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Effects of genetically modified pro-B lymphocytes on the expression of functional molecules in interferon-producing killer dendritic cells
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Abstract:
Objective: To study the effects of murine bone marrow-derived pro-B (BaF3) cells overexpressing membrane-bound form of IL-15 (mbIL-15) and retinoic acid early transcript 1ε (RAE1ε), either each alone or two in combination, on the expression of functional molecules in murine interferon producing killer dendritic cells (IKDCs) in vitro . Methods: Clones of BaF3 cells overexpressing mbIL-15 , RAE1ε or both mbIL-15 and RAE1ε were established and inactivated by mitomycin. Mouse bone marrow mononuclear cells were isolated and induced into mature DCs by GM-CSF and IL-4. Mature DCs were co-cultured with the three mitomycin inactivated-BaF3 derivatives respectively. At 24 and 72 hours after co-culture, the percentage of CD11clow B220+NK1.1+ IKDCs and levels of CD40 and CD80 in DCs and levels of CD40, CD80, CD86, NKG2D, MHC class Ⅱ molecule, CD107a and FasL in IKDCs were determined by flow cytometry. Results: BaF3 cells overexpressing both mbIL-15 and RAE were more effective than BaF3 cells overexpressing mbIL-15 or RAE alone to stimulate the DC to IKDC conversion (\[50.0±5.6\]% vs \[30.3±8.2\]%, \[36.0±4.6\]%, P <0.05). IKDCs co-cultured with BaF3 cells overexpressing mbIL-15 and RAE had higher levels of CD40 (180.1±282 vs 44.7±7.8, 36.0±3.1, P <0.01) and FasL ( P <0.05) as compared to IKDCs co-cultured with mbIL-15- or RAE-overexpressing BaF3 cells. IKDCs co-cultured with BaF3 cells overexpressing mbIL-15 and RAE had higher levels of CD80 ( P <0.05) as compared to IKDCs co-cultured with BaF3 cells or mbIL-15-overexpressing BaF3 cells. In contrast, co-culture with BaF3 derivatives showed no effects either on CD40 and CD80 expression in DCs ( P >0.05)or on CD86, MHC class Ⅱ molecule, NKG2D and CD107a expression in IKDCs. Conclusion: IL-15 combined with RAE1ε may promote the proliferation and CD40 and FasL expression in IKDCs.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81001308，No. 81373130，No. 81273214) ，the Natural Science Foundation of Jiangsu Province (No.BK2010315)，the Science and Technology Innovation Fundation for Undergraduates of the Educational Committee of Jiangsu Province, and the New Century Talents Project of Yangzhou University
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