Name: non-invasive imaging of acute allograft rejection after rat renal transplantation using 18f-fdg pet
Uploaded: 2013-04-28T14:00:00
Description: Watch this Scientific Journal Video about Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET at JoVE.com

This work was supported by the Deutsche Forschungsgemeinschaft (SFB 656, M&uuml;nster, Germany, projects C7 &amp; C6) and the IZKF M&uuml;nster (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.

1. Donor Organ Recovery
<ol>
 <li>Set up the stereotactic microscope, record weight of the 8-10 week old rat (donor and recipient body weight should match).</li>
 <li>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%.</li>
 <li>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 ...

<ol>
	<li>Wu, O., Levy, A. R., Briggs, A., Lewis, G., Jardine, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+and+chronic+nephropathy:+a+systematic+review+of+the+literature.">Acute rejection and chronic nephropathy: a systematic review of the literature.</a> Transplantation. 87, 1330-1339 (2009).</li><li>Miller, B. F., Gonzales, E., Wilchins, L. J., Nathan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplantation+in+the+rat.">Kidney transplantation in the rat.</a> Nature. 194, 309-310 (1962).</li><li>Reuter, S., Schn&#246;ckel, U., Schr&#246;ter, R., Schober, O., Pavenst&#228;dt, H., Sch&#228;fers, M., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+imaging+of+acute+renal+allograft+rejection+in+rats+using+small+animal+F-FDG-PET.">Non-invasive imaging of acute renal allograft rejection in rats using small animal F-FDG-PET.</a> PLoS. One. 4, e5296 (2009).</li><li>Reuter, S., Schn&#246;ckel, U., Edemir, B., Schr&#246;ter, R., Kentrup, D., Pavenst&#228;dt, H., Schober, O., Schlatter, E., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+of+noninvasive+serial+assessment+of+acute+renal+allograft+rejection+by+18F-FDG+PET+to+monitor+treatment+efficiency.">Potential of noninvasive serial assessment of acute renal allograft rejection by 18F-FDG PET to monitor treatment efficiency.</a> J. Nucl. Med. 51, 1644-1652 (2010).</li><li>Schn&#246;ckel, U., Reuter, S., Stegger, L., Schlatter, E., Sch&#228;fers, K. P., Hermann, S., Schober, O., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+18F-fluoride+small+animal+PET+to+noninvasively+assess+renal+function+in+rats.">Dynamic 18F-fluoride small animal PET to noninvasively assess renal function in rats.</a> Eur. J. Nucl. Med. Mol. Imaging. 35, 2267-2274 (2008).</li><li>Grabner, A., Schn&#246;ckel, U., Kentrup, D., Sch&#228;fers, M., Reuter, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+for+Non-Invasive+Molecular+Imaging+of+Acute+Allograft+Rejection+by+Gamma+Scintigraphy+and+Positron+Emission+Tomography.">Strategies for Non-Invasive Molecular Imaging of Acute Allograft Rejection by Gamma Scintigraphy and Positron Emission Tomography.</a> Curr. Radiopharm. 4, 10-23 (2011).</li><li>Racusen, L. C., Solez, K., Colvin, R. B., Bonsib, S. M., Castro, M. C., Cavallo, T., Croker, B. P., Demetris, A. J., Drachenberg, C. B., Fogo, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Banff+97+working+classification+of+renal+allograft+pathology.">The Banff 97 working classification of renal allograft pathology.</a> Kidney Int. 55, 713-723 (1999).</li><li>Edemir, B., Reuter, S., Borgulya, R., Schr&#246;ter, R., Neugebauer, U., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+modulates+gene+expression+in+the+collecting+duct.">Acute rejection modulates gene expression in the collecting duct.</a> J. Am. Soc. Nephrol. 19, 538-546 (2008).</li><li>Velic, A., Gabri&#235;ls, G., Hirsch, J. R., Schr&#246;ter, R., Edemir, B., Paasche, S., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+after+rat+renal+transplantation+leads+to+downregulation+of+Na++and+water+channels+in+the+collecting+duct.">Acute rejection after rat renal transplantation leads to downregulation of Na+ and water channels in the collecting duct.</a> Am. J. Transplant. 5, 1276-1285 (2005).</li><li>Reuter, S., Velic, A., Edemir, B., Schr&#246;ter, R., Pavenst&#228;dt, H., Gabri&#235;ls, G., Bleich, M., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+role+of+NHE-3+inhibition+in+rat+renal+transplantation+undergoing+acute+rejection.">Protective role of NHE-3 inhibition in rat renal transplantation undergoing acute rejection.</a> Pflugers Arch. 456, 1075-1084 (2008).</li></ol>

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...

<table border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Mathieu Needle Holder - 14 cm</td>
 <td>Fine Science Tools</td>
 <td>12010-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Castroviejo Micro Needle Holder - 9 cm</td>
 <td>Fine Science Tools</td>
 <td>12061-01</td>
 <td></td>
 </tr>
 <tr>
 <td>Surgical Scissors - Sharp_Blunt</td>
 <td>Fine Science Tools</td>
 <td>14001-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Iris Scissors - ToughCut Straight 11.5 cm</td>
 <td>Fine Science Tools</td>
 <td>14058-11</td>
 <td></td>
 </tr>
 <tr>
 <td>Student Vannas Spring Scissors </td>
 <td>Fine Science Tools</td>
 <td>91500-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Vannas Spring Scissors - 3 mm Blades</td>
 <td>Fine Science Tools</td>
 <td>15000-00</td>
 <td></td>
 </tr>
 <tr>
 <td>Student Tissue Forceps - 1x2 Teeth 12 cm</td>
 <td>Fine Science Tools</td>
 <td>91121-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Dumont SS-45 Forceps - Inox Medical</td>
 <td>Fine Science Tools</td>
 <td>11203-25</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine Clip Applicator with Lock</td>
 <td>Fine Science Tools</td>
 <td>18056-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine 6 mm x 1 mm</td>
 <td>Fine Science Tools</td>
 <td>18055-03</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine 4 mm x 0.75 mm</td>
 <td>Fine Science Tools</td>
 <td>18055-04</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent</td>
 </tr>
 <tr>
 <td>Isoflurane (e.g. Forene 100% v/v)</td>
 <td>Abott</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>cutane antiseptic (e.g. Octeniderm) </td>
 <td>Sch&uuml;lke</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Povidone Iodine (e.g. Betaisodona)</td>
 <td>Mundipharma</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>ophthalmic ointment (e.g. Bepanthen)</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprenorphin (e.g. Temgesic)</td>
 <td>RB Pharmaceuticals</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>HTK perfusion solution (e.g. CUSTODIOL HTK)</td>
 <td>Dr. Franz K&ouml;hler Chemie</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Mersilene 0</td>
 <td>Ethicon</td>
 <td>EH6665E</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Mersilene 4-0</td>
 <td>Ethicon</td>
 <td>EH6732H</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Prolene 6-0</td>
 <td>Ethicon</td>
 <td>8697H</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Ethilon 9-0</td>
 <td>Ethicon</td>
 <td>2809G</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical silk 5-0</td>
 <td>V&ouml;mel</td>
 <td>14739</td>
 <td></td>
 </tr>
 <tr>
 <td>Canula (e.g. Microlance 3, 27G &frac34;)</td>
 <td>BD</td>
 <td>302200</td>
 <td></td>
 </tr>
 </tbody>
</table>

non-invasive imaging of acute allograft rejection after rat renal transplantation using 18f-fdg pet

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.

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...

Watch this Scientific Journal Video about Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET at JoVE.com

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Research

Education

JoVE Journal

JoVE Core

Biology

Medicine

Human Biology

Osmoregulation and Excretion

SCIENTISTS IN ACTION

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...