Original Article

Use of laser microdissection in the analysis of renal-infiltrating T cells in MRL/ lpr mice

Authors

Yingge Wang¹, Satoshi Ito¹, Yusuke Chino¹, Keiichi Iwanami¹, Takanori Yasukochi¹, Daisuke Goto¹, Isao Matsumoto¹, Taichi Hayashi¹, Kazuhiko Uchida² and Takayuki Sumida¹

1. Division of Clinical Immunology, Major of Advanced Biomedical Applications, Graduate School of Comprehensive Human Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba Ibaraki, 305-8575, Japan
2. Molecular and Biological Oncology, Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Japan

Received:

25 January 2008

Accepted:

12 March 2008

Published online:

2 May 2008

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Abstract

To clarify the role of T cells in the kidneys of MRL/MpJ-lpr (MRL/lpr) mice, cytokine mRNA expression was analyzed, and tissue localization of T cells was examined by immunohistochemistry. Cells infiltrating the glomeruli, glomerular circumference, and perivascular areas in ten female MRL/lpr mice were captured by laser microdissection (LMD). Nested reverse transcription polymerase chain reaction (RT-PCR) of samples was performed with primers specific for β-actin, T-cell receptor β chain (TCR-Cβ), Thy-1, B220, CD4, CD8, interleukin (IL)-2, IL-4, IL-10, IL-13, IL-17, and interferon (IFN)-γ. Frozen sections of lesions were also stained immunohistochemically. B220, MAC-1, Thy-1, CD4, and CD8 staining was observed in glomeruli and perivascular areas, especially in glomerular circumference areas. T cells infiltrating the glomeruli, glomerular circumference areas, and perivascular areas produce INF-γ, IL-13, and IL-17 predominately. IL-10 positivity was identified in 60% of perivascular T cells but not in a substantial number of glomerular or periglomerular T cells. The results of our study suggest that the pathogenesis of renal lesions in MRL/lpr mice is complex and not due simply to the Th1 and Th2 balance. These findings also support the concept of different molecular mechanisms for glomerulonephritis and vasculitis in these mice.

Key words

Glomerulonephritis - Laser microdissection (LMD) - MRL/lpr mice - T cells - Vasculitis