Noninvasive Imaging of Acute Renal Allograft Rejection by Ultrasound Detection of Microbubbles Targeted to T-lymphocytes in Rats.
OBJECTIVE
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.


METHODS
15 minutes after tail vein injection of 30 × 10(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, 45 min. warm ischemia), and rats subjected to acute cyclosporin A toxicity (CSA) (cyclosporine 50 mg/kg BW for 2 days i. p.) served as controls.


RESULTS
Accumulation of human T-lymphocytes was clearly detected by antibody-mediated sonography und was significantly increased in allografts undergoing AR (5.41 ± 1.32 A. U.) when compared to native control kidneys (0.70 ± 0.08 A. U.). CD3 signal intensity was low in native kidneys, sTX (0.99 ± 0.30 A. U.), CSA (0.10 ± 0.02 A. U.) and kidneys with IRI (0.46 ± 0.29 A. U.). Quantification of the ultrasound signal correlated significantly with the T-cell numbers obtained by immunohistochemical analysis (R2 = 0.57).


CONCLUSIONS
Contrast-enhanced sonography using CD3-antibodies is an option for quick and highly specific assessment of AR in a rat model of renal transplantation.
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
A
A
Grabner
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
D
D
Kentrup
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
M
M
Mühlmeister
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
H
H
Pawelski
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
C
C
Biermann
Bracco Suisse SA, Bracco Suisse SA, Geneva, Italy.
T
T
Bettinger
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
H
H
Pavenstädt
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
E
E
Schlatter
Department of Nuclear Medicine, Technical University Munich, Germany.
K
K
Tiemann
Department of Medicine D, Experimental Nephrology, University Hospital Münster, Germany.
S
S
Reuter
D000208:Acute Disease; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D004195:Disease Models, Animal; D006084:Graft Rejection / Q000000981:diagnostic imaging* / Q000473:pathology; D007668:Kidney / Q000000981:diagnostic imaging / Q000473:pathology; D016030:Kidney Transplantation; D008297:Male; D045423:Microbubbles; D057054:Molecular Imaging; D051381:Rats; D011917:Rats, Inbred Lew; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology; D014463:Ultrasonography
D000906:Antibodies; D017252:CD3 Complex