title: A Three-Gene Assay for Monitoring Immune Quiescence in Kidney Transplantation.
abstract: Organ transplant recipients face life-long immunosuppression and consequently are at high risk of comorbidities. Occasionally, kidney transplant recipients develop a state of targeted immune quiescence (operational tolerance) against an HLA-mismatched graft, allowing them to withdraw all immunosuppression and retain stable graft function while resuming immune responses to third-party antigens. Methods to better understand and monitor this state of alloimmune quiescence by transcriptional profiling may reveal a gene signature that identifies patients for whom immunosuppression could be titrated to reduce patient and graft morbidities. Therefore, we investigated 571 unique peripheral blood samples from 348 HLA-mismatched renal transplant recipients and 101 nontransplant controls in a four-stage study including microarray, quantitative PCR, and flow cytometry analyses. We report a refined and highly validated (area under the curve, 0.95; 95% confidence interval, 0.92 to 0.97) peripheral blood three-gene assay (KLF6, BNC2, CYP1B1) to detect the state of operational tolerance by quantitative PCR. The frequency of predicted alloimmune quiescence in stable renal transplant patients receiving long-term immunosuppression (n=150) was 7.3% by the three-gene assay. Targeted cell sorting of peripheral blood from operationally tolerant patients showed a significant shift in the ratio of circulating monocyte-derived dendritic cells with significantly different expression of the genes constituting the three-gene assay. Our results suggest that incorporation of patient screening by specific cellular and gene expression assays may support the safety of drug minimization trials and protocols.
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California;
authors.forename: Silke
authors.initials: S
authors.lastname: Roedder
authors.affiliation: Department of Biostatistics, Mount Sinai School of Medicine, New York, New York;
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Pathology, Stanford University, Palo Alto, California;
authors.forename: Michael N
authors.initials: MN
authors.lastname: Alonso
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California;
authors.forename: Szu-Chuan
authors.initials: SC
authors.lastname: Hsieh
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California;
authors.forename: Minh Thien
authors.initials: MT
authors.lastname: Vu
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California;
authors.forename: Hong
authors.initials: H
authors.lastname: Dai
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California;
authors.forename: Tara K
authors.initials: TK
authors.lastname: Sigdel
authors.affiliation: Department of Surgery, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico; and.
authors.forename: Ian
authors.initials: I
authors.lastname: Bostock
authors.affiliation: Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
authors.forename: Camila
authors.initials: C
authors.lastname: Macedo
authors.affiliation: Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
authors.forename: Diana
authors.initials: D
authors.lastname: Metes
authors.affiliation: Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
authors.forename: Adrianna
authors.initials: A
authors.lastname: Zeevi
authors.affiliation: Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
authors.forename: Ron
authors.initials: R
authors.lastname: Shapiro
authors.affiliation: Department of Pathology, Stanford University, Palo Alto, California;
authors.forename: Oscar
authors.initials: O
authors.lastname: Salvatierra
authors.affiliation: Department of Pathology, Stanford University, Palo Alto, California;
authors.forename: John
authors.initials: J
authors.lastname: Scandling
authors.affiliation: Department of Surgery, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico; and.
authors.forename: Josefina
authors.initials: J
authors.lastname: Alberu
authors.affiliation: Department of Pathology, Stanford University, Palo Alto, California;
authors.forename: Edgar
authors.initials: E
authors.lastname: Engleman
authors.affiliation: Department of Surgery, Division of Transplant Surgery, University of California San Francisco, San Francisco, California; minnie.sarwal@ucsf.edu.
authors.forename: Minnie M
authors.initials: MM
authors.lastname: Sarwal
mesh_terms: D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000097:blood*; D001772:Blood Cell Count; D039521:CD11c Antigen / Q000378:metabolism; D016022:Case-Control Studies; D002648:Child; D065633:Cytochrome P-450 CYP1B1 / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007165:Immunosuppression Therapy; D016030:Kidney Transplantation; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D014181:Transplantation Immunology / Q000235:genetics*; D055815:Young Adult
chemical_list: C484527:BNC2 protein, human; D015415:Biomarkers; D039521:CD11c Antigen; D004268:DNA-Binding Proteins; C105465:KLF6 protein, human; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors; D011518:Proto-Oncogene Proteins; C585496:CYP1B1 protein, human; D065633:Cytochrome P-450 CYP1B1
keywords: immunosuppression; kidney transplantation; profiling; tolerance; transcriptional