PMID- 24510725
OWN - NLM
STAT- MEDLINE
DCOM- 20140930
LR  - 20200108
IS  - 1934-9300 (Electronic)
IS  - 1934-9297 (Linking)
VI  - 66
DP  - 2013 Oct 9
TI  - Flow cytometry-based cytotoxicity and antibody binding assay.
PG  - 6.34.1-6.34.11
LID - 10.1002/0471142956.cy0634s66 [doi]
AB  - Human leukocyte antigen (HLA) antibodies with the ability to activate complement 
      are associated with an increased risk of early antibody-mediated graft rejection 
      in kidney transplantation (KTx). Detection of these potentially harmful 
      complement-fixing HLA antibodies is commonly performed via the 
      complement-dependent cytotoxicity (CDC) assay according to protocols that were 
      developed as early as 40 years ago. The read-out for this assay is based on 
      manual scoring by visual inspection of cells under a fluorescence microscope. CDC 
      is often used in combination with the flow cytometry-based lymphocyte crossmatch 
      assay (FCXM), which, with high sensitivity, detects HLA antibody binding. Here we 
      describe a new approach wherein both cytotoxicity and antibody binding can be 
      simultaneously assessed with flow cytometry. Two strategies are described, using 
      either magnetic bead-enriched T and B lymphocytes or bulk peripheral blood 
      mononuclear cells (PBMC) as donor target cells.
CI  - Copyright (c) 2013 John Wiley & Sons, Inc.
FAU - Alheim, Mats
AU  - Alheim M
AD  - Department of Laboratory Medicine, Division of Clinical Immunology and 
      Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden.
LA  - eng
PT  - Journal Article
DEP - 20131009
PL  - United States
TA  - Curr Protoc Cytom
JT  - Current protocols in cytometry
JID - 100899351
RN  - 0 (Antibodies)
SB  - IM
MH  - Antibodies/*metabolism
MH  - B-Lymphocytes/immunology
MH  - Blood Grouping and Crossmatching
MH  - Cytotoxicity Tests, Immunologic/*methods
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - Microspheres
MH  - T-Lymphocytes/immunology
MH  - Tissue Donors
OTO - NOTNLM
OT  - HLA antibodies
OT  - complement dependent cytotoxicity
OT  - crossmatching
OT  - flow cytometry
OT  - kidney transplantation
EDAT- 2014/02/11 06:00
MHDA- 2014/10/01 06:00
CRDT- 2014/02/11 06:00
PHST- 2014/02/11 06:00 [entrez]
PHST- 2014/02/11 06:00 [pubmed]
PHST- 2014/10/01 06:00 [medline]
AID - 10.1002/0471142956.cy0634s66 [doi]
PST - epublish
SO  - Curr Protoc Cytom. 2013 Oct 9;66:6.34.1-6.34.11. doi: 
      10.1002/0471142956.cy0634s66.