An Adenoviral Vector Encoding Full-Length Dectin-1 Promotes Aspergillus-Induced Innate Immune Response in Macrophages

• DOI: 10.1007/s00408-015-9740-8
• Publication Date: 2015-05-05T07:25:16Z
• Authors: Di Xia; Wen-Kui Sun; Ming-Ming Tan; Yuan Ding; Zhi-Cheng Liu; Pei Li; Qian Qian; Xin Su; Yi Shi
• Keywords: 0301 basic medicine; Tumor Necrosis Factor-alpha; Aspergillus fumigatus; Genetic Vectors; Interleukin-1beta; Gene Expression; Receptors, Cell Surface; Spores, Fungal; Transfection; Immunity, Innate; Adenoviridae; Interleukin-10; Up-Regulation; 3. Good health; Mice; 03 medical and health sciences; Phagocytosis; Macrophages, Alveolar; Animals; Lectins, C-Type; RNA, Messenger

The incidence of invasive pulmonary aspergillosis (IPA) has increased significantly over the last two decades. Alveolar macrophages (AMs) represent the first line of pulmonary host response to Aspergillus conidia. Recognition of conidia by AMs involves Dectin-1 (CLEC7A), which is a conserved structure to combine β-glucans. The deficiency of Dectin-1 results in impaired fungal killing and uncontrolled growth of Aspergillus fumigatus. Thus, we hypothesized that high expression of Dectin-1 would enhance the host recognition and fungal killing.We set out to develop an adenoviral vector encoding full-length Dectin-1 (Ad-Dectin-1-EGFP) and then transfect it to MH-S cells. Transfect cell model was verified by using real-time RT-PCR, Western blot, flow cytometric, and confocal microscopic assays. And also, the function of Dectin-1 was explored by measuring cytokine release and killing ability during the course of A. fumigatus infection.We constructed a recombinant adenovirus which could upregulate the expression of Dectin-1 and verified that Dectin-1 was expressed on cell membrane. The function of Dectin-1 was also demonstrated by its ability in promoting the production of cytokines and increasing the killing ability during the course of A. fumigatus infection.An adenoviral vector was successfully applied to the production of a recombinant adenovirus encoding full-length Dectin-1, and also, its function in Aspergillus-induced innate immune response was demonstrated.