Name: mouse kidney transplantation: models of allograft rejection
Uploaded: 2014-10-11T15:00:00
Description: Watch this Scientific Journal Video about Mouse Kidney Transplantation: Models of Allograft Rejection at JoVE.com

Funding from Kidney Research UK, The Royal College of Surgeons of Edinburgh and The European Society of Organ Transplantation supported this study.

Appropriate national and local institutional ethics should be in place before performing animal experiments. Specifically in the United Kingdom the following experiments were undertaken under the Animals (Scientific Procedures) Act 1986. Where two microsurgeons are available to operate simultaneously the donor surgeon should perform steps 1.1 to 1.16 then 3.1 to 3.5, whilst the recipient surgeon performs 2.1 to 2.8. For a single operator the steps can be followed sequentially.
1. Donor Preparati...

<ol>
	<li>Guild, W. R., Harrison, J. H., Merrill, J. P., Murray, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+homotransplantation+of+the+kidney+in+an+identical+twin.">Successful homotransplantation of the kidney in an identical twin.</a> Trans. Am. Clin. Climatol Assoc. 67, 167-173 (1955).</li><li>Wolfe, R. A., 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=Comparison+of+mortality+in+all+patients+on+dialysis,+patients+on+dialysis+awaiting+transplantation,+and+recipients+of+a+first+cadaveric+transplant.">Comparison of mortality in all patients on dialysis, patients on dialysis awaiting transplantation, and recipients of a first cadaveric transplant.</a> N. Engl. J. Med. 341, 1725-1730 (1999).</li><li>Nankivell, B. J., Alexander, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rejection of the Kidney Allograft. N. Engl. J. Med. 363, 1451-1462 (2010).</li><li>Tse, G. H., Hughes, J., Marson, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+review+of+mouse+kidney+transplantation.">Systematic review of mouse kidney transplantation.</a> Transplant International. 26, 1149-1160 (2013).</li><li>Skoskiewicz, M., Chase, C., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplants+between+mice+of+graded+immunogenetic+diversity.+Transplant.">Kidney transplants between mice of graded immunogenetic diversity. Transplant.</a> Proc. 5, 721-725 (1973).</li><li>Zhang, Z., 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=Pattern+of+liver,+kidney,+heart,+and+intestine+allograft+rejection+in+different+mouse+strain+combinations.">Pattern of liver, kidney, heart, and intestine allograft rejection in different mouse strain combinations.</a> Transplantation. 62, 1267-1272 (1996).</li><li>Qi, F., 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=Depletion+of+cells+of+monocyte+lineage+prevents+loss+of+renal+microvasculature+in+murine+kidney+transplantation.">Depletion of cells of monocyte lineage prevents loss of renal microvasculature in murine kidney transplantation.</a> Transplantation. 86, 1267-1274 (2008).</li><li>Dang, Z., Mackinnon, A., Marson, L. P., Sethi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tubular+atrophy+and+interstitial+fibrosis+after+renal+transplantation+is+dependent+on+galectin-3.">Tubular atrophy and interstitial fibrosis after renal transplantation is dependent on galectin-3.</a> Transplantation. 93, 477-484 (2012).</li><li>Jabs, W. J., 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=Heterogeneity+in+the+Evolution+and+Mechanisms+of+the+Lesions+of+Kidney+Allograft+Rejection+in+Mice.">Heterogeneity in the Evolution and Mechanisms of the Lesions of Kidney Allograft Rejection in Mice.</a> Am. J. Transplant. 3, 1501-1509 (2003).</li><li>Lin, T., 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=Deficiency+of+C4+from+Donor+or+Recipient+Mouse+Fails+to+Prevent+Renal+Allograft+Rejection.">Deficiency of C4 from Donor or Recipient Mouse Fails to Prevent Renal Allograft Rejection.</a> Am. J. Pathol. 168, 1241-1248 (2006).</li></ol>

The most well described manner to perform the arterial anastomosis is to use the distal aorta of the donor, with the renal artery in continuation, in an end-to-side manner to the recipient aorta. We describe the use of an aortic patch, similar to the &#8216;Carrell patch&#8217; mirroring that performed in human kidney transplantation which we believe to be more convenient. Though reports in the literature of donor and recipient operative time are sparse we believe that utilizing an aortic patch to recipient aorta instead...

Successful renal transplantation for the treatment of renal failure was first described in 1955 between monozygotic twins1, since then it has become a revolutionary treatment for patients with end-stage renal failure throughout the world, offering both improvement in length and quality of life2. However long term graft survival has been hampered by a multitude of pathological processes resulting in chronic allograft damage3.
Rejection of the transplanted kidney in humans remains a major cause of morbidity, despite significant improvements in immunosupporessive regimens. The aim of developing a mouse model of renal transplantation is to closely replicate the process and pathology found in human renal transplantation4. Skoskiewicz et al. first described the mouse model of renal transplantation in 19735. Although advanced microsurgical skills are required, it is a valuable tool for several reasons: the mouse genome has been well characterized and there is a great variety of experimental methods and techniques available for mouse studies.
Many groups using the mouse model of renal transplantation have used the transplanted kidney as a life-supporting organ, however in other studies and in our described methodology one of the recipient mouse&#8217;s native kidneys is left in situ for the duration of the experiment4. The benefit is that the mouse undergoes a single anesthesia and operation thereby reducing the morbidity to the mouse and the risk of death from a second procedure. Additionally the mouse does not suffer from the adverse effects of gradual renal failure.
Although models of allogeneic rejection exist in other organs such as the heart and skin, these are not always directly relevant to renal transplantation. There is evidence that these models elicit different modes and dynamics of rejection, for instance the time course of rejection in cardiac allograft and renal allograft differs significantly in certain strain combinations6. We have described acute renal allograft rejection patterns in BALB/c donors into non-transgenic FVB/nj mice, this model showed cellular mediated injury with accumulation of T cells and macrophages7. Alternatively we have also described a model of chronic allograft damage that exhibits interstitial fibrosis and tubular atrophy, this results from transplanting a kidney from C57BL/6BM12 donors into C57BL/6 recipients, as these mice are characterized by a single MHC class II loci mis-match8.
Multiple aspects of transplantation have been studied using the mouse model of renal transplantation including acute rejection, cellular and humoral rejection, ischemia reperfusion injury, and trialing novel therapeutic agents. We have modified the surgical technique to reduce operating time and improve the ease of surgery. Particularly we have described simultaneous donor and recipient preparation and a simplified vascular anastomosis technique by utilizing a continuous aortic patch anastomosis. This video and manuscript will provide key points to aid in the establishment of this technique.

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			<td>Fine Science Tools</td>
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			<td>Curved Castoviejo scissors</td>
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			<td>Spring Scissors &#8211; straight</td>
			<td>Fine Science Tools</td>
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			<td>Regular bevel needle, 1/2 inch, 30G</td>
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			<td>Bection, Dickinson and Company</td>
			<td>324827</td>
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			<td>Insulin needle 0.3ml, 30G</td>
			<td>Bection, Dickinson and Company</td>
			<td>324826</td>
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			<td>1 ml syringe slip tip</td>
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			<td>(Lacri-Lube) White soft paraffin 57.3%, mineral oil 42.5% and lanolin alcohols 0.2%</td>
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			<td>PL 04509/0041</td>
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			<td>(Vetalar V) Ketamine hydrochloride</td>
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			<td>University of Wisconsin Solution</td>
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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 replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Renal allograft rejection can be assessed by histological analysis of methacarn-fixed paraffin-embedded tissue sections of the transplanted kidney (Figure 2). Isograft transplantation of kidneys between syngeneic mice results in renal ischemic reperfusion injury, however by 4 weeks the tubules have recovered and are histologically comparable to native kidneys. Acute rejection can be modeled by C57BL/6 kidney transplantation into BALB/c recipients, within 1 week there is diffuse mononuclear cell infiltrat...

Watch this Scientific Journal Video about Mouse Kidney Transplantation: Models of Allograft Rejection at JoVE.com

Mouse Kidney Transplantation: Models of Allograft Rejection

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

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

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