Phytohaemagglutinin stimulates the proliferation of peripheral blood mononuclear cells and expression of secretory cytokines / 中国组织工程研究

ABSTRACT

BACKGROUND:Phytohemagglutinin (PHA) can stimulate the peripheral blood mononuclear cells (PBMCs) into cellcycle, and cause their immune activation, which is a common immune proliferation model. However, the role of non-PBMC ingredient of peripheral blood is unclear, as wel as the expression of endothelial cells related cytokines. OBJECTIVE:To study the effect of whole blood culture and PBMCs alone culture with PHA on the PBMC proliferation and apoptosis, expression of inflammatory cytokine and endothelial cellsecreted cytokine markers. METHODS:Morphological changes of PBMCs separated from normal karyotype human peripheral blood individual y cultured with or without PHA were observed. The PBMCs were col ected by whole blood culture or PBMC separated culture. mRNA was extracted for the fluorescence quantitative RT-PCR, which was applied to detect the cellproliferation, apoptosis, and expression of inflammatory cytokine and endothelial cellsecreted cytokines. The statistic analysis was used for the significance explication. RESULTS AND CONCLUSION:PBMCs alone cultured ere different from those undergoing whole blood culture. The PHA could up-regulate the gene expression of Ki67, proliferating cellnuclear antigen, Caspase 3, interferon-γ, tumor necrosis factor-βand interleukin-6, but down-regulate Protein C. This indicted that PHA could promote the proliferation and apoptosis of PBMCs and up-regulate the expression of inflammatory cytokines, but down-regulate the expression of endothelial cells secreted coagulation cytokines.