Journal List > J Korean Soc Transplant > v.27(3) > 1034397

J Korean Soc Transplant. 2013 Sep;27(3):114-120. Korean.
Published online September 26, 2013.  https://doi.org/10.4285/jkstn.2013.27.3.114
Copyright © 2013 The Korean Society for Transplantation
Validation of Tissue Microarrays for the Study of Immunosuppressive Agent-induced Nephrotoxicity
Beom Jin Lim, M.D.,1 Ji Hong Kim, M.D.,2 and Hyeon Joo Jeong, M.D.1
1Department of Pathology, Yonsei University College of Medicine, Seoul, Korea.
2Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea.

Corresponding author (Email: jeong10@yuhs.ac )
Received May 08, 2013; Accepted June 30, 2013.

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 (κ 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 (κ 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.

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

Figures


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).
Click for larger image


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.
Click for larger image


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.
Click for larger image

Tables


Table 1
Comparison of the severity of tubular injury between conventional and tissue microarray analysis slides
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Table 2
Comparison of the severity of vascular hyaline change between conventional and tissue microarray analysis slides
Click for larger image


Table 3
Comparison of the degree of interstitial fibrosis between conventional and tissue microarray analysis slides
Click for larger image

References
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3. Solez K, Colvin RB, Racusen LC, Haas M, Sis B, Mengel M, et al. Banff 07 classification of renal allograft pathology: updates and future directions. Am J Transplant 2008;8:753–760.
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