Use of laser microdissection in the analysis of renal-infiltrating T cells in MRL/lpr mice.
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 beta-actin, T-cell receptor beta chain (TCR-Cbeta), Thy-1, B220, CD4, CD8, interleukin (IL)-2, IL-4, IL-10, IL-13, IL-17, and interferon (IFN)-gamma. 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-gamma, 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.
Wang
Yingge
Y
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.
Ito
Satoshi
S
Chino
Yusuke
Y
Iwanami
Keiichi
K
Yasukochi
Takanori
T
Goto
Daisuke
D
Matsumoto
Isao
I
Hayashi
Taichi
T
Uchida
Kazuhiko
K
Sumida
Takayuki
T
D000818:Animals; D016207:Cytokines / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D020869:Gene Expression Profiling; D053685:Laser Therapy; D008181:Lupus Nephritis / Q000276:immunology / Q000473:pathology*; D051379:Mice; D042282:Microdissection / Q000379:methods; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology*; D014657:Vasculitis / Q000473:pathology*
D016207:Cytokines