ABSTRACT
Noninvasive methods to diagnose and differentiate acute cellular rejection from acute tubular necrosis or acute calcineurin inhibitor toxicity are still missing. Because T lymphocytes play a decisive role in early states of rejection, we investigated the suitability and feasibility of antibody-mediated contrast-enhanced ultrasound by using microbubbles targeted to CD3(+) , CD4(+) , or CD8(+) T cells in different models of renal disease. In an established rat renal transplantation model, CD3-mediated ultrasound allows the detection of acute rejection as early as on postoperative day 2. Ultrasound signal intensities increased with the severity of inflammation. Further, an early response to therapy could be monitored by using contrast-enhanced sonography. Notably, acute tubular necrosis occurring after ischemia-reperfusion injury as well as acute calcineurin inhibitor toxicity could easily be differentiated. Finally, the quantified ultrasound signal correlated significantly with the number of infiltrating T cells obtained by histology and with CD3 mRNA levels, as well as with chemokine CXCL9, CXCL11, and CCL19 mRNA but not with KIM-1 mRNA expression, thereby representing the severity of graft inflammation but not the degree of kidney injury. In summary, we demonstrate that antibody-mediated contrast-enhanced ultrasound targeting T lymphocytes could be a promising tool for an easy and reproducible assessment of acute rejection after renal transplantation.
Subject(s)
CD3 Complex/immunology , Graft Rejection/diagnosis , Kidney Transplantation/adverse effects , Molecular Imaging/methods , Reperfusion Injury/complications , T-Lymphocytes/immunology , Ultrasonography/methods , Acute Disease , Animals , Calcineurin Inhibitors/toxicity , Contrast Media/metabolism , Graft Rejection/diagnostic imaging , Graft Rejection/etiology , Isoantibodies/toxicity , Kidney Tubular Necrosis, Acute/diagnosis , Kidney Tubular Necrosis, Acute/diagnostic imaging , Kidney Tubular Necrosis, Acute/etiology , Male , Microbubbles , Rats , Rats, Inbred BN , Rats, Inbred Lew , Reperfusion Injury/surgery , Transplantation, HomologousABSTRACT
PURPOSE: We propose CD3-antibody-mediated contrast-enhanced ultrasonography using human T-lymphocytes for image-based diagnosis of acute allograft rejection (AR) established in a rat renal transplantation model. MATERIALS AND METHODS: 15 minutes after tail vein injection of 30â×â10(6) human T-lymphocytes, contrast media/microbubbles conjugated with an anti-human CD3 antibody was applied to uni-nephrectomized 10-week-old allogeneically transplanted male rats (Lewis-Brown Norway (LBN) to Lewis, aTX) and ultrasound was performed to investigate the transplanted kidney as well as the native kidney. In vivo results were confirmed via immunohistochemical stainings of CD3 after post mortem dissection. Syngeneically transplanted rats (LBN to LBN, sTX), rats with ischemia/reperfusion injury (IRI, 45âmin. warm ischemia), and rats subjected to acute cyclosporin A toxicity (CSA) (cyclosporine 50âmg/kg BW for 2 days i.âp.) served as controls. RESULTS: Accumulation of human T-lymphocytes was clearly detected by antibody-mediated sonography und was significantly increased in allografts undergoing AR (5.41â±â1.32 A.âU.) when compared to native control kidneys (0.70â±â0.08 A.âU.). CD3 signal intensity was low in native kidneys, sTX (0.99â±â0.30 A.âU.), CSA (0.10â±â0.02 A.âU.) and kidneys with IRI (0.46â±â0.29 A.âU.). Quantification of the ultrasound signal correlated significantly with the T-cell numbers obtained by immunohistochemical analysis (R2â=â0.57). CONCLUSION: Contrast-enhanced sonography using CD3-antibodies is an option for quick and highly specific assessment of AR in a rat model of renal transplantation.
