慈濟 陳政宏 開發 C4d及CR1比值 於診斷運用(紅斑狼瘡&心臟移植衰竭)!!

狼瘡生物指標　精準判定紅斑狼瘡鉅亨網新聞中心(來源：健康醫療網)2015-06-1614:01 （健康醫療網／記者郭庚儒報導）全身性紅斑狼瘡患者出現發燒症狀，究竟是疾病復發還是感染肺炎？臨床上，醫師遇到類似情形，總是很困擾；台北慈濟醫院研發「狼瘡生物指標」，只要抽血就能判定病程活躍度，及時給予患者準確治療，避免過度使用免疫抑制藥物，而產生敗血症的風險。台北慈濟醫院風濕免疫科主任陳政宏帶領研究團隊，研發出新單株抗體「CR1-2B11」，具有正確估算紅血球上第一型補體受體CR1的能力，進而用於紅斑性狼瘡患者紅血球上C4裂解物C4d及CR1的比值，作為「狼瘡生物指標」。針對87名紅斑性狼瘡患者，使用流式細胞儀分析後發現，當病程活躍時比值明顯上升，治療後病程得到控制時比值明顯下降。陳政宏指出，全身性紅斑狼瘡患者，常因病程活躍度或合併感染疑慮，造成醫師無法準確判斷而延誤病情，有了此項生物指標，患者只要經由抽血，2至3小時候即可判定狼瘡病程是否活躍，較現行臨床上使用檢驗項目，更具敏感性及專一性。陳政宏比喻，全身性紅斑狼瘡的病程演變多形多樣，就像齊天大聖孫悟空可72變，而「狼瘡生物指標」可比擬為如來佛掌，協助醫師鑑別診斷病程活躍度、評估用藥療效，並與感染作診斷區分，讓患者早日脫離紅斑性狼瘡對生命的威脅。

中文題目：使用單株抗體CR1-2B11測量紅血球上C4d與CR1比值於紅斑性狼瘡患者：臨床運用及結果英文題目：Measurement of Erythrocyte C4d to CR1 Ratio by Using CR1-2B11 in Systemic LupusErythematosus Patients: Clinical Applications and Results作 者：陳政宏 田炯璽 賴振宏 郭三元 侯宗昀 劉峰誠 林剛民 張德明服務單位：國防醫學院三軍總醫院風濕免疫過敏科 前言：測量紅血球上C4d及CR1 的比值，是一相當有潛能的生物指標(Biomarker)來預估狼瘡的病情活躍程度，並且能提供臨床醫師作出適當的治療決策，並且也有提早診斷紅斑性狼瘡的潛在能力；我們研發出一新的單株抗體(CR1-2B11)具有正確估算紅血球上CR1(Complement receptor 1) 的能力，因此運用此抗體來研究紅斑性狼瘡病患紅血球上C4d及CR1 的比值，並以此數值來比對狼瘡患者的病程，以了解此一生物指標於臨床運用之潛能。材料及方法：我們從六十八位已確定診斷為紅斑性狼瘡的住院患者，我們使用流式細胞儀分別運用CR1-2B1 及 anti-C4d抗體來研究病患紅血球上C4d及CR1 的消長並計算出此一生物指標的實際數值，並且分析與臨床病程之關聯性。結果： 從三十六位活躍紅斑性狼瘡病患所獲的比值為15.72 ± 7.78,非活躍紅斑性狼瘡但合併感染之病患之比值為3.26 ± 2.64,可見此生物指標明顯於狼瘡活躍時高於非活躍時之感染發燒，這有益於臨床醫師鑑別診斷因發燒而入院之紅斑性狼瘡病患；我們也追蹤了同數位狼瘡患者生物指標的變化，當病程活躍時其比值明顯上升，當治療後病程明顯得到控制時比值明顯下降；另外從三十二位活躍紅斑性狼瘡患者我們也追蹤其治療前與治療後之指標變化( 10.34 ±6.34 versus 3.27 ± 2.32; p < 0.0001). 結論：以上結果說明了運用此一新發展出的單株抗體(CR1-2B11)來計算紅血球上C4d及CR1所獲得的生物指標具有評估療效之能力，並且具有鑑別診斷發燒之紅斑性狼瘡病患是否合併有感染或病情活躍的潛能；同時也具有預估狼瘡的病情活躍程度的能力。

Antibody CR1-2B11 recognizes a non-polymorphic epitope of human CR1 (CD35). Clin Exp Immunol. 2007 Jun;148(3):546-54. Chen CH1, Ghiran I, Beurskens FJ, Weaver G, Vincent JA, Nicholson-Weller A, Klickstein LB.  1Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Boston, MA, USA.  Abstract  Measurement of erythrocyte [red blood cells (RBC)] complement receptor type 1 (CR1, CD35) has the potential to serve as a sensitive assessment of complement activation and immune complex clearance. All previously reported monoclonal antibodies (MoAb) to the extracellular region of CR1 recognize epitopes within the long homologous repeats (LHR) of CR1 and the epitopes for the most frequently used MoAbs are repeated at least twice per CR1 molecule. Furthermore, CR1 exhibits structural polymorphism characterized by a variable number of LHR per molecule. Thus, accurate enumeration of cell surface CR1 using currently available MoAb would require that the results be corrected for the number of antibody epitopes per CR1 molecule encoded by each individual's alleles. To obtain a MoAb to a non-polymorphic epitope on human CR1, hybridomas were generated from mice immunized with recombinant soluble CR1 (sCR1) and MoAb were screened for those that recognized the full-length extracellular domain but failed to bind to all four recombinant LHR fragments. A single antibody, CR1-2B11, was identified and was found to recognize an epitope located wholly within SCR29-30 of CR1, NH2-terminal to an elastase cleavage site. Like other CR1 MoAb, the CR1-2B11 epitope expression decreased on old erythrocytes compared to younger cells and CR1-2B11 did not identify a CR1 'stump' on RBC. Importantly, CR1-2B11 immunofluorescence did not change with storage or handling of RBC, unlike the apparent decrease in immunofluorescence observed with other MoAb. CR1-2B11 should be useful for the accurate enumeration of RBC CR1.

Measurement of human erythrocyte C4d to erythrocyte complement receptor 1 ratio in cardiac transplant recipients with acute symptomatic allograft failure. Transplant Proc. 2008 Oct;40(8):2638-42. BACKGROUND: Complement activation has been recognized as a contributing factor to cardiac allograft dysfunction. Combined measurement of erythrocyte C4d (E-C4d) and complement receptor 1 (E-CR1) are potential biomarkers to monitor complement activity in patients with autoimmune diseases. We conducted a prospective study using CR1-2B11 monoclonal antibody to detect the E-C4d to E-CR1 ratio among our cardiac transplant recipients with acute symptomatic allograft failure. MATERIALS AND METHODS: Eight recipients with acute cardiac allograft failure and 72 healthy controls were included in this study. Levels of E-C4d and E-CR1 were measured by indirect immunofluorescence and flow cytometry. The results were utilized to determine the association between patient C4d staining, histological features, and clinical outcomes. RESULTS: Eight patients with nine episodes of sudden onset of graft failure and suspected antibody-mediated rejection (AMR) were included in this study. One patient who received emergent mechanical circulatory support was treated with plasmapheresis for his unstable hemodynamic status. The mean pretreatment left ventricular ejection fraction was 30.3%. No histological study demonstrated cellular rejection or AMR in any patient. There were two patients with positive C4d immunostaining. Three patients had four episodes of acute rejection with sudden death at home. The mean E-C4d/E-CR1 ratio in the study group (n = 9) was 0.22 +/- 0.07, and 0.12 +/- 0.10 in the control group (n = 72). As comparing both groups, we found the ratios were significant higher in the study group (P = .0003). CONCLUSIONS: Measurement of the E-C4d/E-CR1 ratio may be a noninvasive method for detecting acute rejection after cardiac transplantation.

 造福狼瘡病患！台北慈院研發生物指標鑑別診斷 2015-06-1609:24 〔記者翁聿煌／新北報導〕27歲李小姐患有紅斑性狼瘡，因感冒發燒後，出現咳血、高燒不退及大量肺出血的症狀，嚴重到需使用葉克膜維持生命，在尚未確知是其他感染或病程復發時，台北慈濟醫院風濕免疫科主任陳政宏運用新研發的狼瘡生物指標予以輔助鑑別診斷，確定是病程復發後，便運用血漿置換術及類固醇脈衝療法及時搶救，7天後病患移除葉克膜，27天後順利出院。 全身性紅斑性狼瘡是一種自體免疫細胞自我攻擊的疾病，台灣罹患紅斑性狼瘡人數至少在1萬人以上，但全身性紅斑狼瘡因病程演變多形多樣，讓臨床醫師處理過程中很棘手，台北慈濟醫院風濕免疫科陳政宏主任與其研究團隊，研發出的「狼瘡生物指標」，在臨床上可協助醫師鑑別診斷病程活躍度、評估用藥療效；或與感染作診斷區分，讓患者早日脫離紅斑性狼瘡對生命的威脅。 陳政宏指出，紅斑性狼瘡每年發生的新增病例約為400至1600人，其中又以15至45歲的女性為好發族群，比例約佔90％。其臨床症狀小則關節痛、口腔黏膜潰瘍，大則引發內臟系統的攻擊，如心包膜炎、肺炎等嚴重症狀，甚至可能導致肺出血、多重器官血栓或侵犯中樞神經等致命的併發症，當這些症狀出現時，常會因合併感染疑慮或對狼瘡病程活躍度無法準確判斷而延誤病情。 陳政宏說，除了藥物治療外，紅斑性狼瘡是一種慢性疾病，目前雖然無法治癒，但只要與醫師合作，平時避免烈日照射，保持充足睡眠、適當運動、預防感染，患者5年存活率高達9成，並可過著宛如一般人的正常生活。