日本組織適合性学会誌第23巻1号

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1 Major Histocompatibility Complex 2016; 23 (1): 総説 1) 1) 1) 1) 2) 2) 2) 2) 2) 2) 1) 2) ABO A/B A/B-IgG A/B-IgG A/B-IgG ABO キーワード : A/B A/B FCM-C1q FCM-IgG はじめに ABO CD20 Rituximab: Genetech, San Francisco, CA, USA 1) A/B DFPP: Double Filtration Plasmapheresis PE: Plasma Exchange HLA HLA-IgG DSA: donor specific alloantibody DSA 2) A/B-IgG C1q FCM-C1q: flow cytometry method for complement C1q test I. 対象および方法 ABO A B O 10 A B O 10 A/B FCM-IgG: flow cytometry method for IgG test FCM-C1q ABO CDC-XM: Complement-Dependent Cytotoxic Crossmatch test FCXM-IgG: flow cytometry lym TEL: FAX: ishizuka.tsutomu@twmu.ac.jp 39

2 MHC 2016; 23 (1) phocyte crossmatch-igg test LABScreen: LABScreen single antigen One Lambda Inc., Canoga Park, CA, USA DSA 6 A/B ID-AGT: Indirect anti-globulin test FCM-C1q JMP Pro Wilcoxon rank sum test 5% 1)flow cytometry method for complement C1q test A/B-IgG A/B-IgM 5 mm DTT: dithiothreitol 37 C 30 3,4) AB Sigma-Aldrich, St. Louis, Mo, United States DTT A/B-IgG 50 µl DiaCell ABO A1-B DiaMed AG, Switzerland 50 µl 30 Complement component C1q from human serum Sigma-Aldrich 5 µl 20 Dako REAL Antibody Diluent Dako Denmark A/S Code: S2022 Anti-Human-C1q-FITC Medical and Biological Laboratories Co. Ltd µl 20 FACS- Calibur HG Becton Dicknson and Company, SanJose, CA, USA FCM-C1q % 2)flow cytometry method for IgG test A/B-IgG A/B- IgM 5 mm DTT 37 C 30 DTT AB DTT A/B-IgG 50 µl ABO A1-B 50 µl 30 fluorescein-isothiocyanate (FITC)-conjugated anti-human IgG Jackson Immunoresearch Laboratories, West Grove, Penn, United States µl 20 FACSCalibur HG FCM-IgG 2 n 5) II. 結果 1) 健常者および透析患者の血液型抗 A/B 抗体の検討 O 7:3 52.6± A 5:5 55.0± B 6:4 55.4± O 8:2 55.8± A 9:1 58.5± B 7:3 49.1± A/B FCM- IgG FCM-C1q FCM-IgG 2 n O A 6.4±2.1 図 1 A/B A B O 5% FCM-IgG FCM-C1q 40

3 MHC 2016; 23 (1) 表 1 ABO No. ID-AGT 2 n FCM-C1q % 1 A O A O B O A O A O A B 図 2 ABO A/B ID-AGT FCM-C1q 3 11 B 5.1± A B 1.9± B A 2.4± FCM-C1q % O A 27.2± B 16.4± A B 1.7± B A 0.8± FCM-IgG 2 n O A 6.0± B 5.5± A B 1.8± B A 2.4± FCM-C1q %O A 28.3± B 22.1± A B 8.6± B A 3.5± )ABO 血液型不適合生体腎移植を施行した 6 症例 1 ABO ID-AGT 2 n 1 6 FCM-C1q ID-AGT III. 考察 Alexandre GP ABO 6,7) 1989 ABO Rituximab ABO ABO 8) 41

4 MHC 2016; 23 (1) 9) A/B ID-AGT MTS: DiaMed-ID Micro Typing System, DiaMed AG, Switzerland CAT: BioVue Columm Agglutination Technology, Ortho-Clinical-Diagnostics, Tokyo, JPN flow cytometry IgG IgG1 4 A/B-IgG A/B-IgG 10) A B O 5% A/B-IgG FCM-C1q O A B A/B- IgG FCM-C1q O A B 5% preliminary study A/B-IgG 11,12) A/B-IgG A/B-IgG IgG IgG1 60% IgG2 29% IgG3 7% IgG4 4% IgG1, IgG3 IgG2 IgG4 13) FCM-C1q A B A/B IgG4 IgG IgG4 FCM-C1q IgG data not shown A/B 14) A B A/B O A/B FCM-C1q A B A/B-IgG ABO O A B O 15) A/B-IgG ABO 6 ID- AGT FCM-C1q ID-AGT FCM-C1q ID-AGT FCM-C1q FCM-C1q ABO A/B-IgG A/B-IgG A/B-IgG A/B-IgG Ritux- 42

5 MHC 2016; 23 (1) imab IV. 結語 FCM-C1q A/B ABO 引用文献 1) Sawada T, Fuchinoue S, Kawase T, et al.: Preconditioning regimen consisting of anti-cd20 monoclonal antibody infusions, splenectomy and DFPP-enabled non-responders to undergo ABO-incompatible kidney transplantation. Clin Transplant (18): , ) Complementdependent cytotoxic crossmatch Flow cytometric crossmatch HLA 48: , ) Knight RC: Measuring IgG anti-a/b titres using dithiothreitol (DTT). Journal of Clinical Pathology (31): , ) Olson PR, Weiblen BJ, O Leary JJ, et al.: A simple technique for the inactivation of IgM antibodies using Dithiothreitol. Vox Sanguinis (30): , ) A/B 58: , ) Alexandre GP, De Bruyere M, Squifflet JP, et al.: Human ABOincompatible living donor renal homografts. Neth J Med (28): , ) Alexandre GP, Squifflet JP, De Bruye re M, et al.: ABOincompatible related and unrelated living donor renal allografts. Transplantation Proceedings (18): , ) Fuchinoue S, Ishii Y, Sawada T, et al.: The 5-Year Outcome of ABO-Incompatible Kidney Transplantation With Rituximab Induction. Transplantation (91): , ) Aikawa A, Kawamura T, Shishido S, et al.: ABO-incompatible living-donor pediatric kidney transplantation in Japan. Clinics (69): 22 27, ) Duncan AR, Winter G: The binding site for C1q on IgG. Nature 1988, 332, ) McVey J, Baker D, Parti R, et al.: Anti-A and anti-b titers in donor plasma, plasma pools, and immunoglobulin final products. Transfusion (55): S98 104, ) Grundbacher FJ: Genetics of anti-a and anti-b levels. Transfusion (16): 48 55, ) Baldwin WM 3rd, Pruitt SK, Brauer RB, et al.: Complement in organ transplantation. Contributions to inflammation, injury, and rejection. Transplantation (59): , ) A B 17 50: 350, ) ABO AMR 2: 62 67,

6 MHC 2016; 23 (1) Novel Flow Cytometry Method for Detecting Complement C1q Bound to Antiblood Group A/B-IgG Antibody in ABO-incompatible Renal Transplantation Tsutomu Ishizuka 1), Mineko Yasuo 1), Yuri Kobayashi 1), Hitomi Miura 1), Kotaro Kai 2), Kazuhiro Iwadoh 2), Toru Murakami 2), Kumiko Kitajima 2), Ichiro Nakajima 2) and Shohei Fuchinoue 2) 1) Division of Transplant Immunology, Central Clinical Laboratories, Tokyo Women s Medical University 2) Department of Surgery, Kidney Center, Tokyo Women s Medical University It has been widely believed that it is critical to remove as much anti A/B blood group antibody as possible before ABO incompatible renal transplantation (ABOI-RTx) for renal graft survival. We, however, report a rare case that implied a presence of hyperacute rejection in ABOI-RTx leading to graft loss, in a way, regardless of titers of blood group antibody before transplantation. We newly developed a method for measuring a complement-fixing ability of anti A/B blood group IgG antibody to elucidate an immunological mechanism of how anti A/B blood group antibody works in ABOI-RTx. Results of such measurements revealed that a high positive rate of complement-fixing ability of anti A/B IgG antibody was predictive of poor graft survival or graft loss. This method, therefore, might turn out to be an important laboratory test before ABOI-RTx. Key words: anti-a/b blood type antibody, complement-fixing ability of anti-a/b antibody, FCM-C1q, FCM-IgG, ABO incompatible renal transplantation

日本組織適合性学会誌第20巻2号

日本組織適合性学会誌第20巻2号 Major Histocompatibility Complex 2013; 20 (2): 121 129 平成 25 年度認定 HLA 検査技術者講習会テキスト HLA 1) 1) HLA HLA HLA PRA Panel reactive antibody DSA Donor specific alloantibody class subclass non-hla HLA キーワード :Human