Evaluation of a gene expression biomarker to identify operationally tolerant liver transplant recipients: the LITMUS trial

Andrzej Chruscinski, Vanessa Rojas-Luengas, Sajad Moshkelgosha, Assaf Issachar, Jane Luo, Handy Yowanto, Leslie Lilly, Robert Smith, Eberhard Renner, Jianhua Zhang, Maor Epstein, David Grant, Caitriona M. Mcevoy, Ana Konvalinka, Atul Humar, Oyedele Adeyi, Sandra Fischer, Felix H. Volmer, Richard Taubert, Elmar JaeckelStephen Juvet, Nazia Selzner, Gary A. Levy

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


LITMUS was a single-centre, Phase 2a study designed to investigate whether the gene biomarker FGL2/IFNG previously reported for the identification of tolerance in murine models could identify operationally tolerant liver transplant recipients. Multiplex RT-PCR was used to amplify eight immunoregulatory genes in peripheral blood mononuclear cells (PBMC) from 69 adult liver transplant recipients. Patients with PBMC FGL2/IFNG ≥ 1 and a normal liver biopsy underwent immunosuppression (IS) withdrawal. The primary end point was the development of operational tolerance. Secondary end points included correlation of tolerance with allograft gene expression and immune cell markers. Twenty-eight of 69 patients (38%) were positive for the PBMC tolerance biomarker and 23 proceeded to IS withdrawal. Nine of the 23 patients had abnormal baseline liver biopsies and were excluded. Of the 14 patients with normal biopsies, eight (57%) have achieved operational tolerance and are off IS (range 12-57 months). Additional studies revealed that all of the tolerant patients and only one non-tolerant patient had a liver gene ratio of FOXP3/IFNG ≥ 1 prior to IS withdrawal. Increased CD4+ T regulatory T cells were detected both in PBMC and livers of tolerant patients following IS withdrawal. Higher expression of SELE (gene for E-selectin) and lower expression of genes associated with inflammatory responses (GZMB, CIITA, UBD, LSP1, and CXCL9) were observed in the pre-withdrawal liver biopsies of tolerant patients by RNA sequencing. These results suggest that measurement of PBMC FGL2/IFNG may enrich for the identification of operationally tolerant liver transplant patients, especially when combined with intragraft measurement of FOXP3/IFNG.

Original languageEnglish (US)
Pages (from-to)123-139
Number of pages17
JournalClinical and Experimental Immunology
Issue number1
StatePublished - Jan 2022
Externally publishedYes

Bibliographical note

Funding Information:
LITMUS was supported by a grant from the Heart and Stroke Foundation of Canada (G-17-0018658) and a generous donation from Dr. Ellen Bialystok and Dr. Franklin Bialystok.

Publisher Copyright:
© 2021 The Author(s).


  • biomarker
  • gene expression
  • immunosuppression withdrawal
  • liver transplantation
  • tolerance
  • Graft Rejection/diagnosis
  • Gene Expression
  • Immunosuppressive Agents
  • Humans
  • Biomarkers/metabolism
  • Fibrinogen
  • Immune Tolerance/genetics
  • Adult
  • Transplantation Tolerance/genetics
  • Leukocytes, Mononuclear/metabolism
  • Liver Transplantation/methods

PubMed: MeSH publication types

  • Clinical Trial, Phase II
  • Journal Article
  • Research Support, Non-U.S. Gov't


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