Kidney Week

Abstract: FR-PO094

TissueFAXS Analysis as an Experimental Tool for Assessing the Degree of Intrarenal Lymphocytes Infiltration

Session Information

• AKI Clinical: Predictors
  November 03, 2017 | Location: Hall H, Morial Convention Center
  Abstract Time: 10:00 AM - 10:00 AM

Category: Acute Kidney Injury

• 003 AKI: Clinical and Translational

Authors

• Kim, Minjung, Samsung medical center, Seoul, Korea (the Republic of)

Minjung Kim,

• Kim, Su Hyun, samsung medical center, Seoul, Korea (the Republic of)

Su Hyun Kim,

• Jeon, Junseok, Samsung medical center, Seoul, Korea (the Republic of)

Junseok Jeon,

• Lee, Jung eun, Samsung medical center, Seoul, Korea (the Republic of)

Jung eun Lee,

• Huh, Wooseong, Samsung medical center, Seoul, Korea (the Republic of)

Wooseong Huh,

• Kim, Yoon-Goo, Samsung medical center, Seoul, Korea (the Republic of)

Yoon-Goo Kim,

• Kim, Dae Joong, Samsung medical center, Seoul, Korea (the Republic of)

Dae Joong Kim,

• Oh, Ha Young, Samsung medical center, Seoul, Korea (the Republic of)

Ha Young Oh,

• Jang, Hye Ryoun, Samsung medical center, Seoul, Korea (the Republic of)

Hye Ryoun Jang,

Background

Intrarenal infiltration of immune cells is an established pathogenic mechanism in various inflammatory kidney diseases. Quantitative analyses of intrarenal immune cells, especially intrarenal lymphocytes, using flow cytometry analysis of kidney mononuclear cells (KMNCs) have been used as a precise and useful diagnostic method in a murine model of ischemic acute kidney injury (AKI). However, flow cytometry analysis of KMNCs is not feasible in patients because sufficient amount of renal tissue required for this method cannot be obtained with kidney biopsy. In this study, we investigated the diagnostic potential of tissueFAXS as a tool assessing the degree of intrarenal lymphocytes infiltration.

Methods

A total of 10 cynomolgus monkeys with various renal function were included. Kidney samples for flow cytometry analysis of KMNCs and immunohistochemistry were collected simultaneously. Flow cytometry analysis of KMNCs were performed using FACS verse. Immunohistochemistry of CD3, CD20, and CD45 on formalin-fixed kidney tissues were performed and followed by tissueFAXS analysis. The proportion of lymphocytes positive for each CD marker was acquired using different denominators as follows: kidney mononuclear cells isolated with percoll in flow cytometry vs. all nucleated cells stained with hematoxylin in tissueFAXS. Linear regression analysis was used to compare the association of CD3, CD20, and CD45 (+) lymphocytes evaluated with two different methods.

Results

Flow cytometry and tissueFAXS analyses showed positive correlation with CD45 (+) lymphocytes (R² =0.5378, P=0.0245). There was a positive correlation between the proportion of both CD3 (+) T cells (R² =0.8120, P=0.0002) and CD20 (+) B cells (R² =0.4587, P=0.0650). The ratio of CD3 (+) T cells and CD20 (+) B cells measured with both methods also showed positive correlation (R² =0.5144, P=0.0296).

Conclusion

Our study showed significant positive correlation of intrarenal CD3, CD20, and CD45 (+) lymphocytes measured with flow cytometry and tissueFAXS in cynomolgus monkeys. These results suggest that immunohistochemistry followed by tissueFAXS analysis may be used as a diagnostic tool performing semiquantitative evaluation of intrarenal lymphocytes in patients.