title: Protein microarrays identify antibodies to protein kinase Czeta that are associated with a greater risk of allograft loss in pediatric renal transplant recipients.
abstract: Antibodies to human leukocyte antigens (HLAs) are a risk factor for acute renal allograft rejection and loss. The role of non-HLAs and their significance to allograft rejection have gained recent attention. Here, we applied protein microarray technology, with the capacity to simultaneously identify 5056 potential antigen targets, to assess non-HLA antibody formation in 15 pediatric renal transplant recipients during allograft rejection. Comparison of the pre- and post-transplant serum identified de novo antibodies to 229 non-HLA targets, 36 of which were present in multiple patients at allograft rejection. On the basis of its reactivity, protein kinase Czeta (PKCzeta) was selected for confirmatory testing and clinical study. Immunohistochemical analysis found PKCzeta both within the renal tissue and infiltrating lymphocytes at rejection. Patients who had an elevated anti-PKCzeta titer developed rejection, which was significantly more likely to result in graft loss. The absence of C4d deposition in patients with high anti-PKCzeta titers suggests that it is a marker of severe allograft injury rather than itself being pathogenic. Presumably, critical renal injury and inflammation associated with this rejection subtype lead to the immunological exposure of PKCzeta with resultant antibody formation. Prospective assessment of serum anti-PKCzeta levels at allograft rejection will be needed to confirm these results.
authors.affiliation: Division of Nephrology, Department of Pediatrics, Stanford University Medical Center, Stanford, CA 94305, USA. suthersm@stanford.edu
authors.forename: Scott M
authors.initials: SM
authors.lastname: Sutherland
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Tara K
authors.initials: TK
authors.lastname: Sigdel
authors.forename: Persis P
authors.initials: PP
authors.lastname: Wadia
authors.forename: David B
authors.initials: DB
authors.lastname: Miklos
authors.forename: Atul J
authors.initials: AJ
authors.lastname: Butte
authors.forename: Minnie M
authors.initials: MM
authors.lastname: Sarwal
mesh_terms: D000293:Adolescent; D002648:Child; D015935:Complement C4b / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006084:Graft Rejection / Q000201:enzymology / Q000209:etiology* / Q000276:immunology* / Q000473:pathology; D006085:Graft Survival / Q000276:immunology; D006680:HLA Antigens; D006801:Humans; D007150:Immunohistochemistry; D007518:Isoantibodies / Q000097:blood*; D007519:Isoantigens; D007668:Kidney / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D016030:Kidney Transplantation / Q000009:adverse effects* / Q000276:immunology* / Q000473:pathology; D008297:Male; D010446:Peptide Fragments / Q000378:metabolism; D040081:Protein Array Analysis; D011493:Protein Kinase C / Q000276:immunology* / Q000378:metabolism; D012307:Risk Factors; D014184:Transplantation, Homologous; D000099221:Protein Kinase C zeta
chemical_list: D015935:Complement C4b; D006680:HLA Antigens; D007518:Isoantibodies; D007519:Isoantigens; D010446:Peptide Fragments; D011493:Protein Kinase C; D000099221:Protein Kinase C zeta; C032261:complement C4d