Removal of therapeutic anti-lymphocyte antibodies from human sera prior to anti-human leukocyte antibody testing

被引:16
作者
Bearden, CM
Book, BK
Sidner, RA
Pescovitz, MD
机构
[1] Indiana Univ, Dept Surg, Indianapolis, IN 46202 USA
[2] Indiana Univ, Dept Microbiol & Immunol, Indianapolis, IN 46204 USA
关键词
transplant; histocompatibility; crossmatch; antibodies; interference; PRA;
D O I
10.1016/j.jim.2005.03.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Both monoclonal (e.g. Orthoclone (OKT3 (R)), rituximab) and polyclonal (e.g. ATGAM (R), Thymoglobulin (R) (Thymo)) antilymphocyte Abs (ALAs) are used extensively in organ transplantation for immunosuppression induction, desensitization, and treatment of acute rejection. ALAs often interfere with post transplant immunologic monitoring. We describe a method that uses magnetic beads to selectively remove ALAs from patient serum. Rabbit anti-mouse Fc-specific (180 mu g), or rabbit anti-mouse Fab-specific (180 mu g), or rabbit anti-horse heavy and light chain-specific and rabbit anti-horse F(ab')(2) (200 mu g) (Jackson Immunoresearch) was adsorbed to 6.7 x 10(8) Dynabeads M-280 conjugated with sheep anti-rabbit IgG (Dynal Biotech). Fifty microliters of normal human serum (NHS) with 2 mu g/ml of OKT3 or 100 mu g/ml ATGAM, Thymo, or rituximab were incubated with conjugated beads for several incubations. NHS containing ALAs before and after treatment by the protocol were incubated with human lymphocytes and labeled with FITC-antibody to immunoglobulin of the species used to produce the particular ALA. Residual ALA was determined using flow cytometry. Average median channel for serum with or without ALA was 11.1 and 0.120, respectively for OKT3; 64.4 and 0.344 for ATGAM; 108.5 and 0.200 for Thymo; and 1022.5 and 11.4 for rituximab. Treatment lowered the median channel for serum with OKT3 to 0.103, 0.309 for ATGAM, 0.199 for Thymo, and 12.1 for rituximab. ALAs can be effectively removed from serum by the use of magnetic beads conjugated with Ab specific for ALA thereby permitting immunologic monitoring without interference. (c) 2005 Elsevier B.V All rights reserved.
引用
收藏
页码:192 / 199
页数:8
相关论文
共 26 条
[1]   A SIMPLE MICRO CYTOTOXICITY TEST [J].
AMOS, DB ;
BASHIR, H ;
BOYLE, W ;
MACQUEEN, M ;
TIILIKAINEN, A .
TRANSPLANTATION, 1969, 7 (03) :220-+
[2]   Anti-CD20 monoclonal antibody (rituximab) therapy for acute cardiac humoral rejection: A case report [J].
Aranda, JM ;
Scornik, JC ;
Normann, SJ ;
Lottenberg, R ;
Schofield, RS ;
Pauly, DF ;
Miles, M ;
Hill, JA ;
Sleasman, JW ;
Skoda-Smith, S .
TRANSPLANTATION, 2002, 73 (06) :907-910
[3]  
Barlow CN, 2001, TRANSPLANT INT, V14, P234, DOI 10.1007/s001470100314
[4]   Pronase treatment facilitates alloantibody flow cytometric and cytotoxic crossmatching in the presence of rituximab [J].
Bearden, CA ;
Agarwal, A ;
Book, BK ;
Sidner, RA ;
Gebel, HM ;
Bray, RA ;
Pescovitz, MD .
HUMAN IMMUNOLOGY, 2004, 65 (08) :803-809
[5]  
BonnefoyBerard N, 1996, J HEART LUNG TRANSPL, V15, P435
[6]   ANTIBODIES AGAINST FUNCTIONAL LEUKOCYTE SURFACE MOLECULES IN POLYCLONAL ANTILYMPHOCYTE AND ANTITHYMOCYTE GLOBULINS [J].
BONNEFOYBERARD, N ;
VINCENT, C ;
REVILLARD, JP .
TRANSPLANTATION, 1991, 51 (03) :669-673
[7]  
BONNEFOYBERARD N, 1992, IMMUNOLOGY, V77, P61
[8]  
BRAY RA, 1994, METHOD CELL BIOL, V41, P103
[9]   A randomized, double-blinded comparison of thymoglobulin versus Atgam for induction immunosuppressive therapy in adult renal transplant recipients [J].
Brennan, DC ;
Flavin, K ;
Lowell, JA ;
Howard, TK ;
Shenoy, S ;
Burgess, S ;
Dolan, S ;
Kano, JM ;
Mahon, M ;
Schnitzler, MA ;
Woodward, R ;
Irish, W ;
Singer, GG .
TRANSPLANTATION, 1999, 67 (07) :1011-1018
[10]  
CARRIER M, 1993, CAN J CARDIOL, V9, P171