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Archive > Volume 24 Issue 7 > 2017,24(7):721-726. DOI:10.3872/j.issn.1007-385X.2017.07.005 Prev Next
Monographic Report
Effects of NKG2D ligand RAE1ε on the function of 4T1 mammary cancer cells-derived MDSC
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Abstract:
Objective: To explore effect of BaF3 primary B lymphocyte that expresses retinoic acid early transcript 1? (RAE1?), ligand of mouse NKG2D, on function of myeloid-derived suppressor cell (MDSC) originated from mouse. Methods: Based on mouse primary B lymphocyte BaF3 line, a BaF3-RAE1? cell that expressed RAE1? and a BaF3-mock control cell with empty plasmid were structured. CD11b + Gr-1 + MDSC was produced through introduc- tion of 4T1 tumor model in situ. Spleen MDSC was respectively co-cultured with BaF3-RAE1? cell and BaF3-mock cell that acted as stimulating cells. Flow cytometry assay was used to detect expressions of CD40, CD80, F4/80 , CD11c and content of reactive oxygen species (ROS) in the MDSC. ELISA assay was used to test contents of IL-10, IL-4 ans IFN-γ in supernatant of the co-culture. Concentration of nitric oxide (NO) in the supernatant was examed by Griess assay. Magnetic beads were used to separate the MDSC in the co-culture system, activity of arginase was detected after splitting. In addition, the separated MDSC co-culture with splenocyte activated by anti-CD3/CD28 an-tibody. Proliferation of the activated CD3 + CD8 + T cell was examed by CFSC assay. Results: The MDSC of mouse spleen derived from 4T1 in situ tumor model was successfully obtained. Comparing with the BaF3-mock cell, effect- ing of the BaF3-RAE1? cell stimulation on amounts of IL-4, IFN-γ, IL-10 and NO secreted by the MDSC was not significant (P>0.05), and on amounts of CD40, CD80, F4/80 ,CD11c and ROS expressed by the MDSC also not sig- nificant (P>0.05). Stimulation of the BaF3-RAE1? cell did more increase activity of arginase in the MDSC than that of the BaF3-mock cell obviously (156.166±1.209 vs 110.135±7.356, P<0.01), and evidently enhanced inhibitory effect of the MDSC on proliferation of CD8 + T cell. Conclusion: RAE1? could enhance inhibitory function in vitro of the MDSC derived from 4T1 mouse with tumor.
Keywords:
retinoic acid early transcript 1ε ; myeloid-derived suppressor cell ; arginase ; 4T1 cell of breast cancer
Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81373130, No.81001308) ，the Natural Science Foundation of Jiangsu Province (No. BK2010315), and the Science and Technology Innovation Fund for Undergraduates of Yangzhou University
