin, The antigen antibody reaction of the cell lysis product, which reveals the multi reactivity characteristic of the monoclonal antibody, is different from the specific antigen antibody reaction of "a key opening a lock". In addition, we have found that the multi reactive antibody can combine with the apoptotic cells. The specific monoclonal antibodies can react with a variety of HLA antigens, thus finding the correlation between the multi reactive antibody and the HLA antibody at the monoclonal antibody level. When the study was extended to the serological level of the patient, we found that the activity of the serum multi reactive antibody in the patients with HLA antibody positive was significantly higher than that of the patients with the negative HLA antibody. Increase (p=0.009, p0.001, p0.001). Application of Spearman correlation analysis found that there was a significant correlation between the activity of multi reactive antibody and the activity of HLA antibody in 300 patients with renal transplantation (p=0.004, p0.001, p=0.019). In patients with positive HLAI antibody, serum multi reactivity resistance was compared with those with low serum reactivity antibody. Patients with high body activity can react with more HLAI class antigens (p=0.030), thus revealing the correlation between reactive antibodies and HLA antibodies from serological level.
2, the correlation between preoperative serum reactivity antibody and renal transplant antibody rejection and long term survival of renal transplant recipients.
In 300 cases of renal transplant patients, we found that the activity of serum multi reactive antibody IgG in patients with renal transplantation was significantly higher than that in the normal population (p=0.011), and this difference was not due to the difference in the serum level of IgG in the renal transplant patients and the normal population. In this retrospective study, 300 patients followed. The time of visits was 81.2 + 35.3 months after the observation period of.46 patients with renal allograft dysfunction. We found that the serum reactivity IgG activity in patients with renal allograft dysfunction was significantly higher than that of normal renal transplantation (p0.001). And in 46 patients with renal allograft dysfunction, antibody mediated rejection resulted in renal allograft dysfunction. The patients' serum reactivity IgG activity was significantly higher than that of patients with other causes of graft dysfunction (p=0.033).Kaplan-Meier survival curve. The survival rate of renal allograft in patients with high reactive antibody activity before transplantation was significantly lower (p0.001), while multi reactive IgG was independent of HLA antibody to the survival of transplanted kidney. Further analysis of Cox proportional hazards model showed that IgG activity of serum reactive reactivity was an independent risk factor for renal allograft dysfunction (Hazard ratio=2.271, p0.001).
This study found that the reaction of serum multi reactive IgG antibody to apoptotic cells in renal transplantation patients is mainly mediated by IgG1 subtype and IgG3 subtype. The reaction of multi reactive antibody and apoptotic cells can activate complement pathway and cause C4d to be deposited on the surface of target cells. Further analysis shows that the responsiveness of IgG1 and IgG3 subtypes to the apoptotic cells and C4d are in the target. There was a significant correlation between cell surface deposition intensity (p0.001, p=0.005).
In this study, we found a significant correlation between the production of multiple reactive antibodies and the production of HLA antibodies from the level of monoclonal antibodies and the serological level of renal transplant recipients. In addition to the theory of cross reaction, a new basis for the cause of HLA antibody, especially NDSA, in organ transplant patients was provided, and the multiple reactive antibody IgG and renal transplantation were found. This study provides a new basis for the preoperative risk assessment of renal transplant recipients. In addition to the routine detection of HLA antibodies before organ transplantation, the monitoring of serum multi reactive antibodies is beneficial to the improvement of graft dysfunction after organ transplantation. The accuracy of the risk assessment provides a new theoretical and experimental basis for anti antibody therapy in high-risk patients before organ transplantation.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R699.2
【共引文獻(xiàn)】
相關(guān)期刊論文 前10條
1 陳新輝;趙明玄;安金剛;李巍;劉玉峰;;單克隆天然抗角蛋白抗體IgM 3B4體外抑制白念珠菌芽管形成及粘附作用[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2008年01期
2 隋燕霞;孫濤;趙東利;侯軍;李曉峰;楊U
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