Immunohistochemistry staining of C4d to diagnose antibody-mediated rejection in cardiac transplantation.
BACKGROUND
Antibody-mediated rejection (AMR) is associated with poorer outcomes in cardiac transplantation. The clinical diagnosis of AMR has been confirmed by immunofluorescence for C4d on fresh-frozen cardiac tissue. Immunohistochemistry (IHC) has been suggested as a more practical diagnostic tool because it can be performed on routine paraffin-embedded tissue. There are few published data about hemodynamics and C4d staining. We prospectively performed C4d staining on endomyocardial biopsies (EMBs) and present the pattern of tissue staining and its correlation with intracardiac hemodynamics.


METHODS
EMBs were evaluated by IHC for C4d staining and graded for cellular rejection using ISHLT criteria on hematoxylin-and-eosin-stained sections. Hemodynamic measurements were taken concurrently. Staining for C4d was described as absent, present with serum staining, or present with only tissue staining. The pattern of tissue staining was categorized by location of staining and correlated with intracardiac hemodynamics. Patient demographics, cytomegalovirus status, panel-reactive antibody levels and hemodynamics were analyzed by analysis of variance and chi-square statistics.


RESULTS
Of the 400 EMBs, 50 had no C4d staining, 330 had tissue and serum staining, whereas 20 had only tissue staining. Forty EMBs had endothelial staining, including 35 with serum and 5 with isolated tissue staining. Endothelial staining correlated with higher intracardiac pressures.


CONCLUSIONS
IHC staining for C4d has been suggested for the diagnosis of AMR. Our data suggest there is a high rate of background C4d staining, but endothelial staining correlates with poorer hemodynamics. Methods for IHC staining and interpretation need to be standardized for widespread use and clinical studies.
Fedson
Savitri E
SE
Department of Medicine, University of Chicago Medical Center, Chicago, Illinois 60637, USA. sfedson@medicine.bsd.uchicago.edu
Daniel
Silver S
SS
Husain
Aliya N
AN
D000328:Adult; D000906:Antibodies / Q000276:immunology*; D015935:Complement C4b / Q000378:metabolism*; D003326:Coronary Circulation; D004699:Endocardium / Q000378:metabolism; D004727:Endothelium / Q000378:metabolism; D005260:Female; D005500:Follow-Up Studies; D006084:Graft Rejection / Q000175:diagnosis* / Q000276:immunology* / Q000401:mortality / Q000503:physiopathology; D016027:Heart Transplantation; D006439:Hemodynamics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009206:Myocardium / Q000378:metabolism; D010446:Peptide Fragments / Q000097:blood / Q000378:metabolism*; D011446:Prospective Studies; D013194:Staining and Labeling; D014018:Tissue Distribution
D000906:Antibodies; D010446:Peptide Fragments; D015935:Complement C4b; C032261:complement C4d