We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.
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Method Article
Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET
* These authors contributed equally
In This Article
Summary
Abstract
The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.
Protocol
1. Donor Organ Recovery
- Set up the stereotactic microscope, record weight of the 8-10 week old rat (donor and recipient body weight should match).
- Anesthetize the donor rat (Lewis Brown Norway F1, LBN F1) using oxygen/isoflurane inhalation (isoflurane 4 %/2 L/min oxygen). Maintain anesthesia by lowering isoflurane to 2-2.5%.
- Place the anesthetized rat on the surgery pad. Fix the rat's extremities with adhesive tape onto the pad and apply ophthalmic ointment (Bepanthen, Bayer) to the rat's eyes.
- Shave and disinfect the abdomen of the rat and perform a ventral midline incision from the pubis to the caudal border of the liver. Ex....
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Results
Histology
During AR leukocytes, i.e. mainly T-lymphocytes are recruited into the transplant, whereas the severity of the rejection is reflected by the degree of inflammation. In the Periodic-Acid-Schiff (PAS) staining depicted here (Figure 1), the renal allograft shows significant histological signs of AR, namely glomerulitis, tubulitis, endothelialitis and graft infiltration (Figure 1, aTX POD4) (POD = postoperative day) while signs of rejection a.......
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Discussion
FDG-PET imaging is a new option for the diagnosis of acute rejection. Because of its non-invasive and specific nature, FDG-PET is advantages in comparison to classical diagnostics by core needle biopsy. In contrast to the limited sample size of a biopsy, FDG-PET analysis the whole graft. Moreover, one can apply it to patients on anticoagulant therapy, and one can perform PET measures repetitively e.g. to monitor treatment efficiency 4. In addition, we already have shown that two major different.......
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Disclosures
No conflicts of interest declared.
Acknowledgements
This work was supported by the Deutsche Forschungsgemeinschaft (SFB 656, Münster, Germany, projects C7 & C6) and the IZKF Münster (Core unit SmAP). The authors are grateful to Truc Van Le, Anne Kanzog, Ute Neugebauer, Wiebke Gottschlich and Roman Priebe for excellent technical assistance and to Daniel Burkert and Sven Fatum for producing radiotracers.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Equipment | |||
| Mathieu Needle Holder - 14 cm | Fine Science Tools | 12010-14 | |
| Castroviejo Micro Needle Holder - 9 cm | Fine Science Tools | 12061-01 | |
| Surgical Scissors - Sharp_Blunt | Fine Science Tools | 14001-12 | |
| Iris Scissors - ToughCut Straight 11.5 cm | Fine Science Tools | 14058-11 | |
| Student Vannas Spring Scissors | Fine Science Tools | 91500-09 | |
| Vannas Spring Scissors - 3 mm Blades | Fine Science Tools | 15000-00 | |
| Student Tissue Forceps - 1x2 Teeth 12 cm | Fine Science Tools | 91121-12 | |
| Dumont SS-45 Forceps - Inox Medical | Fine Science Tools | 11203-25 | |
| Micro-Serrefine Clip Applicator with Lock | Fine Science Tools | 18056-14 | |
| Micro-Serrefine 6 mm x 1 mm | Fine Science Tools | 18055-03 | |
| Micro-Serrefine 4 mm x 0.75 mm | Fine Science Tools | 18055-04 | |
| Reagent | |||
| Isoflurane (e.g. Forene 100% v/v) | Abott | ||
| cutane antiseptic (e.g. Octeniderm) | Schülke | ||
| Povidone Iodine (e.g. Betaisodona) | Mundipharma | ||
| ophthalmic ointment (e.g. Bepanthen) | Bayer | ||
| Buprenorphin (e.g. Temgesic) | RB Pharmaceuticals | ||
| HTK perfusion solution (e.g. CUSTODIOL HTK) | Dr. Franz Köhler Chemie | ||
| surgical thread Mersilene 0 | Ethicon | EH6665E | |
| surgical thread Mersilene 4-0 | Ethicon | EH6732H | |
| surgical thread Prolene 6-0 | Ethicon | 8697H | |
| surgical thread Ethilon 9-0 | Ethicon | 2809G | |
| surgical silk 5-0 | Vömel | 14739 | |
| Canula (e.g. Microlance 3, 27G ¾) | BD | 302200 |
References
- Wu, O., Levy, A. R., Briggs, A., Lewis, G., Jardine, A. Acute rejection and chronic nephropathy: a systematic review of the literature. Transplantation. 87, 1330-1339 (2009).
- Miller, B. F., Gonzales, E., Wilchins, L. J., Nathan, P. Kidney transplantation in the rat. Nature. 194
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