J Korean Soc Transplant.  2013 Sep;27(3):114-120. 10.4285/jkstn.2013.27.3.114.

Validation of Tissue Microarrays for the Study of Immunosuppressive Agent-induced Nephrotoxicity

• Lim BJ
• Kim JH
• Jeong HJ

• KMID: 1495868
• DOI: http://doi.org/10.4285/jkstn.2013.27.3.114

Abstract

BACKGROUND
Tissue microarray analysis (TMA) is a high-throughput method for histologic evaluation, immunohistochemistry, and in situ hybridization using paraffin embedded tissue. Despite its high efficiency as an experimental tool, TMA is limited because it only contains a very small tissue fragment from each case. Therefore, the purpose of this study was to evaluate the validity of TMA in a study of nephrotoxicity caused by immunosuppressants.
METHODS
Male Sprague-Dawley rats were treated with vehicle (n=16), cyclosporine (n=23), and cyclosporine plus losartan (n=13) for a maximum of 7 weeks. After animal sacrifice, renal tissues were embedded in paraffin and processed into slides for microscopic examination using conventional methods and the TMA technique. Acute tubular injury, vascular hyaline change, and interstitial fibrosis were scored in both conventional and TMA slides. The number of interstitial macrophages was counted after ED-1 immunohistochemistry and the results also compared between conventional and TMA slides.
RESULTS
The degree of acute tubular injury and interstitial fibrosis showed a significant agreement between conventional and TMA methods (kappa value, 0.79 and 1.00, respectively). The number of interstitial macrophages counted in conventional and TMA slides showed a significant correlation as well (r=0.934, P<0.001). However, the degree of vascular hyaline changes showed less agreement between conventional and TMA methods (kappa value, 0.40).
CONCLUSIONS
TMA is a useful and reliable method for the study of nephrotoxicity induced by immunosuppressive agents. TMA also reflects the findings of conventional methods, especially for acute and chronic tubular and interstitial changes.

Keyword

Tissue microarray analysis; Acute kidney injury; Tubulointerstitial fibrosis; Cyclosporine

MeSH Terms

Acute Kidney Injury
Animals
Cyclosporine
Fibrosis
Humans
Hyalin
Immunohistochemistry
Immunosuppressive Agents
In Situ Hybridization
Losartan
Macrophages
Male
Paraffin
Rats, Sprague-Dawley
Tissue Array Analysis
Vascular System Injuries
Cyclosporine
Immunosuppressive Agents
Losartan
Paraffin

Figure

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  Fig. 1. A sample of tissue microarray (TMA). (A) 1:1 photography of a paraffin block and (B) a PAS-stained slide. Each TMA contained upto 30 tissue cores. Tissue cores were 3 mm in diameter and obtained from corticomedullary junction. Representative photography of cores (C, ×40; D, ×200).

• 

  Fig. 2. ED-1 positive cell infiltration observed in conventional and tissue microarray analysis (TMA) slides. (A) In conventional slides, the number of ED-1 positive cells in interstitium was markedly increased after cyclosporine treatment, and remained in low level after cyclosporine and losartan combination treatment. (B) The result obtained from TMA slides showed same tendency in each group. Abbreviations: HPF, high power field; CsA, cyclosporine; LSRT, losartan.

• 

  Fig. 3. Correlation of ED-1 positive cell counts obtained from conventional and tissue microarray analysis (TMA) slides. The number of ED-1 positive cells counted in conventional and TMA slides showed a significant correlation in each case (γ=0.934, P＜0.001). Abbreviation: HPF, high power field.

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