"FITC-labelled GzmB substrate peptides with internal quencher ((5-FAM)aIEFDSGK(CPQ2)kkc) were synthesized by CPC Scientific and used for all in vitro activity assays."
Abstract
The early detection of the onset of transplant rejection is critical for the long-term survival of patients. The diagnostic gold-standard for detecting transplant rejection involves a core biopsy, which is invasive, has limited predictive power, and carries a morbidity risk. Here, we show that nanoparticles conjugated with a peptide substrate specific for the serine protease granzyme B, which is produced by recipient T cells during the onset of acute cellular rejection, can serve as a non-invasive biomarker of early rejection. Upon systemic administration in mouse models of skin-graft rejection, these nanosensors preferentially accumulate in allograft tissue, where they are cleaved by granzyme B, releasing a fluorescent reporter that filters into the recipient’s urine. Urinalysis then discriminates the onset of rejection with high sensitivity and specificity before features of rejection are apparent in grafted tissues. Moreover, in mice treated with subtherapeutic levels of immunosuppressive drugs, the reporter signals in urine can be detected before graft failure. This method may enable routine monitoring of allograft status without the need for biopsies.
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