PTA1（CD226）与肾移植患者急性排斥反应相关性的研究
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摘要
移植物排斥反应的判别在临床工作中有着非常重要的意义，通常判别排斥反应的最佳方式为病理活检，但这种方式是一种有创检查，不易反复进行，有出现严重并发症的危险性，且费用较高，患者不易接受。寻找一种渐变的，无创的、重复性好、费用较低的检测方式是临床工作的迫切需要。血小板/T细胞活化抗原1(platelet and T cell activation antigen 1，PTA1，CD226)是1997年基因克隆成功的新分子，同许多免疫细胞膜型分子一样存在着可溶性形式及膜型分子。对外周血中活化细胞分子这两种分子形式的检测可以达到判别移植物排斥反应的目地。
观察尸体肾移植术后患者急性排斥反应时血清sPTA1水平及细胞膜mPTA1的表达与排斥反应的相关性，使之成为移植物排斥反应的判别指标。选取了17例围手术期同种尸体肾移植患者以及2例术后超过1年的急性排斥
＄－fkkq4＋qtwt
一
反应的患者，根据患者的不同病情状况采取血样，采用夹心ELISA法狈定患
者血清slunl水平，采用流式细g淋巴细胞上wtAI表达，对明确
有排斥反应、可F斥反应以及不能区分排斥反应的患者在B超5！导下行
移植’肾活检穿刺，行光镜、电镜及免疫组化检查。
门例围手术期患者术前sPTAI水平及InPTAI ＄jA阳性率与正常对照接
近，无显著差异。术后第1天sPTA即有高水平的表达，与对照组差异显著。
19例尸体‘肾移植患者中经病理证实的5例排斥反应患者sPTAI显著升高，
InPTAI ＆jk增强，与对照组差异非常显著。经加强免u剂治疗后，血清
sPTAI下降，InPTAI ＆：IH下降。而且sPTAI水平的变化早于临床症状表现，
持续时间较长。5例排斥反应的患者中病理学检查证实：4例为细【斥
反应；l例为血管 诉a，日后发展为慢性排斥反应。
移植F斥反应时的T细胞活化、增殖是一个渐进的过程，只有T细胞
克隆扩增到一定数量，在移植物内大量积聚，并对靶细胞发挥明显的破坏作用，
才出现临床表现和病理学变化。PTA水平变化早于临床表现和病理学变化；
PTAI是一项较为可靠的、无仓的、重复性好、费用较低的判别和监R梆斥反
应的指标，与病理检查的结果有较好的一致性，可以作为移F斥反应的监
测和判别指标。
It is important to distinguish the allograft actue rejection in clinic . Generally , the best way to distinguish and monitor the episodes of allograft acute rejection is biopsy . But this way is invasive, bears the risk of potentially dangerous complication, costly and not being easily accepted by patients. Therefore, the availability of a sensitive, specifiable, non-invasive screening for the detection of allograft acute rejection would be highly desireabl to facilitate the task in clinic. Platelet and T cell activation antigen 1 ( PTA1 , CD226 ) is a novel immuno activated molecul that being cloned
successfully in 1997, like the other membran molecul PTA1 have two form of solubl and membranous .To study the interrelationship between PTA1 and allograft actue rejection in renal transplantation , to make it become the probe to distinguish allograft actue rejection. When five cases of 19 after renal transplant recipiebts were distinguished as allograft actue rejection by histopathologic examination , according to respective condition of recipients take the sample of blood , Solid-phase ligand ELISA method was used to analyzed serum of sPTAl in renal transplant recipients and Flow cytometry was emploied to study the expression of PTA1 on h/mphtic cells. The level of serum sPTAl had increased remarkably , the expression of mPTAl on lymphtic cells had enhanced ,and the change of the level of serum sPTAl occurred earlier than clinic symptom and that in histopathology , it had decreased rapidly after enhancing the treatment of immune .The activation of infiltration T cell in the allograft of renal transplantation was a gradual process . The allograft did not show signs of acute rejection to the clinic symptom and/or histopathology under the microscope until the activation reached to a certain level. PTA1 is one of sensitive , specific , non-invasive , repeatable , cheap and creditable guideline to distinguish and monitor for allograft renal transplant recipients , the result is consistent with histopathological examination.
观察尸体肾移植术后患者急性排斥反应时血清sPTA1水平及细胞膜mPTA1的表达与排斥反应的相关性，使之成为移植物排斥反应的判别指标。选取了17例围手术期同种尸体肾移植患者以及2例术后超过1年的急性排斥
＄－fkkq4＋qtwt
一
反应的患者，根据患者的不同病情状况采取血样，采用夹心ELISA法狈定患
者血清slunl水平，采用流式细g淋巴细胞上wtAI表达，对明确
有排斥反应、可F斥反应以及不能区分排斥反应的患者在B超5！导下行
移植’肾活检穿刺，行光镜、电镜及免疫组化检查。
门例围手术期患者术前sPTAI水平及InPTAI ＄jA阳性率与正常对照接
近，无显著差异。术后第1天sPTA即有高水平的表达，与对照组差异显著。
19例尸体‘肾移植患者中经病理证实的5例排斥反应患者sPTAI显著升高，
InPTAI ＆jk增强，与对照组差异非常显著。经加强免u剂治疗后，血清
sPTAI下降，InPTAI ＆：IH下降。而且sPTAI水平的变化早于临床症状表现，
持续时间较长。5例排斥反应的患者中病理学检查证实：4例为细【斥
反应；l例为血管 诉a，日后发展为慢性排斥反应。
移植F斥反应时的T细胞活化、增殖是一个渐进的过程，只有T细胞
克隆扩增到一定数量，在移植物内大量积聚，并对靶细胞发挥明显的破坏作用，
才出现临床表现和病理学变化。PTA水平变化早于临床表现和病理学变化；
PTAI是一项较为可靠的、无仓的、重复性好、费用较低的判别和监R梆斥反
应的指标，与病理检查的结果有较好的一致性，可以作为移F斥反应的监
测和判别指标。
It is important to distinguish the allograft actue rejection in clinic . Generally , the best way to distinguish and monitor the episodes of allograft acute rejection is biopsy . But this way is invasive, bears the risk of potentially dangerous complication, costly and not being easily accepted by patients. Therefore, the availability of a sensitive, specifiable, non-invasive screening for the detection of allograft acute rejection would be highly desireabl to facilitate the task in clinic. Platelet and T cell activation antigen 1 ( PTA1 , CD226 ) is a novel immuno activated molecul that being cloned
successfully in 1997, like the other membran molecul PTA1 have two form of solubl and membranous .To study the interrelationship between PTA1 and allograft actue rejection in renal transplantation , to make it become the probe to distinguish allograft actue rejection. When five cases of 19 after renal transplant recipiebts were distinguished as allograft actue rejection by histopathologic examination , according to respective condition of recipients take the sample of blood , Solid-phase ligand ELISA method was used to analyzed serum of sPTAl in renal transplant recipients and Flow cytometry was emploied to study the expression of PTA1 on h/mphtic cells. The level of serum sPTAl had increased remarkably , the expression of mPTAl on lymphtic cells had enhanced ,and the change of the level of serum sPTAl occurred earlier than clinic symptom and that in histopathology , it had decreased rapidly after enhancing the treatment of immune .The activation of infiltration T cell in the allograft of renal transplantation was a gradual process . The allograft did not show signs of acute rejection to the clinic symptom and/or histopathology under the microscope until the activation reached to a certain level. PTA1 is one of sensitive , specific , non-invasive , repeatable , cheap and creditable guideline to distinguish and monitor for allograft renal transplant recipients , the result is consistent with histopathological examination.
引文
[1] 金伯泉，主编.细胞和分子免疫学[M].北京：科学出版社，2001．
[2] 黎磊石.中国终末期肾病.肾脏病与透析肾移植杂志.1995；4：77
[3] Cohen DJ.Loertscher R,Rubin MF.et al. Cyclosporine: A new immunosuppressive agent for organ transplantation. Ann Int Med 1984:101:667-682
[4] ROCHER LLMLFORD EL. KIRKMAN RL .et al. Conversion from Cyclosporine to Azathioprine in renal allograflrecipients.Transplantation 1984:38;669-674
[5] BILLINGHAM ME Diagnosis of cardiac rejection by endomyocardial55 biopsy.Heart Transplant 1982:1:25-30
[6] HAVERICH A KEMNTTZ J, FIEGUTH HG, et al .Non-invasive parameters for detection of cardiac allograft rejections. Clin Transplant 1987;1:151-158
[7] Bums GF, Triglia T, Werkmeister-JA, et al. TliSAl. a human T lineage-specific activation antigen involved in the differentiation of cytotoxic T lymphocytes and anomalous killer cells from their precursors. J Exp Med,1985,161:1063-1078．
[8] Sherrington PD, Scott JL, Jin B, et al.TliSAl(PTAl) activation antigen implicated in T cell differentiation and platelet activation is a member of the immunoglobulin superfamily exhibiting distinctive regulation of expression. J Bio Chem, 1997,272:21735-21744．
[9] 孙凯，冯琦，金伯泉.血小板/T细胞活化抗原1． 细胞和分子免疫学杂
志，1999，19(3)：261-264
[10] Chen LK, Bensusan A, Burns GF, et al.Tourrvieille-B;SA1-lospecific prooliiiferative human T-cell clones aacquire the cytotoxic effector function after three months in culture, in IL-2 conditioned medium. Hum Immunol, 1986,17:30-36.
[11] Jin B, Scott JL. Vadas MA, et al. TGF beta down-regulates TliSA1 expression and inhibits the differentiation of precursor lymphocytes into CTL and LAK cells. Immunology, 1989,66:570-576.
[12] Scott JL, Dunn SM, Jin B, et al. Characteritation of a novel membrane glycoprotein involved in platelet activation. J Biol Chem, 1989, 264:13475-13482.
[13] Zuzel, Walton S, Bnrns GF, et al. A monoclonal antibody to a 67kD cell membrane gllycoprotein directly induces persistent platelet aggregation independently of granule secretion. Bfit J Haematol, 1991,79:466-473.
[14] 黄传书，金伯泉，汪美先，等．HFRS患者PBMC和异型淋巴细胞多种活化抗原表达的研究．上海免疫学杂志，1991，11：94-97．
[15] 王坚，胡绍文，王泊云，等．自身免疫性加装病浸润T细胞活化与滤泡细胞DR抗原表达．中华医学杂志，1992，27：77-80．
[16] Tabata H, Hara M, Kitani A, et al. Hirose-T expression of TliSA1 on Tcells from patients with rheumatoid arthritis and systemic lupus erythematosus. Clin Immunol Immunopathol, 1989,52:366-375.
[17] Miller FW, Love LA, Barbieri SA, et al. Lymphocyte activation markers in idiopathic myositis: changes with disease activity and differences among clinical and autoantibody subgroups. Clin Exp Immunol, 1990,81:373-379.
[18] Huang C, Jin B, wang M, et al. Hemorrhagic fever with renal
syndrome:relationship betweet pathogenesis and celluar immunity. J Infect Dis, 1994,169: 868-870．
[19] 贾卫，金伯泉，李琦，等.检测可溶性PTA1分子夹心ELISA方法的建立细胞和分子免疫学杂志2000, 16 (4) : 282-284．
[21] ERTELW REICHENSPURNER H iERSCH C, et al. Cytommunological monitoring in acute rejection and viral. Bacterial or fungal infection followingtransplantation .Heart Transplant 1985:4:309-393
[22] Loertscher R, Forbes BD. Halabi G, et al. Expression of early and activation markers on peripheral blood T lymphocytes does not reliably reflect immune events in transplanted hearts [J].CIin transplant, 1994,8(3) :230-238．
[23] COTNER T. WILLIAMS JM, CHRISTENSON L. et al. Simultaneous flow cytometric analysis of human T cell activation antigen expression and DNA content J EXPMed 1983;157:46M72．
[24] WILLIAMSJM, LOERTSCHER R, COTNER T. et al. Dual parameter flow cytometric analysis of DNA content ,activation antigen expression and T cell subset proliferation in the humanmixed lymphocyte reaction. J Immunol 1984:132:2330-2337．
[25] WINDSOR NT, LLOYD KS. YOUNG JB, et al. Dynamics of soluble Interleukin-2 receptor levels immediately after heart transplantation. Transplantation 1991:52:78-82
[26] HUEBERS HAWFINCH CA.The physiology of transferrin and transferrin receptors.Physiol Reviews 1987:67:520-582．
[27] BURNS GF. WERKMEISTER JA,TRIGLIA T.High frequencies of precursors of anomalous killer cells in human peripheral
blood:evidence for T cell regulation. Cell Immunol 1984:89:202-211.
[28] HEMLER ME, JACOBSON JG, BRENNER MB, et al.VLA-1:aT cell surface antigen which defines a novel Iate stage of human T cell activation. Eur J Immunol 1985:15:502-508.
[29] 董光龙，王为忠，吴国生，等：血清sPTA1与人活体小肠移植急性排斥反应的相关性评价[J]．细胞和分子免疫学杂志，2000，16(6)：439-441．
[30] 田方，金伯泉，姜绍谆，等．一种新的免疫球蛋白超家族成员PTA1的细胞分布及表达的研究。细胞和分子免疫学杂志，1996，12：53-55
[31] 李德敏，金伯泉，苏丽，等．小鼠血小板／T细胞活化抗原1的表达分布与功能．细胞和分子免疫学杂志，1999，19(3)：232—235．
[32] 陈丽华，金伯泉，贾卫，等．一种新的人内皮细胞可诱导性粘附分子PTA1的表达、调变及粘附功能的研究．细胞和分子免疫学杂志，2000，16(1)：21—25．
[33] Bussolino F, ZicheM, Wang JM et al. In vitroand in vivo activation of endothelial cellsby colony-stimulating factors [J]. J Clin Invest, 1991;87:986-995.
[34] CarrMW, Alon R, Springer TA. The c-c chemokine MCP-1 differentially modulates the avidity of beta 1 and beta2 integrins on T Iymphocytes[J]. Immunity, 1996;4(2):179-87.
[35] Read MA,NeishAS,LuscinskasFW et al. The proteasome pathway is required for cytokine-induced endothelial-Ieukocyte adhesion molecule expression[J]. Immunity, 1995;2(5):493-506.
[36] Zimmeramn GA, Mcintyre TM, Prescott SM. Thrombin stimulatesthe adherence of neutrophils to human endothelial cells in vitro[J]. J Clin Invest, 1985;76:2235.
[37] 佘恒，陈丽华，金伯泉．PTA1参与活化内皮细胞和淋巴细胞间粘附的形态学观察．细胞和分子免疫学杂志，2000，16(2)：138．
[38] Shibuya K, Lanier LL, Phillips JH, et al. Physical and function association of LFA-1 with DNAM-1 adhesion molecule. Immunity,1999:11:615-623
[39] 李州利，孙凯，张继帅，等．两种新发现的与移植免疫有关的白细胞分化抗原．肾脏病与透析肾移植杂志．2002．
[40] 杨俊伟．移植肾病理学及与临床的关系．肾脏病与透析肾移植杂志．1996；5：86
[41] KAYE MAP. The registry of the International Society For Heart and Lung Transplantation Ninth Official report 1992.J Heart Lung Transplant 1992;11;599—606
[42] Suthanthiran M, strom TB. Renal transplantation. N Engl J Med, 1994;331:365
[2] 黎磊石.中国终末期肾病.肾脏病与透析肾移植杂志.1995；4：77
[3] Cohen DJ.Loertscher R,Rubin MF.et al. Cyclosporine: A new immunosuppressive agent for organ transplantation. Ann Int Med 1984:101:667-682
[4] ROCHER LLMLFORD EL. KIRKMAN RL .et al. Conversion from Cyclosporine to Azathioprine in renal allograflrecipients.Transplantation 1984:38;669-674
[5] BILLINGHAM ME Diagnosis of cardiac rejection by endomyocardial55 biopsy.Heart Transplant 1982:1:25-30
[6] HAVERICH A KEMNTTZ J, FIEGUTH HG, et al .Non-invasive parameters for detection of cardiac allograft rejections. Clin Transplant 1987;1:151-158
[7] Bums GF, Triglia T, Werkmeister-JA, et al. TliSAl. a human T lineage-specific activation antigen involved in the differentiation of cytotoxic T lymphocytes and anomalous killer cells from their precursors. J Exp Med,1985,161:1063-1078．
[8] Sherrington PD, Scott JL, Jin B, et al.TliSAl(PTAl) activation antigen implicated in T cell differentiation and platelet activation is a member of the immunoglobulin superfamily exhibiting distinctive regulation of expression. J Bio Chem, 1997,272:21735-21744．
[9] 孙凯，冯琦，金伯泉.血小板/T细胞活化抗原1． 细胞和分子免疫学杂
志，1999，19(3)：261-264
[10] Chen LK, Bensusan A, Burns GF, et al.Tourrvieille-B;SA1-lospecific prooliiiferative human T-cell clones aacquire the cytotoxic effector function after three months in culture, in IL-2 conditioned medium. Hum Immunol, 1986,17:30-36.
[11] Jin B, Scott JL. Vadas MA, et al. TGF beta down-regulates TliSA1 expression and inhibits the differentiation of precursor lymphocytes into CTL and LAK cells. Immunology, 1989,66:570-576.
[12] Scott JL, Dunn SM, Jin B, et al. Characteritation of a novel membrane glycoprotein involved in platelet activation. J Biol Chem, 1989, 264:13475-13482.
[13] Zuzel, Walton S, Bnrns GF, et al. A monoclonal antibody to a 67kD cell membrane gllycoprotein directly induces persistent platelet aggregation independently of granule secretion. Bfit J Haematol, 1991,79:466-473.
[14] 黄传书，金伯泉，汪美先，等．HFRS患者PBMC和异型淋巴细胞多种活化抗原表达的研究．上海免疫学杂志，1991，11：94-97．
[15] 王坚，胡绍文，王泊云，等．自身免疫性加装病浸润T细胞活化与滤泡细胞DR抗原表达．中华医学杂志，1992，27：77-80．
[16] Tabata H, Hara M, Kitani A, et al. Hirose-T expression of TliSA1 on Tcells from patients with rheumatoid arthritis and systemic lupus erythematosus. Clin Immunol Immunopathol, 1989,52:366-375.
[17] Miller FW, Love LA, Barbieri SA, et al. Lymphocyte activation markers in idiopathic myositis: changes with disease activity and differences among clinical and autoantibody subgroups. Clin Exp Immunol, 1990,81:373-379.
[18] Huang C, Jin B, wang M, et al. Hemorrhagic fever with renal
syndrome:relationship betweet pathogenesis and celluar immunity. J Infect Dis, 1994,169: 868-870．
[19] 贾卫，金伯泉，李琦，等.检测可溶性PTA1分子夹心ELISA方法的建立细胞和分子免疫学杂志2000, 16 (4) : 282-284．
[21] ERTELW REICHENSPURNER H iERSCH C, et al. Cytommunological monitoring in acute rejection and viral. Bacterial or fungal infection followingtransplantation .Heart Transplant 1985:4:309-393
[22] Loertscher R, Forbes BD. Halabi G, et al. Expression of early and activation markers on peripheral blood T lymphocytes does not reliably reflect immune events in transplanted hearts [J].CIin transplant, 1994,8(3) :230-238．
[23] COTNER T. WILLIAMS JM, CHRISTENSON L. et al. Simultaneous flow cytometric analysis of human T cell activation antigen expression and DNA content J EXPMed 1983;157:46M72．
[24] WILLIAMSJM, LOERTSCHER R, COTNER T. et al. Dual parameter flow cytometric analysis of DNA content ,activation antigen expression and T cell subset proliferation in the humanmixed lymphocyte reaction. J Immunol 1984:132:2330-2337．
[25] WINDSOR NT, LLOYD KS. YOUNG JB, et al. Dynamics of soluble Interleukin-2 receptor levels immediately after heart transplantation. Transplantation 1991:52:78-82
[26] HUEBERS HAWFINCH CA.The physiology of transferrin and transferrin receptors.Physiol Reviews 1987:67:520-582．
[27] BURNS GF. WERKMEISTER JA,TRIGLIA T.High frequencies of precursors of anomalous killer cells in human peripheral
blood:evidence for T cell regulation. Cell Immunol 1984:89:202-211.
[28] HEMLER ME, JACOBSON JG, BRENNER MB, et al.VLA-1:aT cell surface antigen which defines a novel Iate stage of human T cell activation. Eur J Immunol 1985:15:502-508.
[29] 董光龙，王为忠，吴国生，等：血清sPTA1与人活体小肠移植急性排斥反应的相关性评价[J]．细胞和分子免疫学杂志，2000，16(6)：439-441．
[30] 田方，金伯泉，姜绍谆，等．一种新的免疫球蛋白超家族成员PTA1的细胞分布及表达的研究。细胞和分子免疫学杂志，1996，12：53-55
[31] 李德敏，金伯泉，苏丽，等．小鼠血小板／T细胞活化抗原1的表达分布与功能．细胞和分子免疫学杂志，1999，19(3)：232—235．
[32] 陈丽华，金伯泉，贾卫，等．一种新的人内皮细胞可诱导性粘附分子PTA1的表达、调变及粘附功能的研究．细胞和分子免疫学杂志，2000，16(1)：21—25．
[33] Bussolino F, ZicheM, Wang JM et al. In vitroand in vivo activation of endothelial cellsby colony-stimulating factors [J]. J Clin Invest, 1991;87:986-995.
[34] CarrMW, Alon R, Springer TA. The c-c chemokine MCP-1 differentially modulates the avidity of beta 1 and beta2 integrins on T Iymphocytes[J]. Immunity, 1996;4(2):179-87.
[35] Read MA,NeishAS,LuscinskasFW et al. The proteasome pathway is required for cytokine-induced endothelial-Ieukocyte adhesion molecule expression[J]. Immunity, 1995;2(5):493-506.
[36] Zimmeramn GA, Mcintyre TM, Prescott SM. Thrombin stimulatesthe adherence of neutrophils to human endothelial cells in vitro[J]. J Clin Invest, 1985;76:2235.
[37] 佘恒，陈丽华，金伯泉．PTA1参与活化内皮细胞和淋巴细胞间粘附的形态学观察．细胞和分子免疫学杂志，2000，16(2)：138．
[38] Shibuya K, Lanier LL, Phillips JH, et al. Physical and function association of LFA-1 with DNAM-1 adhesion molecule. Immunity,1999:11:615-623
[39] 李州利，孙凯，张继帅，等．两种新发现的与移植免疫有关的白细胞分化抗原．肾脏病与透析肾移植杂志．2002．
[40] 杨俊伟．移植肾病理学及与临床的关系．肾脏病与透析肾移植杂志．1996；5：86
[41] KAYE MAP. The registry of the International Society For Heart and Lung Transplantation Ninth Official report 1992.J Heart Lung Transplant 1992;11;599—606
[42] Suthanthiran M, strom TB. Renal transplantation. N Engl J Med, 1994;331:365