eISSN: 1644-4124
ISSN: 1426-3912
Central European Journal of Immunology
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vol. 39

Experimental immunology
Use of laser microdissection in the analysis of of renal infiltrating T cells in murine lupus

Zhentao Guo
Yingge Wang
Rongqian Li
Haifeng Huang
Rong Wang

(Centr Eur J Immunol 2014; 39 (3): 285-293)
Online publish date: 2014/10/14
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Objective: To clarify the role of T cells in kidney pathology of three widely used murine lupus models.

Material and methods: Cells infiltrating the glomeruli and perivascular areas in MRL/lpr (n = 10 female), NZB× NZW F1 (B/W F1) (n = 9 female), and BXSB (n = 10 male) mice were captured by laser microdissection (LMD). Samples were subjected to nested reverse transcription polymerase chain reaction (RT-PCR) with primers specific to β-actin, T-cell receptor β chain (TCR-Cβ), interleukin (IL)-10, IL-13, IL-17, and interferon- (IFN-). Frozen sections of lesions were also stained immunohistochemically for tissue and cellular characterization.

Results: T cells infiltrating the glomeruli and perivascular areas predominantly produced IFN-, IL-13, and IL-17 in MRL/lpr, B/W F1, and BXSB mice, with IL-17 expression in glomeruli of BXSB mice being significantly lower than that of MRL/lpr and B/W F1 mice. IL-10 was detected only in the perivascular areas of MRL/lpr and B/W F1 mice and not in glomeruli isolates. Immunohistochemical staining revealed positive for the expression of Thy-1, CD4, CD8, and B220 in glomeruli and perivascular areas from all three strains of mice.

Conclusions: Cytokine balance in murine SLE is complex and cannot be attributed simply to the balance between Th1 and Th2 cells. Th17 cells may play a critical role in disease pathology, possibly with greater contribution toward disease progression in MRL/lpr and B/W F1 mice than in BXSB mice. Furthermore, these findings lend support to the concept that different molecular mechanisms underlie glomerulonephritis as compared to vasculitis.

murine lupus, glomerulonephritis, vasculitis, laser microdissection, IL-17

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