title: Towards functional transplant donor matching by measurement of granzyme A and granzyme B production levels.
abstract: Graft-versus-host disease (GvHD) can be a major complication after allogeneic stem cell transplantation (SCT) especially when donor and recipient are unrelated. The latter serious complication, together with the growing number of available unrelated stem cell donors, demand a simple in vitro assay for functional stem cell donor selection. Activated donor cytotoxic T lymphocytes (CTLs) and natural killer cells produce granzymes (Gr) that are involved in the pathogenesis of GvHD. We measured granzymes A and B (GrA and GrB) production levels in the supernatants of 96 h pretransplant mixed lymphocyte cultures (MLC) of 26 sibling and 31 unrelated patient/donor pairs by enzyme-linked immunosorbent assay (ELISA). In detail, the GrA and GrB production levels from a selected cohort of 37 potential patient/donor pairs were correlated with relative responses (RR) of MLC and with human leukocyte antigen (HLA) class II mismatches and with the development of acute GvHD in a second, consecutive cohort of 20 sibling SCT recipients. In vitro measurement of GrA and GrB production levels significantly correlated with the RR of pretransplant MLC (r=0.492, p< or =0.01 and r=0.853, p< or =0.01, respectively) and increased with the number of HLA class II mismatches between patient and donor. Pretransplant GrA production levels were significantly associated with the in vivo development of acute GvHD grades II-IV in patients transplanted with an HLA-identical sibling donor (p< or =0.001). In conclusion, in vitro GrA and GrB production levels can be measured by a quantitative and sensitive ELISA. This novel and simple method may be used for functional selection of unrelated stem cell donors and for the identification of patients who are at risk for acute GvHD grades II-IV.
authors.affiliation: Laboratory for Tumor and Immunobiology, Department of Hematology and Oncology, Innsbruck Medical University, Anichstrasse 35, 6020 Innsbruck, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Kircher
authors.forename: C Erik
authors.initials: CE
authors.lastname: Hack
authors.forename: Anne M
authors.initials: AM
authors.lastname: Dickinson
authors.forename: Xiao N
authors.initials: XN
authors.lastname: Wang
authors.forename: Machteld
authors.initials: M
authors.lastname: Oudshoorn
authors.forename: Annemarie
authors.initials: A
authors.lastname: Sachs
authors.forename: Angela
authors.initials: A
authors.lastname: Wolbink
authors.forename: Dietger
authors.initials: D
authors.lastname: Niederwieser
authors.forename: Günther J
authors.initials: GJ
authors.lastname: Eibl
authors.forename: Hans C
authors.initials: HC
authors.lastname: van Houwelingen
authors.forename: Els
authors.initials: E
authors.lastname: Goulmy
mesh_terms: D046148:Donor Selection; D006086:Graft vs Host Disease / Q000517:prevention & control*; D053804:Granzymes; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D012697:Serine Endopeptidases / Q000276:immunology / Q000378:metabolism*; D014019:Tissue Donors
chemical_list: D000949:Histocompatibility Antigens Class II; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human