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CRACC-CRACC interaction between macrophages and NK cells enhances the cytokine secretion in vitro.

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posted on 30.09.2013, 02:37 by Yangxi Li, Guoshuai Cao, Xiaodong Zheng, Jun Wang, Haiming Wei, Zhigang Tian, Rui Sun

A, Ana-1 cells pre-transfected with liposome encapsulated siNeg or siCRACC was subsequently stimulated with Poly I:C. The expression of CRACC mRNA was assayed by quantitative PCR at 3h time point post the Poly I:C stimulation. B, C & D, Ana-1 cells pre-transfected with nanoparticle encapsulated siNeg or siCRACC was cultured in absence or presence of NK cells, which were freshly isolated from mice pre-treated by Poly I:C. Then Ana-1 cells cultured with or without NK cells were stimulated with Poly I:C; and D-GalN was made as control. CRACC expression on Ana-1 cells was analyzed by flow cytometry (B) and the cytokine (TNF-α and IFN-γ) in the culture supernatant was assayed by ELISA (C, D) at 24h time point post the Poly I:C or D-GalN stimulation; Ana-1 cells were identified as F4/80+ cells. E, Ana-1 cells were cocultured with freshly isolated NK cells from mice pre-treated by Poly I:C; and Ana-1 cells alone or NK cells alone were made as control. The Ana-1-NK coculture was stimulated by Poly I:C in the presence of isotype control Ab (Rat IgG), CRACC mAb (anti-CRACC), or NKG2D mAb (anti-NKG2D), respectively. The IFN-γ in culture supernatant was assayed by ELISA at 24h time point post the Poly I:C stimulation. *P < 0.05; **P<0.01; ***P < 0.001.

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