The use of granzyme A as a marker of heart transplant rejection in cyclosporine or anti-CD4 monoclonal antibody-treated rats.
Granzyme A is a serine protease expressed by populations of human and mouse natural killer cells and activated CD4+ and CD8+ cytotoxic lymphocytes; its expression marks a subset of inflammatory cells in allografts, autoimmune diabetes, and a number of other inflammatory lesions. In order to describe more completely the correlation between granzyme A expression and the presence of in vivo cytolytic effects, we grafted allogeneic rat hearts with vascular anastomoses in a heterotopic location, and treated the hosts with either cyclosporine, anti-CD4 monoclonal antibody (MRC OX38), or no therapy. The grafts were evaluated by palpation for cardiac functions, by immunohistochemistry for CD4/CD8 expression, by hematoxylin-and-eosin staining for inflammatory infiltration, and by in situ hybridization for granzyme A expression. The appearance of granzyme A+ cells in untreated allografts preceded both functional and standard histopathological and immunohistochemical evidence of graft rejection by two days. In donor-recipient combinations where cyclosporine and anti-CD4 treatments allowed indefinite allograft survival, the allografts showed minimal numbers of granzyme A+ cells, whether cellular infiltrates developed or not. The number of granzyme A+ cells present in the cardiac allografts in treated and untreated animals correlated with either current or impending episodes of rejection. The early time course of granzyme A expression suggests that it can be used as an early and reliable marker of graft rejection.
Chen
R H
RH
Howard Hughes Medical Institute, Stanford University School of Medicine, California 94305.
Ivens
K W
KW
Alpert
S
S
Billingham
M E
ME
Fathman
C G
CG
Flavin
T F
TF
Shizuru
J A
JA
Starnes
V A
VA
Weissman
I L
IL
Griffiths
G M
GM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D015704:CD4 Antigens / Q000276:immunology; D016516:CD4-CD8 Ratio; D016572:Cyclosporine / Q000494:pharmacology; D006084:Graft Rejection / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology; D006085:Graft Survival / Q000187:drug effects / Q000276:immunology; D053804:Granzymes; D016027:Heart Transplantation / Q000276:immunology*; D006648:Histocompatibility; D008297:Male; D051381:Rats; D011912:Rats, Inbred ACI; D011917:Rats, Inbred Lew; D012697:Serine Endopeptidases / Q000032:analysis*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014184:Transplantation, Homologous; D014185:Transplantation, Isogeneic
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D015704:CD4 Antigens; D016572:Cyclosporine; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human