title: A rapid test to monitor alloreactive responses in whole blood using real-time polymerase chain reaction.
abstract: Rapid, simple, and reliable assays to monitor allogeneic responses are essential for the safe development of novel protocols of tailored immunosuppression. Herein, we describe a real-time polymerase chain reaction method based on interleukin-2 and interferon-gamma mRNA quantification upon stimulation of whole blood with allogeneic T cell-depleted peripheral blood mononuclear cells. The technique requires only small blood volumes and results can be obtained within 48 hours. Data obtained in a liver transplant patient receiving a tolerance induction protocol based on the infusion of donor-type hematopoietic stem cells suggest that this rapid whole blood mixed lymphocyte reaction assay could be valuable for the monitoring of patients undergoing solid organ or hematopoietic stem cell transplantation.
authors.affiliation: Department of Immunology-Hematology-Transfusion, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhou
authors.forename: Michel
authors.initials: M
authors.lastname: Toungouz
authors.forename: Vincent
authors.initials: V
authors.lastname: Donckier
authors.forename: Marc
authors.initials: M
authors.lastname: Andrien
authors.forename: Roberto
authors.initials: R
authors.lastname: Troisi
authors.forename: Bernard
authors.initials: B
authors.lastname: de Hemptinne
authors.forename: Alain
authors.initials: A
authors.lastname: Le Moine
authors.forename: Etienne
authors.initials: E
authors.lastname: Dupont
authors.forename: Michel
authors.initials: M
authors.lastname: Goldman
authors.forename: Patrick
authors.initials: P
authors.lastname: Stordeur
mesh_terms: D000917:Antibody Formation; D017252:CD3 Complex / Q000096:biosynthesis; D016207:Cytokines / Q000096:biosynthesis*; D005434:Flow Cytometry; D006684:HLA-DR Antigens; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006650:Histocompatibility Testing; D006801:Humans; D007158:Immunologic Techniques; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D016031:Liver Transplantation; D007959:Lymphocyte Culture Test, Mixed; D010455:Peptides / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D013997:Time Factors; D018183:Transplantation Chimera; D023001:Transplantation Tolerance; D014184:Transplantation, Homologous / Q000379:methods*
chemical_list: D017252:CD3 Complex; D016207:Cytokines; D006684:HLA-DR Antigens; D007376:Interleukin-2; D010455:Peptides; D012333:RNA, Messenger; D007371:Interferon-gamma