ELMO1 gene is located on chromosome number seven in humans, and the gene is positioned on chromosome number thirteen in mice, this gene encoded ELMO1 protein which plays an important role in cells phagocytosis and migration, while misguided cell migration and phagocytosis will lead to inflammatory related diseases even metastatic cancer. Macrophages derived from mononuclear cells of bone marrow which exist all body tissues, and it is the most flexibility immune cells in the hematopoietic system. Macrophages are not only an important component of nonspecific immunity, but also the role of presenting antigen in specific immune responses. It has a variety of functions in maintaining the body’s development, homeeostasis, tissue repair and immunity response. In the non-specific immunity, macrophage can remove the invading pathogens, dead cells or cell debris by phagocytosis. In specific immune responses, it can uptake and present exogenous materials, through the secretion of cytokines and chemokines induce and modulate the immune response. Almost every human disease is related to macrophages, which are also used as the main treatment goal. Because the diversities of functions can be oriented enhance or inhibition, it can change the outcome of the disease. LPS stimulate macrophages through TLR4 receptors to adjust the expression of cytokines. ELMO1 gene is involved in cell migration and phagocytosis, and macrophages play an important role in body’s innate immunity and adaptive immunity. If ELMO1 can adjust phagocytosis and secretion in macrophage, and what is the molecular mechanism involved in? Firstly, our laboratory was successfully constructed the vector PLVX-sh RNA which interference ELMO1 expression and control plasmid, package lent virus and then infected the RAW264.7 cell lines, sort stable cell by flow cytometry after expanding cultured, observe the cell morphology in vivo.Using cell lines engulf beads labeled with fluorescent tags, FACS analysis phagocytosis efficiency. Then, we successfully established ELMO1 knockout mice, achieved primary macrophages from mouse peritoneal macrophages or induced from bone marrow cells, through the phagocytic index method detect the phagocytic efficiency of the wild type and knockout macrophage.We detect the inflammatory cytokines expression after LPS stimulation, as well as the activation of MAPKs.The results showed that: the phagocytosis is weaker after knockdown ELMO1 gene in RAW264.7 cells,in mouse primary cells, the expression of Dock2 decreased in knockout cells than wild-type cells,but phagocytic ability of cells are no significant change, which may be due to the elevated expression of ELMO2 which is another subtype of ELMO in knockout mice. With LPS stimulation macrophage, the secretion of TNF-α and IL-6 is increase in ELMO1 knockout macrophages. ELMO1 combine to Dock2 forming a complex. the phagocytosis in primary macrophages should be further explored. |